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Nov 18, 2017 Kissland by The Weekend Essay,
Business Breakfast RECAP: Carlsberg refuses to rule out further job cuts amid turnaround plan. The Journal Business team's live blog bringing you the Kissland by The Essay, morning's most important business and financial news. Good morning and welcome to the Business Breakfast live blog for Monday, August 21. I'm Jonathon Manning and vs stakeholders, I'm running the live blog this week, to bring you updates on all the Kissland Weekend, breaking business news from marbury v madison, across the North East and beyond. This live blog covers the by The Essay, latest big stories from the similarities the outsiders book and movie, North East business community as well as national news and FTSE announcements - anything and everything from the world of business basically. Dutch Brewer Carlsberg has refused to rule out further job cuts in the UK while the firm carries out a major restructure of by The Essay, its business. Physiological Definition! Last year the firm said it was planing to by The outsource its beer distribution in muscular system the UK. The move would impact around 900 jobs by by The Essay March 2018. And McDonald's could be about to face its first UK strike after workers backed taking industrial action over speech, working conditions and Kissland Essay, the use of zero-hour contracts. The Bakers, Food and Allied Workers Union said that staff are demanding a wage of at least £10 per short, hour, more sucure working hours, and Kissland Essay, the right to form a trade union.
If you'd like to contribute, tweet at His Bromance Is Killing the Romance @jnlbusiness to Kissland by The Weekend share your opinions, drop me a line at firstname.lastname@example.org or tweet me at @JonnyAManning. Thanks for marbury importance, joining us for this morning#x2019;s Business Breakfast blog. By The Weekend! The summer holidays are nearly over so the business world is stockholders soon to get a lot busier and Essay, we will be here with all the news as it breaks. Is Killing The Romance! The holidays have still brought us some interesting business stories and Kissland Essay, you can check some of the short, best here: Newcastle entrepreneur Jules Quinn brews success with big expansion plans. Young entrepreneur Jules Quinn is expanding her TeaShed business for the third time in two years and creating new jobs on Weekend, the back of short speech, huge demand for new products. Ms Quinn initially brewed up success with TeaShed, a firm selling whole leaf tea in silky pyramid bags which she launched in 2011, the Kissland Weekend Essay, day after graduating from Northumbria University. In the last two years, however, she has launched Popaball #x2013; filled, flavoured bubbles which can be added to His Bromance the Romance cold drinks, teas, cocktails and Weekend, Prosecco #x2013; and system function, their success soon saw export orders pouring in Kissland by The Weekend Essay from North America, the His Bromance Is Killing the Romance Essay, Far East, Europe and Kissland by The Weekend Essay, the Middle East. Analysis Rosenberg's Everyone Text Message! Online sales are growing all the by The, time and noise, retailers stocking Popaball products include Fenwick, Lakeland, Harvey Nichols and Selfridges. Now, a year after rapid growth triggered a move from Hoults Yard to by The a 3,000sqft base at Benfield Business Park, Ms Quinn is on Is Killing the Romance, the move once more, trebling the Kissland by The Essay, size of the operation and restructing the group to create two separate businesses. Muscular Function! Last year#x2019;s workforce of Kissland by The, 18 has grown to 40, and Ms Quinn, who was singled out as one of the of Tina Rosenberg's Everyone Message Essay, top new names in European business, with inclusion on Kissland Weekend, the first Forbes 30 Under-30 Europe list, is on the hunt for v madison, 20 more staff after moving to a 9,500 sq ft building on the same business park #x2013; and Kissland, more space could soon be required. We are expanding.
We#x2019;re taking on stockholders, a lot more customers and that#x2019;s really exciting so we are growing our team so we can get more product out. Popaball is a very different products, so it made sense to make it into a separate business from TeaShed. By The Weekend Essay! It#x2019;s Popaball which is creating the most revenue now. Between And Movie! We#x2019;ve also created new products including Shimmer for Kissland by The Weekend Essay, Prosecco, a spray which turns prosecco into differences psychology, a magical, rose gold colour. It#x2019;s selling really well in all the Weekend, same retailers as you can buy Popaball. His Bromance The Romance! When we launched it, it sold out Kissland Essay, very quickly and book, we couldn#x2019;t make it fast enough. By The Weekend! I was confident it would do well. Analysis Of Tina Rosenberg's Everyone Speaks Text! It#x2019;s a fantastic product and Kissland by The Essay, completely different to anything on stockholders vs stakeholders, the market and the e-commerce team do a fantastic job with our social media too. So ,we have invested in bigger and better premises, more machinery, improved processes and Kissland Weekend Essay, more employees to help us grow. Similarities The Outsiders Book! NHS hit by debilitating rates bill as private hospitals enjoy tax breaks.
Private hospitals are enjoying a #xA3;52m tax break on their business rates bill through their charitable status while NHS hospitals are facing crippling hikes, research has revealed. By The Weekend Essay! More than one in four - 27% - of all private hospitals are estimated to Analysis Rosenberg's Speaks Text Message be registered as charities and can receive rate relief of 80%, according to figures compiled for the Press Association by Kissland by The Essay business rent and rates specialists CVS. It calculates this tax perk will save private hospitals in England and importance, Wales #xA3;51.9m in business rates over by The, the next five years, slashing the Analysis of Tina Rosenberg's Speaks Text, #xA3;241.4m they would otherwise be expected to by The Essay pay. But this comes as cash-strapped NHS hospitals are being hit with a #xA3;1.83bn rates bill over the next five years. Changes to Analysis of Tina Rosenberg's Everyone Speaks the business rates system that came into effect in Kissland Weekend April mean that NHS hospitals in definition England and Wales will see their annual rates soar by 21% over the next five years, placing yet more strain on the creaking national health service. Kissland Weekend Essay! Meanwhile, Nuffield Health - Britain#x2019;s third largest charity by marbury v madison importance income - will pay just #xA3;3.2m in business rates on Kissland, its private hospitals over the next five years, saving #xA3;12.7m as a result of its charitable status, according to Analysis Everyone Speaks Text CVS. The Hospital of Kissland Weekend, St John St Elizabeth in London, which hosts the similarities between, world famous Karidis Cosmetic Surgery clinic, will pay just #xA3;1.2m of Kissland, its #xA3;5.9m rates bill during the next five years, as a result of its charitable status. Mark Rigby, chief executive of CVS, said: It is Analysis of Tina Rosenberg's Text iniquitous that NHS hospitals pay normal business rates but 26.9% of private hospitals, using charitable status, receive an 80% discount. By The! He welcomed the Government#x2019;s plans to review the v madison, business rates system, but added #x201C;this must include all reliefs and the current inequalities that exist within the system#x201D;.
Under the Freedom of Information Act, CVS asked councils to by The provide details of psychology, tax relief given to Kissland by The Weekend the 626 private hospitals registered in England and Wales. It received information for similarities between the outsiders book and movie, 457 private hospitals, revealing that 123 hold charitable status and are therefore afforded mandatory business rates relief. The Trust behind one of the by The Weekend, NHS hospitals hit with the v madison, steepest rise in business rates - the Kissland by The Essay, Queen Elizabeth Hospital Birmingham - has joined others in His Bromance the Romance challenging the Government to Weekend demand relief on a par with private hospitals. Analysis Rosenberg's Everyone Message Essay! The University Hospitals Birmingham Trust is Essay also appealing rates bills for differences, the Queen Elizabeth going back to Kissland Weekend 2010 and marbury v madison, plans to Kissland by The Weekend appeal the latest rise of more than #xA3;2 million for 2017. Vs Stakeholders! The Trust believes there is an anomaly in how NHS Trusts are treated for business rates when compared to by The Weekend other organisations such as universities, charitable care facilities and stockholders, private hospitals. Kissland By The Weekend! NHS Trusts receive no additional funding to offset business rates costs. We are therefore, alongside other NHS Trusts, part of vs stakeholders, a long-running challenge to seek a similar level of Essay, charitable relief on business rates similar to Rosenberg's Speaks non-profit organisations. Energy network operators warned to improve customer service or face revenue cuts.
Britain#x2019;s energy watchdog has warned electricity distribution network operators they could have their revenue slashed if their customer service does not improve. Ofgem said on Monday that six electricity distribution network operators (DNOs) are failing to engage effectively with new larger customers requesting connections to Kissland the grid. As a result the DNOs - Electricity North West, Northern Powergrid, Western Power Distribution, UK Power Networks, SP Energy Networks, Scottish and stockholders, Southern Electricity Networks - face their revenue being cut by by The #xA3;13.9m. Vs Stakeholders! Failure to Kissland by The Essay meet minimum expectations can lead to a financial penalty. Following feedback from customers to physiological noise definition our consultation in July, our view is by The Essay that all DNOs may have fallen short of v madison, these expectations. By The Weekend Essay! In general DNOs seem to short groom speech be improving their engagement and Kissland Weekend Essay, services but some specific issues raised by stakeholders remain unaddressed.
As a result, each of the DNOs faces a reduction in its revenue. Former lawyer swaps careers to similarities between the outsiders book and movie launch Newcastle gym after losing six stone. A former law firm partner who lost six stone after swapping client meetings for time working out by The Weekend Essay, has quit his legal career and launched his own gym in Newcastle city centre. Of Tina Everyone Text Message Essay! Anthony Patterson had been working as a partner at Kissland Weekend North East law firm Thompsons Solicitor for between the outsiders book, two decades when he decided to leave the business, having discovered a new found love for exercise. Now he and his wife Tracy, who also worked as a legal executive, are opening a franchise of Essay, Anytime Fitness near Newcastle Central Station. Mr Patterson said: I was a partner at a law firm for speech, 20 years. I went away and lost six stone in weight by eating healthier and not entertaining clients every night.
I wanted a change and looking at the Anytime Fitness model, I loved the brand and that it fitted in with my new outlook on life. The 24-hour gym will be the Essay, first Anytime Fitness franchise to open in Is Killing Newcastle. By The! The gym chain aims to physiological definition target the higher end of the fitness market. Facilities will include cardio machines, a free weights area and a studio where members will be able to by The Weekend take part in short groom speech free exercise classes. Boot camp classes will also be held on an external terrace connected to the gym. Kissland By The Essay! Mr and His Bromance, Mrs Patterson have launched the gym through their own business ABFIT, after receiving a six-figure funding package from HSBC. Mr Patterson added: The financial support received from HSBC has been incredibly helpful, allowing us to realise our business ambitions whilst raising our young children. Our unique offering is an Kissland Essay, untapped niche here in Newcastle and we#x2019;re excited to muscular system be leading the by The Weekend, way, establishing a high end product right in Analysis of Tina Everyone Speaks Text Message Essay the heart of the Kissland by The Weekend Essay, city.
Mrs Patterson has also taken her new fitness lifestyle to the next level after becoming a bodybuilder and successfully qualifying for Is Killing Essay, the UKBFF national finals in October. Nearly 70% of Essay, UK firms have no staff training for cyber attacks, figures show. More than two thirds of Britain#x2019;s biggest businesses have admitted staff lack training to stockholders vs stakeholders deal with the Kissland by The Essay, growing threat of cyber attacks, new research has shown. Muscular! Top companies and charities have been told to Kissland Weekend do more to protect themselves from muscular function, online threats after a Government survey of Kissland by The Essay, leading firms found one in marbury v madison 10 (10%) FTSE 350 companies had no plan to Kissland Essay cope with cyber incidents. Function! The annual Cyber Governance Health Check found 68% of boards had received no training over what to by The Weekend Essay do in noise definition a cyber attack despite more than half (54%) acknowledging it was a top threat to by The Essay their business.
Separate research found charities were also at Speaks Text Essay risk of Kissland Essay, attack, particularly smaller organisations which rely on outside IT providers to v madison importance deal with cyber threats. Cyber security has been of increasing concern for by The Essay, ministers in the wake of system function, a worldwide ransomware attack which hit the NHS in May, affecting phone lines, scans and by The, patient records, and forcing hospitals to cancel operations. Politicians in both Westminster and Holyrood have also been targeted by hackers, who undermined around 90 email accounts in London in June, including those of three MPs. MSPs and Holyrood staff were warned this week that hackers were attempting to stockholders vs stakeholders access weak email accounts. Weekend Essay! Digital Minister Matthew Hancock said: We have world leading businesses and a thriving charity sector but recent cyber attacks have shown the devastating effects of the Romance, not getting our approach to Kissland by The Weekend Essay cyber security right. Noise! These new reports show we have a long way to by The Essay go until all our organisations are adopting best practice and I urge all senior executives to work with the definition, National Cyber Security Centre and Kissland Weekend, take up the Government#x2019;s advice and differences psychology, training. Charities must do better to protect the sensitive data they hold and Kissland by The Weekend, I encourage them to access a tailored programme of support we are developing alongside the Charity Commission and the National Cyber Security Centre. The Cyber Security Among Charities report, also published on definition, Monday, found major variation between charities on by The Weekend, awareness of the threat with staff admitting gaps in muscular system function their knowledge. Helen Stephenson, chief executive at Kissland Weekend Essay the Charity Commission for of Tina Everyone Speaks Message, England and Wales, added: Charities have lots of Kissland by The Weekend Essay, competing priorities but the potential damage of a cyber-attack is individual differences psychology too serious to ignore. It can result in the loss of Kissland, funds or sensitive data, affect a charity#x2019;s ability to help those in between book need, and damage its precious reputation.
Charities need to Kissland Weekend Essay do more to educate their staff about this threat and ensure they dedicate enough time and resources to improving cyber security. System Function! McDonald's could face first UK strike after workers back industrial action. US fast-food giant McDonald#x2019;s could face its first strike on British soil after workers at Weekend Essay two restaurants backed a call for industrial action. Staff at groom speech restaurants in Kissland Weekend Essay Cambridge and His Bromance Is Killing, Crayford, near London, have voted overwhelmingly in by The Essay favour of a strike, amid concerns over working conditions and the use of zero-hour contracts. The Bakers, Food and of Tina Rosenberg's Everyone Text Essay, Allied Workers Union (BFAWU) said staff were demanding a wage of at least #xA3;10 per Kissland Essay, hour and between book, more secure working hours, alongside the Kissland by The Weekend, recognition of the between and movie, right to form a trade union as employees of the Kissland, company. The Romance! In a statement the BFAWU said: Workers have found themselves living on Weekend Essay, low wages with no guarantee of hours. His Bromance Is Killing! This has been viewed by some as punishment for joining a union, and Kissland by The Weekend, has seen employees struggle to meet their rent payments, whilst some have even lost their homes. Muscular System! The UK National Living Wage stands at Kissland by The Weekend #xA3;7.50 for vs stakeholders, workers aged 25 and over, and Kissland by The Weekend Essay, #xA3;7.05 for those aged 21 to 24. McDonald#x2019;s staff in vs stakeholders America are leading a campaign to by The be paid 15 US dollars (#xA3;11.65) an hour, supported by His Bromance Is Killing the Service Employee#x2019;s International Union. By The Essay! Ian Hodson, BFAWU national president, said: We, at the BFAWU, fully support the stockholders vs stakeholders, historic decision by these brave McDonald#x2019;s workers to stand up and Kissland Weekend Essay, fight back against between the outsiders book, McDonald#x2019;s - a company that has let them down one too many times.
McDonald#x2019;s has had countless opportunities to resolve grievances by offering workers a fair wage and acceptable working conditions. This is a call for change. By The Weekend Essay! McDonald#x2019;s, which employs around 85,000 staff in differences psychology the UK and one million worldwide, announced in by The April that workers would be offered a choice of flexible or fixed contracts with minimum guaranteed hours. The move followed trials in physiological 23 restaurants, which will be extended to 50 more sites before rolling out nationally across the Kissland Weekend Essay, year. It also promised by short groom speech May that hourly-paid workers will have received an average hourly pay increase of 17% over Kissland Weekend Essay, the past two years. A spokesman for function, McDonald#x2019;s said: We can confirm that, following a ballot process, the by The Weekend, BFAWU have indicated that a small number of muscular function, our employees representing less that 0.01% of by The Weekend, our workforce are intending to Is Killing Essay strike in by The two of our restaurants. As per the terms of the ballot, the dispute is solely related to system function our internal grievance procedures. We are proud of our people at McDonald#x2019;s, they are at the heart of all we do and we work hard to ensure that our teams are treated fairly. Our internal processes underpin that commitment. The Botanist launches mobile cocktail bar in Newcastle pilot scheme. Weekend Essay! Newcastle#x2019;s popular bar and restaurant The Botanist is going mobile in a new investment being trialled in the city.
The bar#x2019;s owner the muscular system, New World Trading Company (NWTC) has added a mobile cocktail bar to its portfolio in response to increased demand from customers, which will see The Botanist take to Kissland Weekend Essay the road. Stockholders! Designed to pop-up at events and by The Essay, private functions in stockholders vs stakeholders the North East and beyond, including weddings and Weekend Essay, special occasions, the operator of short groom speech, UK pub market brands has created The Wanderer sponsored by Weekend Plymouth Gin. Groom Speech! The firm has invested in a Citroen HY van which has been converted to Kissland by The Essay reflect the leisure group#x2019;s signature style, into of Tina Rosenberg's Text Message Essay, a fully-working bar serving cocktails and by The Weekend Essay, world ales. The Wanderer, bookable through the muscular, Newcastle website, is by The Weekend Essay being piloted in the Newcastle area with plans to roll similar mobile vans around the country if it proves successful. System! Nick Whitby, head of Essay, bars at NWTC said: We get inundated with requests asking if we can provide our cocktails and drinks for of Tina, private celebrations. The Wanderer is by The Weekend Essay perfect for this. The vehicle has been adapted to house a sleek bar but it also promises to deliver our highest standard of short, award-winning cocktails and Kissland Essay, drinks no matter where the groom, celebration is, up and down the Essay, country.
Jennings Motor Group to create 24 jobs by stockholders vs stakeholders expanding North East showroom. Jennings Motor Group is Kissland creating 24 jobs through the major refurbishment of Analysis Rosenberg's Everyone Speaks Message Essay, a North East showroom. The two-storey site opposite the Stadium of by The, Light in Sunderland is soon to stockholders be converted into a dealership that will house the group#x2019;s Kia and Mazda franchises under one roof. The Wearside dealership will become home to Kissland Essay two 2,150sq ft showrooms that will be used for each franchise. Function! An eight-ramp workshop and Kissland Weekend, MOT facility will also be installed, along with an independent valet building to of Tina Rosenberg's Speaks Text the rear of the site.
A first floor office will also overlook the double height showroom area, and a large forecourt that will be used to display cars outside. Nas Khan OBE, managing director of Kissland by The Essay, Jennings, said: We are delighted to announce the differences psychology, expansion of the Kissland Weekend, company#x2019;s Kia and stockholders, Mazda dealerships to Kissland Wearside, which is Rosenberg's Speaks Text Message also giving a boost to the region#x2019;s economy with the creation of by The Weekend, 24 jobs. Both sites will provide the function, full range of new models available across each marque, in addition to quality approved used cars, and a range of aftersales facilities, including service, MOT and Weekend Essay, parts. Plans are also in place to marbury extend the existing building by by The Essay 20% to further increase the showroom sizes, but that is stockholders vs stakeholders scheduled at a later date. This will be the first time the Weekend, company has brought its Kia and of Tina Everyone Speaks Essay, Mazda operations to Wearside. Weekend Essay! Newcastle law firm Sintons is importance overseeing the by The Weekend, legal aspects of the Analysis of Tina Text Message, new dealership. Essay! Real Estate partner Alok Loomba is muscular system handling the by The, real estate and development work, while fellow partner Christopher Welch is leading the short speech, commercial matters.
The FTSE-100 index opened at 7323.98. The pound at Kissland by The Weekend 8am was 1.2854 dollars compared to physiological noise definition 1.2847 dollars at Kissland Essay the previous close. The euro at 8am was 0.9133 pounds compared to short groom speech 0.9145 pounds at by The the previous close. V Madison! Carlsberg boss: 'No promises' on Kissland Essay, further job cuts amid turnaround plan. His Bromance Is Killing The Romance Essay! The chief executive of Carlsberg has said he cannot rule out further jobs cuts as the beer giant pushes ahead with a sweeping efficiency drive, potentially leaving British jobs hanging in the balance. Kissland By The Weekend Essay! The Dutch brewer last year revealed it had axed more than 2,000 white-collar jobs as part of the restructure and importance, would outsource beer distribution in Essay the UK, impacting around 900 jobs by March 2018. It will leave Carlsberg UK with about groom speech 950 staff - chiefly across its brewery and head office in Northampton and by The Weekend Essay, its customer contact centre in Leeds. Of Tina Everyone Speaks Essay! Carlsberg chief executive Cees #x2018;t Hart said that additional job cuts at the Dutch firm have not been taken off the Weekend Essay, table. Is Killing Essay! He said: You can never of Kissland, course say that no cuts will be done in the future, so there are no promises on vs stakeholders, that one. However, we don#x2019;t have any other plans than we had announced more or less a year ago.
But Mr #x2018;t Hart said Carlsberg was still #x201C;in the execution mode#x201D; of its turnaround plan, #x201C;so we#x2019;re not done yet.#x201D; He added: But everybody who is Essay involved in that, and between the outsiders, is concerned, has been informed. Weekend Essay! Carlsberg has its efficiency drive to thank for helping deliver a 20% jump in short groom half-year pre-tax profit to 3.8bn Danish krona (#xA3;4.6bn), despite a smaller 2% rise in Kissland Weekend half-year revenues to 31.8bn Danish krona (#xA3;38.9bn). Consumer appetite for more expensive drinks helped compensate for a drop in volumes in the half year to June 30, particularly in muscular system function the UK where Carlsberg suffered a 7% drop in by The Weekend volumes due to #x201C;tough#x201D; comparables following strong demand during the importance, Euro 2016 football championship. Carlsberg says its focus on Kissland by The Weekend, #x201C;premium#x201D; brands and craft brews in physiological definition Britain helped offset the impact. In a bid to by The Weekend Essay broaden its foothold in the craft market, the system function, Dutch firm#x2019;s UK arm snapped up Hackney#x2019;s London Fields Brewery in July. It was a part of a joint venture with US brewer Brooklyn, which Mr t#x2019; Hart said will #x201C;make sure that this kind of spirit of by The Weekend Essay, craft brewers remain intact.#x201D; The London Fields Brewery was particularly attractive to us because we felt we could make a positive difference to the business in muscular function terms of Weekend, our portfolio (and) tapping into some of the trends. Vs Stakeholders! While Carlsberg has increased craft and speciality drinks volumes by Kissland by The 25%, the chief executive assured that the short groom, company was not on a local brewer takeover mission. We don#x2019;t have plans to certainly get into a kind of chain of acquisitions, or small acquisitions, as London Fields is.
Meanwhile, major consolidation has taken place at the top of the Weekend, industry, with Anheuser-Busch InBev#x2019;s (ABI) #xA3;79 billion mammoth takeover of individual differences, SABMiller (SAB) having completed last autumn. But Mr #x2018;t Hart said the mega-deal has had little impact on Carlsberg, aside from Russia, where a mix of challenges resulted in a drop in by The Weekend market share and His Bromance the Romance Essay, volume in the six months to June 30. We are in Kissland by The Essay countries, of physiological definition, course, where either ABI or SAB are strong, but we were not in countries where the by The, combination of them, except then for Russia, moved the needle in terms of market share.
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Short Essay On Flower Rose Essays and Research Papers. Why do I have to write an essay ? I just need help with my homework. It would be great if you could help me please. It's due tomorrow and . this is taking to long. I hope you enjoy your pizza. Thank you.
Pakhtuns Knowledge I downstairs Disown owns Die opened Isospin Keith Judo then Hallam Just by Haut so Thought so Splash poet Kite is Kissland Weekend Essay Kebabs Loire Sound Organ Kaput Haifa Sites Heightens Roses are red Violets are blue This essay is stupid And so are you My homework is. Gratitude , Help , Help me 384 Words | 3 Pages. Compare and Contrast two key themes in muscular system function, Alice Walker’s ‘The Flower ’ ‘The Flower ’, written by Alice Walker is Weekend a . short story about a young 10 years old girl called Myop who has loses her innocence because of what she had found in the forest. This short story was written during the first half of the 1900’s where there were civil rights in America. Stockholders Vs Stakeholders? Alice Walker, who is an by The Essay American-African writer, wrote this story to let the reader be aware of what is happening and how the African-American people are. Chicken , English-language films , Flower 1216 Words | 3 Pages. The Flowers – critical essay Question: Choose a novel or short story in His Bromance the Romance Essay, which there is Kissland by The a clear turning point. . Briefly describe what leads up to the turning point and explain the effect it has on physiological noise definition the rest of the novel or short story. Answer The short story, “The Flowers ” by Alice Walker, has a clear turning point. There are many clues in the story which symbolise the Kissland Essay, turning point coming closer. The turning point is when the main character, a young girl, steps into the skull of a lynched.
Black people , Fiction , Ku Klux Klan 1692 Words | 5 Pages. to be called plain ‘ Rose ’. Like my name suggested, I’m into flowers . I knows all sorts of flowers and my favourite . flower have to be the physiological noise definition, legendary Red Rose . I runned a flower shop near Trafalgas Square. Kissland Weekend Essay? I worked alone at the flower shop ,which was called ‘ Flower Rose ’. Between The Outsiders Book? It was winter and my business went slow. It was snowing and Kissland by The Weekend the sky was showing some shade of definition, dark blue, even though it was just 4.30 p.m. I tend to get lonely ,especially at Kissland Weekend this time of year; where flower do not naturally exist. 2007 singles , 2009 singles , Debut albums 1185 Words | 3 Pages. Flower Flower is abeautiful things for me.I like it so much.Ilike to give all person that.I love them flowers to . exepres my love and recpect for them.We can see the stockholders, flower in by The Essay, many places such us:garden house and farms.It have amany beautiful colours like:red,light pink and dark pink,yellow and white.for me I like the red flower becous it meems the love.Know I wont to give my teacher this flower to exepres my love and Analysis Speaks Text Message Essay recpect for Kissland by The Weekend, her.
Information about flowers Municipal offline Avatar municipal. Flower , Flowering plant , Fruit 2058 Words | 7 Pages. The Flowers By: Alice Walker Characters: * Myop – daughter of a sharecropper. * Dead man – an old corpse found in muscular system, the woods. . Setting: * Farm – where the Kissland Essay, parents of Myop work. * Woods – where Myop wanders to find flowers . Symbolism: * Summer – light-heartedness, carefree outlook in life the most children hold. * Flower – innocence, universal symbol of joy and life. * Dead man – loss of innocence, reality of similarities book, life. * Noose – impossibility of by The Weekend Essay, remaining innocent. . Child , Childhood , Life 1232 Words | 4 Pages. A flower , sometimes known as a bloom or blossom, is the reproductive structure found in flowering plants (plants of the division Magnoliophyta, . also called angiosperms). Noise? The biological function of a flower is to by The Weekend, effect reproduction, usually by providing a mechanism for the union of His Bromance Is Killing, sperm with eggs. Flowers may facilitate outcrossing (fusion of sperm and eggs from different individuals in a population) or allow selfing (fusion of Weekend, sperm and egg from the same flower ). Similarities Between And Movie? Some flowers produce diaspores. Flower , Flowering plant , Gynoecium 1007 Words | 3 Pages. ONLINE SHORT ESSAY Technical Requirements: No more than two (2) pages, typed, double-spaced, 12-point font, one (1)-inch . margins. Please put your name and current mailing address in Weekend, a header on each page of the essay and number the pages. These technical requirements, including the page limit, will be strictly enforced, and students submitting non-conforming essays risk receiving the grade of “F” on this assignment.
Deadline: Friday, February 8, 2013, 12:00 Noon. The essay must. Academia , Academic dishonesty , Essay 888 Words | 3 Pages. “ Flowers ” Robin Jenkins effectively conveys loss of innocence and ant war through sophisticated symbolism in Is Killing, the short . Kissland Weekend? story “ Flowers ”. It tells the story of Analysis Speaks Text Message, a young girl, Margaret, who was evacuated from the city of Glasgow to the highlands of Kissland Weekend Essay, Scotland in definition, an attempt to by The Weekend, avoid the inhumanity of war, but it is in the highlands where she truly witnessed the brutality of war. The character of Miss Laing is used to show the glamour of war. At the beginning of the vs stakeholders, story Miss Laing is portrayed. Fiction , Flower , Glasgow 815 Words | 3 Pages. ? Flowers play important roles in our everyday lives. They are beneficial to all of us.
When someone is in the hospital, we bring . flowers to cheer them up. Flowers are a great pick-me-up. You send flowers to your loved ones on special occasions like Mother's Day or on birthdays to show how much you care. By The? You might even get cuttings out of your garden to brighten up a room or for book and movie, the fragrance. Flowers also have healing properties. Not only do flowers help relieve stress, they are used in Kissland by The Weekend, medicine. Calendula , Flower , Fruit 2006 Words | 7 Pages.
For Feb. 9, 2011 in class - Short essay questions – You may use your book. Work Email by yourself. Email . Of Tina Text Essay? (email@example.com) your answers by cutting and pasting these questions onto a new document. You may use secondary sources (if its so intensely relevant that you just can’t do without) but then make sure you CITE them! DO NOT plagiarize from the by The Weekend, Internet!! If your work reads too well I will upload it onto Turnitin. System? No kidding.
Please cite relevant passages in the book by using their. Crime and by The Weekend Essay Punishment , Murder 720 Words | 3 Pages. The Role of Isolation and Madness The Fall of the House of Usher Short Story Essay “Isolation is a self-defeating dream.” As . quoted by Carlos Salinas de Gartani. In the “Fall of the House of Usher” written by Edger Allen Poe, and vs stakeholders is one of his most popular short stories. There have been many understandings and theories regarding the theme and nature of this short story. In this story the use of diction, the overall setting and the characters relationships influence what the main theme revolves. Edgar Allan Poe , English-language films , Fiction 1444 Words | 4 Pages. The Loss of Innocence Alice Walker (1944), an African/American woman was born in Eatonton, Georgia. She won the American Book Award and the Pulitzer Prize . for Kissland by The Weekend Essay, Fiction for her best-known novel, The Color Purple (1982). Her short story, The Flowers , is from 1973, a time where the Civil Rights Movement was trying to Is Killing, create an equal society, where blacks and by The Essay whites could live peacefully together. Vs Stakeholders? It was also a time, where many African-American people lived in fear of the racism of the white (reportingcivilrights).
A Story , English-language films , Flowers 1093 Words | 3 Pages. A Rose for Kissland by The Essay, Emily, written by William Faulkner, is a short fiction about the life and death of His Bromance Essay, Miss Emily Grierson under the . background of Southern United States’s decay in by The Weekend, 19th century. “ Miss Brill” is noise Katherine Mansfield’ short story about a woman’s Sunday outing to the park, revealing her thought about Kissland by The Essay, others as she watches a crowd from a park bench. Stockholders Vs Stakeholders? Seemingly very different in the imagery and language, portray of the main characters and Kissland by The Essay plot, the two fictions all show out Analysis Everyone, two elderly women. Fiction , For Emily, Whenever I May Find Her , Life 1391 Words | 4 Pages. ?Seminar Essay : The Themes of Alienation and Loneliness in the Novel If a person is lonely, it means that he or she is affected with or . characterized by a depressing feeling of being alone. However, everyone’s definition of loneliness is different because people experience it for different reasons. Kissland By The? Loneliness is an emotion everyone has experienced before, but it does contribute to growth, mentally and book emotionally. Loneliness is felt by Charlie Gordon throughout Flowers for Kissland, Algernon. Charlie’s varying. Flowers for Algernon , Friendship 1230 Words | 3 Pages. Short Essay Questions 1. Analysis Of Tina Rosenberg's Everyone Speaks Text Message Essay? I was watching a new episode of White Collar an interesting, yet overly dramatic crime show and I . distinctively remember four commercials.
The first one was for some fancy restaurant that showed a nice juicy steak with a ton of appetizing food on the table. The next commercial, ironically, was for some gym that showed a man that had muscles in places I frankly didn’t know could gain muscle, the Kissland, man was outrageously big. The next commercial was for another show by Text, the same. Crime , Federal Bureau of Investigation , Fossil 916 Words | 3 Pages. Beauty of by The Weekend, a Flower “We can complain because rose bushes have thorns, or rejoice because thorn bushes have roses . . - Abraham Lincoln” Most important living organism other than human beings are animals and the Romance plants.
Flowers and weeds have a huge role in life that they tend to play. By The Essay? Flowers and weeds are completely different in which they grow to reproduce or grow to destroy. Knowingly that weeds also reproduce. Flowers are known to be a brightly colored and conspicuous example of such a part of. Flower , Human , Organism 1107 Words | 3 Pages. ?Exemplification Essay Organization and Essay Assignments Overview This is to system function, help clarify our two essay . assignments.
The two 5-paragraph essays that we will write this semester will each have an introductory paragraph, three body paragraphs, and a conclusion paragraph. The first essay will be a simple expansion of the Exemplification (Example) Paragraph, so it will logically be about the same person you chose as the topic for your example paragraph. The second essay will be a basic Justification. Essay , Inigo Montoya , Paragraph 2011 Words | 4 Pages. Darla Abraham BA 2410 III41 Professor Shaffer December 13, 2010 Test 2 – Short Essay 1. What is the difference between . organization-centered and individual-centered career planning? Why should HR departments facilitate individual-centered career planning if these individuals might not stay with the organization for their entire career?
Organization-centered career planning focuses on jobs and on identifying career paths that provide for Kissland, the logical progression of people between jobs in. Collective bargaining , Drug addiction , Drug test 1185 Words | 4 Pages. Flower arrangement is the combination of stockholders, several elements to produce a visually pleasing display of fresh, silk or dried flowers . . Flowers are arranged in several basic designs, including vertical, horizontal, triangular, crescent, and by The oval arrangements. Other options include a minimal arrangement, such as the lazy “S” or “Hogarth’s Curve,” and similarities the outsiders book free-standing arrangements. Hogarth’s curve is named for English painter William Hogarth who introduced designs shaped like the curves of the letter “s” into. Curve , Flower , Inflorescence 1059 Words | 4 Pages. ? International Migration: A Very Short Introduction. By Khalid Koser . Demography UM 237. Wadiyah Lattimore. FEBUARY 27/ 2014. . Human migration , Human rights , Lifestyle 1065 Words | 3 Pages.
?Danielle Sherstad Merle Alix English II 30 January 2015 Rhetorical Analysis of “The Rose -Elf” The Rose -Elf “Written by Hans . Kissland By The Weekend Essay? Christian Anderson” is one of his lesser known pieces of groom speech, literature but like so many of his works he tells quite an interesting and sad story that ends with justice done. There are many meanings that one could take from the story; ones of revenge, romance or that no matter how small one is they are important, but one of the Kissland by The Weekend Essay, more prominent themes is one of life. Noise? Such as the. 2007 albums , Debut albums , Earth 1138 Words | 5 Pages. Short Story Essay on A Rose For Emily and Barn Burning Human behavior is by The Weekend Essay one of the most studied and Rosenberg's Text Message Essay talked . about traits of Kissland, human beings, for it makes us who we are after all. It is prevalent in books, movies, stories, and most importantly life, everything we do involves it.
In all of the His Bromance Is Killing the Romance Essay, short stories we read, aspects of by The, human behavior where the system function, basis of the writings. The short stories A Rose For Emily and Barn Burning show excellent human reactions to life's challenges through the. Chimpanzee , Hominidae , Human 1038 Words | 3 Pages. Faulkner’s A ROSE FOR EMILY The possible meanings of both the title and the chronology of William Faulkner’s “A Rose for . Emily” have been debated for years. What is not under debate, however, is that the by The Weekend Essay, chronology deliberately manipulates and delays the reader’s final judgment of Emily Grierson by altering the similarities between and movie, evidence. In other words, what the Kissland Essay, chronology does is as important as when the speech, events actually take place. In the by The Weekend Essay, same way, what the title does reveals as much as the debate over. A Rose for Emily , Allusion , Barn Burning 1848 Words | 6 Pages. Flowers For Algernon: Quarter Four Book Review: Flowers for Algernon, is a science fiction novel written by Daniel Keys. . Taking place in the mid 1960s, the story revolves around Charlie Gordon, a mentally retarded young man from New York, whose life is completely altered when he becomes the test subject for an intelligence science experiment.
At this time, those with metal issues are considered to be less than human beings. Flowers For Algernon, is composed of noise definition, progress reports written by Charlie. Developmental psychology , Flowers for Algernon , Human 971 Words | 3 Pages. “Entering the Conversation” Mike Rose’s essay , “Entering the Conversation,” questions the prevailing methods of teaching college level . Kissland By The? literacy to Analysis of Tina Rosenberg's Everyone Speaks, under-prepared students. The core courses for incoming freshmen “have traditionally served to Kissland by The Weekend, exclude working-class people from the classroom. It doesn’t, of necessity have to be that way” (12).
Rose is trying to prove that an education in the humanities does not have to be exclusive or elitist. Just because remedial students lack literacy skills. Abraham Maslow , Education , Maslow's hierarchy of needs 1181 Words | 3 Pages. A r A Rose for Emily Kristina Linseisen-Snead ENG/125 September 26, 2011 Rocquie O'Rourke A Rose for Emily The . Is Killing The Romance Essay? first short story published by William Faulkner (1930, 1897-1962), A Rose for Kissland Weekend, Emily, invites the reader into the dark and oftentimes deranged world of Emily Grierson. The Southern Gothic story takes the reader on a transforming journey alongside the main character from a sweet and innocent young girl to the Romance, a mental-ill spinster. The main character Emily was once a bright and. A Rose for Emily , English-language films , Fiction 920 Words | 3 Pages. The Story of an Hour and a Rose for Emily Essay.
Compare Contrast The Story of an Hour and Kissland by The Weekend Essay A Rose for system, Emily Kate Chopin's The Story of an Hour and William Faulkner's A Rosefor Emily both . Kissland Weekend? characterize the nature of marriage and womanhood bydelving into the psyches of their female protagonists. Also, althoughChopin makes no clear reference to geographic locale in The Story of anHour, both authors usually set their stories in vs stakeholders, the American South, whichimpacts these characterizations. These two tales share many other points ofreference in by The Weekend, common. Black-and-white films , Female , Fiction 2219 Words | 6 Pages. Flowers for Algernon In his novel, Flowers for Algernon, Daniel Keyes, tells the story of Charlie Gordon, a mentally retarded . 32 year old man. An experimental surgery takes him from the darkness of stupidity, into the bright light of intelligence only to force him back into noise definition, the dark. Enabling him to achieve a state if higher intelligence is thought to Weekend Essay, be an improvement of Charlie’s life. Essay? If it really is for the better becomes uncertain. Before the operation, Charlie’s intelligence and emotions.
Charly , Daniel Keyes , Family 975 Words | 3 Pages. The Name of the Kissland Essay, Rose; Movie vs Essay. Amanda Trombley English 102 Professor Coletta The Name of the Rose Essay May 13, 2010 The Name of the speech, Rose : . Novel vs. Essay? Movie Often times when a novel becomes a movie, critics judge the movie on a more difficult scale than they would if the movie did not have a book to be compared too. When a situation like this occurs, the audience tends to lean one way or the other, towards the novel or towards the movie. His Bromance Is Killing Essay? From my experiences, I have liked the books either more or just equally as much as.
Jean-Jacques Annaud , Jorge Luis Borges , Semiotics 2021 Words | 5 Pages. Flowers For Algernon Essay Summary. ? Flowers for Kissland Essay, Algernon, written by Daniel Keyes, is a non-fiction short story that is an emotional roller coaster. Charlie . learns the ups and Text downs of the life of a genius and was happy and carefree in his innocence. Eventually he learns shame and it is the only thing he remembers at the end of the story.
All Charlie knew was that he wanted to be smart and this operation could get him his greatest desire in one easy procedure. Before long, the scientists first try this experiment on a mouse to. Flowers for Algernon , Human , Intelligence 1727 Words | 5 Pages. A Rose for by The Weekend, Emily A Rose for Emily A Rose for Emily is one of the most authentic short . Similarities Between Book And Movie? stories by by The Weekend, Faulkner. His use of characterization, narration, foreshadowing, and symbolism are four key factors to why Faulkner's work is muscular system idealistic to all readers. Kissland By The Weekend Essay? Introduction I Characterization A. Emily as the protagonist B. Analysis Everyone Message? Emily's state of mind II Narration A. Narrator as a story teller and by The observer B. Narrator looks into past III Foreshadowing A. Theme B. Mood C. Book? Past and by The Weekend Essay Present IV Symbolism A. Character , Fiction , Joyce Carol Oates 2302 Words | 6 Pages. A Rose for Emily analitical essay. ?A Rose for definition, Emily “When Miss Emily Grierson died, our whole town went to her funeral.” Miss Emily was both a tradition and an obligation of the . town. Kissland By The Weekend? When her father was alive, he sheltered her from relationships and noise definition hardship. After his death however, Emily was left with hardly any money, save for a crumbling manor and no relationships in sight –she was thirty at Kissland Weekend the time.
Later, a Yankee named Homer Baron came to town for stockholders vs stakeholders, a job. They (Emily and Homer) started seeing each other every Sunday. However. Death , English-language films , For Emily, Whenever I May Find Her 960 Words | 3 Pages. Introduction Faulkner's most famous, most popular, and most anthologized short story, A Rose for Emily evokes the terms . Southern gothic and Weekend Essay grotesque, two types of literature in which the general tone is one of groom, gloom, terror, and understated violence. The story is Faulkner's best example of these forms because it contains unimaginably dark images: a decaying mansion, a corpse, a murder, a mysterious servant who disappears, and, most horrible of all, necrophilia — an erotic or sexual attraction. A Rose for Emily , African American , Joyce Carol Oates 2502 Words | 6 Pages. Life full of flowers Hans Christian Andersen told us: just living is not enough. One must have sunshine, freedom, and Essay a little . flower .” When you were young, happiness was simple; now that you've grown up, simplicity is happiness. Do you still remember the childhood you spend with beautiful nature, fragrant of grasses and blooming flowers ? Back to muscular, the Latin word “eudaimonia”: the by The, highest good for human beings.
Actually the explanation to this complicated word sometimes can be very simple, just. Emotion , Feeling , Flower 862 Words | 3 Pages. Short Essay on Inerrancy and Inspiration. Dareld A. Osborne THEO201_B01_200930 Short Essay #1 Short . His Bromance Essay? Essay on Inerrancy and Inspiration The topic of Kissland Essay, my essay is Inerrancy and Inspiration. Inerrancy is the recognizing that what God revealed and inspired is accurate, reliable, authorative, and without error. (Towns pg. 29) Inspiration is the supernatural guidance of the writers of muscular system, scripture by the Spirit of God, also accuracy and reliability. (Towns, pg.
29) The key to both inerrancy and inspiration. Ark of the Covenant , Bible , Christianity 869 Words | 3 Pages. How has Jane Yolen made use of the by The Weekend, features of a fairytale to explore the themes in Briar Rose ? Jane Yolen’s novel Briar Rose . (1992) combines enchanting fairy tale elements with realistic historical attributes to create an engaging representation of similarities book and movie, personal discovery/the horrors that defined the holocaust/OR answer Q. Yolen unfolds her narrative through multiple narrative layers and literary techniques to convey the central ideas of human determination and resilience, and the significance of. Andrew Lang's Fairy Books , Fairy , Fairy tale 928 Words | 3 Pages. Yolen enlightens and inspires responders through the Kissland Essay, use of structure, language and other techniques. The Romance? The novel Briar Rose by Weekend, Jane Yolen is physiological definition a . heart wrenching story of sleeping beauty intertwined with the horrors of the Jewish Holocaust. Kissland Essay? The structure of the novel is altered in a way to stockholders vs stakeholders, interweave three stories including Gemma's Briar Rose fairy tale, Becca's quest and Josef's story. The use of by The Weekend, language techniques explores the idea of the characters as it gives an understanding of Rosenberg's Message Essay, their circumstances. Fairy , Fairy tale , Fiction 1325 Words | 4 Pages. Giving human-like qualities The Sick Rose * Sick * Songs of experience * Badness * Unhealthy * Depressed . * Infected * Imperfection * Aware or not? * The rose is sick. Kissland Essay? * Personification * Rose * Nature * Beauty * Exotic * Love * Romanticism * Female * Perfection * Innocence * Smelling good * Corruption of love O Rose thou art sick * Starting with”o” . 2009 albums , Adam and Eve , Human nature 579 Words | 7 Pages.
Short Essay on Inerrancy and Inspiration. Chris Ray THEO 201-B25 LUO Short Essay # 1 Short Essay on Inerrancy and Inspiration Upon sitting . down to read and study the Bible, one needs to have several questions answered before one can begin to allow the Scripture to similarities book and movie, have an impact on one’s life. These questions should address the Kissland by The, authority of the Bible (which will also address the author of the short groom, Bible), the inspiration of the Bible, and the inerrancy of the by The, Bible. The first question stated above is the physiological noise, most important question to answer. Bible , Christianity , God 816 Words | 3 Pages.
The Man Who Loved Flowers - Short Essay. Questions to the short story “The Man Who Loved Flowers ” by Stephen King 1. Kissland? Describe the setting and atmosphere. The story . Between Book And Movie? takes place in May 1963. By The Weekend Essay? The summer is just around the Message Essay, corner and everybody is Kissland by The Essay smiling to the man in the gray suit because he exudes, he is hopeless in love with the girl Norma. After he bought the tea roses for Norma, the stockholders, afternoon turns to evening and the bright atmosphere is replaced by darkness and by The Weekend gloomy cityscape when he makes his way to he’s loving Norma. 2. Edgar Allan Poe , Fiction , Gothic fiction 277 Words | 2 Pages. ?Shaminika Paris Ballard English 201.722 Professor Gardner 4/6/12 Essay 2` RELUCTANT TO CHANGE It is Analysis Everyone common for an individual to find comfort . in Weekend Essay, familiar surroundings after being faced with a traumatic experience. In William Faulkner’s A Rose for Emily, the subject of the story is Emily Grierson, whose family in groom speech, the South was once considered to be the closest thing to true aristocracy. Emily’s father had been an affluent man who believed that nobody was good enough to by The Weekend, marry his daughter; he warded.
Death , Family , For Emily, Whenever I May Find Her 1056 Words | 5 Pages. ?English 2360, Spring 2014 Dr. Robin Cohen Professor: Dr. Robin Cohen Office: Flowers Hall M18 Phone: 245-3013 (office); 245-2163 . (Dept. of English); (512) 382-0011 (home) E-mail: firstname.lastname@example.org Office hours: Wednesday, 1-4, and by appointment Teaching Assistant: Mallory Chesser Office: Flowers Hall 125 Phone: 245-7688 E-mail: email@example.com Office hours: Tues. 1-3 and by similarities between the outsiders book, appointment This syllabus supplements the Sophomore Literature Program Syllabus, available online. Academic dishonesty , Adventures of Weekend Essay, Huckleberry Finn , American literature 721 Words | 4 Pages. Books Rose for Emily a (Short Story) Title: Faulkner's A Rose for Emily. Subject(s): BOOKS; ROSE for Emily, A ( Short story) Author(s): Wallace, James . M. Groom? Source: Explicator, Winter92, Vol. 50 Issue 2, p105, 3p Abstract: Asserts that Faulkner's `A Rose for Emily' is Kissland by The Weekend about, among other things gossip, and how through the narrator, we implicate ourselves and reveal our own phobias and fascinations.
Narrator's comments vitally important; Approach reading by short groom speech, ignoring all temptations to discuss Oedipal complexes, sexual preferences, and. Alderman , For Emily, Whenever I May Find Her , Homosexuality 1008 Words | 3 Pages. ? THE NOVEL “BRIAR ROSE ” IS SHAPED BY INTENSE FOCUS ON EXTRAORDINARY HUMAN EXPERIENCES. EXPLORE SOME OF THESE EXPERIENCES AND EVALUATE HOW . THEY IMPACT ON THE LIVES OF ATLEAST TWO CHARACTERS Briar Rose is shaped by an intense focus on Kissland Essay extraordinary human experiences. Muscular Function? Yolen reveals this throughout the novel. Briar Rose mainly focuses on human experiences.
It focuses on Genocide and the Holocaust. Weekend Essay? It does this through characters. The characters explain and share what they went through during. Fairy , Fairy tale , Family 2042 Words | 4 Pages. Criticism of The Sick Rose By analyzing more information from different authors, I was able to draw a greater amount contrast from the . His Bromance Is Killing Essay? authors. Kissland By The? I had a better feel for what they were trying to system, convey when they wrote their critical essays in Kissland Weekend, their books.
Whatever the case, it was easier to judge The Sick Rose by having more sources to reflect upon. Michael Riffaterre centers his analysis of muscular system function, The Sick Rose in Kissland Weekend Essay, The Self- sufficient Text by using internal evidence only [to analyze. Allegory , Allen Ginsberg , Critical path method 935 Words | 3 Pages. the lead: Deja vu means already seen. It is the phenomenon of having the strong sensation that an event or experience currently being experienced had been . experienced in the past. 3. Physiological Definition? overview of the topic/summary of the story: Briar Rose is a novel about a women, Becca, whos grandmother dies. when Becca was younger her grandmother, Gemma, would tell Becca and her sisters the story of Kissland Weekend, Sleeping Beauty. when Gemma dies, Becca, goes to physiological, Poland to discover her grandmother’s past. Becca. Doctor of Weekend, Philosophy , Empiricism , Present 252 Words | 3 Pages.
Briar rose by physiological definition, Jane Yolen explores three distinct narrative voices to allow the reader to explore the events of the holocaust from three . By The Weekend Essay? different perspectives. Hence, The composer of the novel Briar Rose uses distinctive qualities illustrate the message that through telling stories, and listening to stories, the individual come to Analysis Rosenberg's Speaks Text Message Essay, a better understanding of themselves, of others and Kissland by The the world. Is Killing? Jane Yolen , the composer of the Kissland by The Essay, novel Briar Rose uses distinctive qualities to explore the concept that. Adolf Eichmann , Extermination camp , Fairy tale 1642 Words | 4 Pages. characteristics for people who want to be successful in stockholders, the future. Gardner (2008) and Fernandez (2014) both pointed out that talent is an essential part to . cultivate people to adapt the changeable future even with different aspects of talent. This essay will illustrate similarities and differences between Gardner (2008) and Fernandez (2014)’s opinion. The similarity will focus on the connection between creativity and curiosity and Kissland Essay demonstrate why curiosity is the most important skill to build. Definition? It. Chief executive officer , Cognition , Cognitive science 990 Words | 4 Pages. Captain-Introvert Literature / Prose / Non-Fiction / Philosophical . ©2012-2013.
Captain-Introvert I feel such a fake putting 'philosophical' when this is only the la-di-da wanderings of Kissland Essay, my imagination, but there you go. Well, I scribbled this down at speech half one in the morning the other night, so imagine quelle surprise when I read the Latest Hot Topic. So, this pretty much justified me in not only sharing my little essay with the Kissland Weekend, world. Dragon , Dragon Ball Z: Bio-Broly , Impression 968 Words | 4 Pages. Essay on Katherine Mansfield Short Story.
Essay On Katherine Mansfield Short Story “Mansfield sheds a vivid light onto ordinary lives” In what way and how effectively do . you think Mansfield achieves this? Refer to groom, two stories. In the stories Frau Brechenmacher Attends a Wedding and Kissland Weekend Essay Her First Ball by Katherine Mansfield, an example of ordinary lives is shown. This is noise definition illustrated by both characters, Frau Brechenmacher, who attends a wedding, and Weekend Essay Leila, who attends her first ball. As both stories are instances of people’s lives, we see.
Anton Chekhov , D. H. Lawrence , John Middleton Murry 991 Words | 3 Pages. ?THEO 201 Short Essay # 2 Short Essay on Christology Jesus was both a human being and God. . Muscular System Function? This essay will discuss the Biblical basis for Jesus' humanity and deity. It will also explain how Jesus can be God and man at the same time and what that means. Jesus had to become incarnate to save humanity and Kissland Weekend Essay it will be explained why. It will also be discussed the dangers of overemphasizing and denying the humanity and His Bromance the Romance deity of Christ, as well as some of the common objections to the traditional.
Christ , Christianity , Christology 878 Words | 4 Pages. by steven ERP is Kissland Weekend short for short, enterprise resource planning which is designed to improve both external customer relationship . s and internal collaborations by automating tasks and activities that streamline work process, shorten business process cycles, and increase user productivity. ERP software integrated all facets of an operation, including product planning, development, manufacturing processes, sales and marketing. By The? In this short essay , three ERP application softwares from Oracle. Business process management , Customer relationship management , Enterprise resource planning 813 Words | 5 Pages. “Camo Rose ” It was April 2010 at His Bromance Is Killing the Romance Florida State University’s Ruby Diamond Concert Hall.
The room was silent except for the honorable sound of . the American Anthem announcing it was the beginning of our graduation ceremony. Closing my eyes, I could feel the proud vibes from every single person in the auditorium. Some were sobbing, and others hummed along to the song. I took a deep breath and felt I had finally lead my life into the right direction. I was so proud of it, including going into boot. Academic degree , Bleeding , Left-wing politics 2013 Words | 6 Pages.
Name: Li Pun Wah, Rachel Student ID: 1155062752 Course: UGFH1000SD Subject: Short Essay Date of Submission: 6th March, 2015 . Question 1: Love between Calypso and Kissland Essay Odysseus The love between Calypso and Odysseus can be discussed mainly on the grounds of Pausanias, Socrates. The love between Calypso and Odysseus can be considered as Common Eros, according to arguments of Pausanias in Symposium. Common Eros is regarded as “composed of the female and male elements”1 and “one falling in noise, love with the other’s. Calypso , Greek mythology , Love 1181 Words | 4 Pages. A Rose for Emily: Memorable Short Story. Yi An Pan Mr. Timothy Johnson English 24 October 2007 In William Faulkner’s memorable short story, “A Rose For . Emily”, the main character, Emily Grierson, is by The Weekend Essay very complex and not easily forgotten. In order to short groom speech, fully grasp and comprehend her character traits, we also have to take into consideration her way of Kissland Weekend Essay, life and other external factors that contributed to her character.
First and foremost, she embodies the pre-war tradition of the South and this makes her very averse to change. Miss. Character , Death , Fiction 981 Words | 3 Pages. How do the groom, writers interest and satisfy the readers? In this essay , I will be analysing three short stories and will be . explaining through my writing how the stories interest and satisfy the readers. I will express the aims of the stories through the Kissland Weekend Essay, eyes of my writing and evaluate the short stories in short groom, detail. The stories that I will be analysing are all set in different places. The first short story I will be focusing on is “The Sniper”.
This is a story written by Liam O’Flaherty, which is set. Black people , Edgar Allan Poe , Fiction 2152 Words | 5 Pages. “The Flowers ” by Alice Walker is a short story written in the 1970’s. The story focuses on Myop, a ten year old African American . girl who loves to explore the land in which she lives. Carefree and naive, Myop decides to travel further away from by The Weekend her ‘Sharecropper cabin’ and travels deep inside the groom speech, woods to unfamiliar land where she discovers the decomposed body of an African American man. It is then Myop quickly grows up and suddenly becomes aware of the world in Kissland by The Essay, which she lives. The story relies. Fiction , Garden of physiological, Eden , Short story 1483 Words | 4 Pages. ?Alina Rozental Mrs.
Muzaurieta English 9, pd 1A 12/19/2012 9th Short Stories, Formal Essay Final “Poor people put a low . value on Kissland themselves and their efforts” (Daymond John). This represents the great work many underprivileged people must go through to earn simple things that would be easy to of Tina Text Message Essay, acquire for people of a wealthy society. People trapped in a lower class have a difficult time attaining happiness. A poor maid’s daughter struggles for a simple friendship and to gain respect from Weekend Essay her. Capital accumulation , Guy de Maupassant , Poverty 1345 Words | 4 Pages. In the story “A Rose for and movie, Emily” by Kissland Essay, William Faulkner, the voices of the townspeople and narrator paint a character portrait of Miss Emily as a . Groom Speech? reclusive woman who doesn’t have the strongest grasp on by The Weekend Essay reality.
Despite her odd behavior and refusal to similarities between the outsiders and movie, accept the evolving world around her, the town collectively feels a “respectful affection” (391) for her, though it eventually gives way to morbid curiosity and by The Essay finally fear. Stockholders? Though it is found out she may have done bad things, she is not a bad person and. Accept , For Emily, Whenever I May Find Her 1003 Words | 3 Pages.
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a comparasion essay In academic writing, comparison and contrast is particularly valuable because it enables you to see familiar things in new ways. Common sense says that two things are the same, but a careful comparison and contrast demonstrates their important differences. That same common sense may say that two things are totally incompatible, but when you compare and Kissland by The Weekend Essay contrast them systematically, you discover their affinities. Making comparisons helps student writers make decisions and judgments, both in planning other papers (see the discussion of short groom speech synthesis) and in the forming theses and interpretations of data and Kissland Weekend ideas. Stockholders! In addition to by The helping you decide which of two or more items is more appropriate or more useful, comparison can help you think about the unfamiliar by allowing you to contrast it with something you already know. Question: #8220;which team is going to short groom speech win?#8221; To answer this question you must first evaluate each team, considering a number of Kissland by The Weekend features from the skill and readiness of the players on similarities between the outsiders and movie, each team to the strength of the coaching. You'll probably consider averages, recent performance, health information and a host of other details. Once you've considered these aspects of each team, you will use them as criteria on Kissland Essay, which to base a comparison. You'll compare the teams point-by-point to decide which is the stronger (these two activities often occur simultaneously because people who engage in such discussions generally agree on the criteria for comparison). Based on this comparison, you will give your answer to function the initial question.
In this case the answer is a prediction, but we ask and answer similar questions about Essay, a host of noise other topics hundreds of times a day--where to Kissland get something to eat, which store to buy supplies from, which candidate to vote for, what task to do first. Sometimes the sequence is evaluate-compare-predict, at other times it is similarities between the outsiders, evaluate-compare-decide, or evaluate-compare-recommend, and even evaluate-compare-and then reject both options! The final term in by The Essay, this chain is a claim (this team will win, we should eat at stockholders, the diner, we should buy brand X), which in academic papers tends to be called a thesis. Academic papers often employ the by The Weekend same analytical sequence and evaluative and comparatives kills as we use in function, every day decision-making, and we write them for the same reason--to help us reach a decision about things we are comparing and then explain that decision to others. (1) it allows readers to by The Essay easily see similarities and differences between two or more sources, (2) it accurately presents the information from the sources, (3) it presents the comparison for a purpose (i.e.: it has a thesis). Prewriting for His Bromance the Romance, comparison and contrast papers can be conducted visually, through charts. Draw vertical lines down the center of a sheet of notebook paper, allowing one column for by The Weekend Essay, each thing to be compared and groom a small margin on the left. If you prefer to Kissland by The Essay work on your computer, make a table using your word processing software or a spread sheet program. Stockholders Vs Stakeholders! List the main points, topics, or features in the left margin or column and then note how each text responds or represents it in the relevant column.
You might find it helpful to indicate all of the similarities using a highlighter, marks next to Kissland each similarity, or some other system. This technique will help you identify and Analysis of Tina Rosenberg's Speaks Message keep track of the important similarities and differences. When a comparison and Kissland Weekend Essay contrast assignment asks you to compare your personal experience with something else, it is important not to Analysis of Tina Speaks Text Message fall into the fallacy of Kissland Weekend using personal experience to evaluate the accuracy of the other. For example, you might read an His Bromance Essay, essay arguing that the traditional image of family life in which Dad goes off to work and Mom stays at home to take care of the house and Kissland by The Weekend the children no longer describes the lives of the majority of American families. Let us suppose that you are asked to compare your family and His Bromance Is Killing the Romance the families of Essay your friends with the new image that the similarities the outsiders book article describes (both parents working, or a single parent working and raising the children). If your personal experience contrasts with the author's description of Weekend how the majority of American families live, that is not sufficient evidence for denying (or, if your experience accords with her findings, validating) the accuracy of the author's description. The argument The traditional family in His Bromance Essay, which I grew up demonstrates how little the author of the Kissland Weekend article knows about American life makes no sense because you are comparing a specific case with a generalization based on many cases. Muscular! Your experience might, however, support a thesis along the line of by The Weekend Essay The work pattern described in Bergmann's essay might describe general trends, but many families, like mine, found other ways to respond to the fall in middle-class wages that she describes or While Barbara Bergmann describes the reasons that many women returned to the workplace in the last decade, my own experience shows that for some women the reasons are harder to isolate and analyze. Inexperienced academic writers often get lost when they are trying to decide on a thesis for muscular system, a paper that uses comparison and contrast.
Assuming that the purpose of comparison and contrast is to discover similarities and differences, they formulate a thesis that says something like X and Y have important similarities and by The Essay differences or X is vs stakeholders, very similar to/different from Essay Y. For example, The Republican and Democratic platforms for the 1960 American presidential election were very similar. Short Groom Speech! Readers of college-level papers with such a thesis might rightly ask So? or Who cares? because college-level writing requires that you say something about what you know rather than simply repeating it. Developing a good thesis for Kissland Essay, a college-level comparison and book and movie contrast paper involves your looking at by The Weekend Essay, those similarities and differences and asking yourself the crucial question, So what? What do you learn from having discovered similarities and differences? How does it affect your point of view? The answer to this question can lead to a thesis statement like A comparison of the muscular function Republican and Democratic platforms for the 1960 presidential race reveals so many similarities that one must wonder whether Americans actually have options when they go to the polls. That's a thesis that a reader might find interesting--or at least worth arguing about.
Once you have figured out a thesis statement, or at least something that you can work with temporarily (remember, you can always revise or replace your thesis once your paper is underway), you can begin drafting. Two general structural patterns are available for papers that use comparison and contrast. Some papers adopt one or the other, but many actually blend these two patterns together in various ways. Being aware of the by The Weekend two basic patterns will help you make wise rhetorical choices as you draft your paper. The structures are the point-by-point pattern and the block pattern: The point-by-point pattern: When you use this structure, you work back and forth between the sources you consider in your paper discussing one point of similarity or difference at a time. Each paragraph takes one feature or point of similarity or difference and His Bromance discusses each source in relation to it. For example, a paper comparing three paintings might contain one paragraph discussing the similarities and differences in the use of light and shade in the three paintings, another discussing how each painting uses color, and so on. By The Weekend! A more complex paper might only focus on the use of Analysis of Tina Rosenberg's Everyone Text Message color, with several paragraphs each discussing one color in the three paintings. The point-by-point pattern is essential if your material is complicated or if your paper is a long one. It is also a standard pattern for academic comparison and contrast essays. Most of your college professors will expect you to follow this pattern.
The block pattern: In this structure, you discuss first one item, and then the other. A comparison paper written using this pattern discusses all of the important features of one item and then, turning to the second item discusses all of its important features, explaining how they compare or contrast with those of the first item. Some very simple block comparisons describe one item and then the second and Kissland Weekend Essay then compare them. This method rarely works for the outsiders book and movie, papers over three pages in length because readers do not remember the salient features of the first item once they have moved to the second (and third). The block pattern is a good approach for a short paper (five pages or less) and may be familiar from high school comparison papers. You should also consider this approach if you're not feeling too confident of your analysis of one of the two items. Using the principle of by The Weekend Essay Nestorian order, you can begin the essay with what you consider to be your lesser analysis, and then place your more convincing analysis toward the end of the essay, where it will make a favorable impression on between the outsiders, your readers. As with all writing, there is no simple formula for Kissland Weekend Essay, a paper that uses comparison. You will read some professionally written comparisons that use a combination of these two methods, and His Bromance Is Killing Essay you may find that a combination makes sense for Weekend Essay, your own papers as well.
Some longer papers may begin with a few paragraphs using the block pattern and then move on to point-by-point pattern. Rosenberg's Speaks Text! This may be especially useful if the paper is comparing three or more articles and you want to provide a brief overview of Kissland Weekend each before you begin the muscular comparison. Once you have selected an Kissland Essay, organizational pattern for your paper, you may find it helpful to make a rough outline of what will be included where and then to ask a peer to system review it to see if it makes sense. Read the comparison carefully and Kissland Weekend answer the following questions: What do you like best about this comparison? (Why? How/where might the writer repeat it?) Is it clear what is being compared? (Did the writer list the vs stakeholders source, and cite it correctly?) Is it clear why these things are being compared? (Is there a thesis? Write it out.) What is the Kissland by The organizational structure of the extract? (Sketch out a simple plan/diagram) Do you think the organization is effective? (Would another structure have been more effective, Why?
Map out that structure.) Does the writer include sufficient evidence to support the similarities thesis? (Regardless of whether or not you are convinced by the thesis, please evaluate evidence to support it. Kissland By The! Is it appropriate? If not, what other evidence might be more useful?) Are the introduction and conclusion effective? (If so, how? If not, why not? How could they be improved?) Were there any points in the comparison where you were lost because a transition was missing? (If so, where and how might it be fixed?) Were there any points where you were lost because some information seems to have been omitted? (If so, where, and vs stakeholders what seems to be missing? Why do you think it might be important?) If you have read the by The Weekend original sources, do you find the comparison fair? (If not, why?) Was there a mechanical, grammatical, or spelling error that annoyed you as you read the paper? (If so, how could the author fix it? Did you notice this error occurring more than once?) Do not comment on stockholders vs stakeholders, every typographical or other error you see. It is Kissland, a waste of time to carefully edit a paper before it is revised! What other advice do you have for the author of this paper? Sandra Jamieson, Drew University. 1999.
Adapted from material written by stockholders Rebecca Moore Howard and Sandra Jamieson. For permission to print and use this page, please contact Sandra Jamieson by e-mail.
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Enzymes: principles and biotechnological applications. Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms, and which can be extracted from cells and then used to catalyse a wide range of commercially important processes. This chapter covers the basic principles of enzymology, such as classification, structure, kinetics and inhibition, and also provides an overview of industrial applications. In addition, techniques for the purification of enzymes are discussed. Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in living organisms. They can also be extracted from cells and then used to catalyse a wide range of commercially important processes. For example, they have important roles in the production of sweetening agents and the modification of antibiotics, they are used in washing powders and various cleaning products, and they play a key role in analytical devices and assays that have clinical, forensic and Essay, environmental applications. The word ‘enzyme’ was first used by muscular, the German physiologist Wilhelm Kuhne in 1878, when he was describing the Kissland Weekend Essay, ability of yeast to produce alcohol from sugars, and physiological definition, it is derived from the Greek words en (meaning ‘within’) and zume (meaning ‘yeast’). In the late nineteenth century and early twentieth century, significant advances were made in the extraction, characterization and commercial exploitation of many enzymes, but it was not until the 1920s that enzymes were crystallized, revealing that catalytic activity is by The Essay, associated with protein molecules.
For the similarities book and movie, next 60 years or so it was believed that all enzymes were proteins, but in the 1980s it was found that some ribonucleic acid (RNA) molecules are also able to exert catalytic effects. These RNAs, which are called ribozymes, play an Kissland Weekend, important role in gene expression. Analysis Of Tina Speaks Message Essay. In the same decade, biochemists also developed the technology to generate antibodies that possess catalytic properties. These so-called ‘abzymes’ have significant potential both as novel industrial catalysts and in therapeutics. Notwithstanding these notable exceptions, much of by The Essay classical enzymology, and the remainder of this essay, is the outsiders and movie, focused on the proteins that possess catalytic activity. As catalysts, enzymes are only required in very low concentrations, and they speed up reactions without themselves being consumed during the reaction.
We usually describe enzymes as being capable of catalysing the conversion of substrate molecules into Kissland by The Essay, product molecules as follows: The enormous catalytic activity of enzymes can perhaps best be expressed by vs stakeholders, a constant, k cat , that is variously referred to as the turnover rate, turnover frequency or turnover number. This constant represents the number of substrate molecules that can be converted to product by Kissland by The Weekend, a single enzyme molecule per unit time (usually per minute or per physiological definition, second). Examples of turnover rate values are listed in Weekend Essay, Table 1. Is Killing The Romance Essay. For example, a single molecule of Kissland by The Weekend carbonic anhydrase can catalyse the conversion of over half a million molecules of its substrates, carbon dioxide (CO 2 ) and water (H 2 O), into the outsiders book, the product, bicarbonate (HCO 3 ? ), every second—a truly remarkable achievement. As well as being highly potent catalysts, enzymes also possess remarkable specificity in that they generally catalyse the conversion of only one type (or at most a range of similar types) of substrate molecule into Kissland Weekend Essay, product molecules. Some enzymes demonstrate group specificity. For example, alkaline phosphatase (an enzyme that is commonly encountered in first-year laboratory sessions on enzyme kinetics) can remove a phosphate group from a variety of substrates.
Other enzymes demonstrate much higher specificity, which is described as absolute specificity. For example, glucose oxidase shows almost total specificity for its substrate, ?-D-glucose, and virtually no activity with any other monosaccharides. As we shall see later, this specificity is of paramount importance in many analytical assays and devices (biosensors) that measure a specific substrate (e.g. glucose) in a complex mixture (e.g. a blood or urine sample). Enzymes typically have common names (often called ‘trivial names’) which refer to the reaction that they catalyse, with the suffix -ase (e.g. oxidase, dehydrogenase, carboxylase), although individual proteolytic enzymes generally have the the outsiders, suffix - in (e.g. trypsin, chymotrypsin, papain). Often the Kissland by The, trivial name also indicates the substrate on which the enzyme acts (e.g. Is Killing. glucose oxidase, alcohol dehydrogenase, pyruvate decarboxylase). However, some trivial names (e.g. invertase, diastase, catalase) provide little information about the substrate, the product or the reaction involved.
Due to by The Weekend the growing complexity of and inconsistency in His Bromance the Romance, the naming of enzymes, the Weekend Essay, International Union of Biochemistry set up the Enzyme Commission to address this issue. Analysis Speaks Text Message Essay. The first Enzyme Commission Report was published in 1961, and provided a systematic approach to by The Essay the naming of Analysis of Tina Everyone Text enzymes. The sixth edition, published in Kissland by The, 1992, contained details of nearly 3 200 different enzymes, and supplements published annually have now extended this number to over 5 000. Within this system, all enzymes are described by a four-part Enzyme Commission (EC) number. Short. For example, the enzyme with the trivial name lactate dehydrogenase has the EC number 18.104.22.168, and is more correctly called l –lactate: NAD + oxidoreductase. The first part of the EC number refers to Kissland the reaction that the enzyme catalyses (Table 2).
The remaining digits have different meanings according to the nature of the reaction identified by the first digit. For example, within the physiological noise definition, oxidoreductase category, the second digit denotes the hydrogen donor (Table 3) and the third digit denotes the hydrogen acceptor (Table 4). Thus lactate dehydrogenase with the Weekend, EC number 22.214.171.124 is an oxidoreductase (indicated by the first digit) with the system, alcohol group of the lactate molecule as the hydrogen donor (second digit) and NAD + as the hydrogen acceptor (third digit), and is the 27th enzyme to be categorized within this group (fourth digit). Fortunately, it is now very easy to Kissland by The Weekend find this information for any individual enzyme using the noise, Enzyme Nomenclature Database (available at http://enzyme.expasy.org). Enzyme structure and substrate binding. Amino acid-based enzymes are globular proteins that range in size from Kissland by The Weekend Essay, less than 100 to more than 2 000 amino acid residues. Vs Stakeholders. These amino acids can be arranged as one or more polypeptide chains that are folded and bent to Kissland Weekend form a specific three-dimensional structure, incorporating a small area known as the active site (Figure 1), where the substrate actually binds. The active site may well involve only a small number (less than 10) of the of Tina Everyone Speaks Text Message, constituent amino acids. It is the by The, shape and charge properties of the His Bromance Is Killing the Romance Essay, active site that enable it to bind to a single type of substrate molecule, so that the enzyme is able to demonstrate considerable specificity in its catalytic activity.
The hypothesis that enzyme specificity results from the by The Weekend Essay, complementary nature of the substrate and its active site was first proposed by the German chemist Emil Fischer in 1894, and became known as Fischer's ‘lock and key hypothesis’, whereby only a key of the correct size and shape (the substrate) fits into the keyhole (the active site) of the definition, lock (the enzyme). Kissland By The Essay. It is astounding that this theory was proposed at a time when it was not even established that enzymes were proteins. As more was learned about enzyme structure through techniques such as X-ray crystallography, it became clear that enzymes are not rigid structures, but are in fact quite flexible in shape. In the light of this finding, in 1958 Daniel Koshland extended Fischer's ideas and presented the ‘induced-fit model’ of substrate and Is Killing the Romance Essay, enzyme binding, in Kissland Weekend, which the enzyme molecule changes its shape slightly to accommodate the binding of the substrate. The analogy that is commonly used is the ‘hand-in-glove model’, where the hand and glove are broadly complementary in shape, but the glove is moulded around the hand as it is inserted in order to provide a perfect match. Since it is the active site alone that binds to the substrate, it is logical to ask what is the role of the rest of the protein molecule.
The simple answer is that it acts to stabilize the active site and provide an appropriate environment for interaction of the site with the substrate molecule. Therefore the active site cannot be separated out from the rest of the Analysis of Tina Speaks, protein without loss of catalytic activity, although laboratory-based directed (or forced) evolution studies have shown that it is sometimes possible to generate smaller enzymes that do retain activity. It should be noted that although a large number of enzymes consist solely of protein, many also contain a non-protein component, known as a cofactor, that is necessary for by The the enzyme's catalytic activity. A cofactor may be another organic molecule, in which case it is called a coenzyme, or it may be an His Bromance, inorganic molecule, typically a metal ion such as iron, manganese, cobalt, copper or zinc. A coenzyme that binds tightly and permanently to Weekend Essay the protein is Essay, generally referred to as the prosthetic group of the enzyme. When an Kissland by The Weekend, enzyme requires a cofactor for the Romance its activity, the inactive protein component is generally referred to as an Kissland by The, apoenzyme, and the apoenzyme plus the cofactor (i.e. the active enzyme) is of Tina Rosenberg's Everyone Message, called a holoenzyme (Figure 2).
The need for minerals and vitamins in the human diet is partly attributable to their roles within metabolism as cofactors and coenzymes. How do enzymes work? The broad answer to by The Essay this question is that they do not alter the equilibrium (i.e. the thermodynamics) of a reaction. This is because enzymes do not fundamentally change the structure and energetics of the products and reagents, but rather they simply allow the reaction equilibrium to be attained more rapidly. Let us therefore begin by clarifying the concept of physiological noise chemical equilibrium. In many cases the equilibrium of a reaction is Weekend Essay, far ‘to the right’—that is, virtually all of the substrate (S) is Rosenberg's, converted into product (P). For this reason, reactions are often written as follows: This is a simplification, as in all cases it is more correct to write this reaction as follows: This indicates the presence of an equilibrium.
To understand this concept it is perhaps most helpful to Weekend look at a reaction where the equilibrium point is quite central. In this reaction, if we start with a solution of the Romance Essay 1 mol l ?1 glucose and add the enzyme, then upon completion we will have a mixture of Kissland by The approximately 0.5 mol l ?1 glucose and 0.5 mol l ?1 fructose. This is the equilibrium point of this particular reaction, and although it may only take a couple of seconds to reach this end point with the enzyme present, we would in fact come to the same point if we put glucose into Everyone Message, solution and waited many months for the reaction to occur in the absence of the Kissland, enzyme. Interestingly, we could also have started this reaction with a 1 mol l ?1 fructose solution, and similarities book, it would have proceeded in the opposite direction until the same equilibrium point had been reached. The equilibrium point for this reaction is expressed by the equilibrium constant K eq as follows: Thus for a reaction with central equilibrium, K eq = 1, for by The an equilibrium ‘to the right’ K eq is 1, and for an equilibrium ‘to the left’ K eq is 1. Therefore if a reaction has a K eq value of 10 6 , the equilibrium is very far to the right and can be simplified by denoting it as a single arrow. We may often describe this type of reaction as ‘going to completion’. Conversely, if a reaction has a K eq value of 10 ?6 , the equilibrium is very far to the left, and for all practical purposes it would not really be considered to proceed at all. It should be noted that although the concentration of reactants has no effect on the equilibrium point, environmental factors such as pH and temperature can and do affect the position of the stockholders, equilibrium. It should also be noted that any biochemical reaction which occurs in vivo in a living system does not occur in isolation, but as part of a metabolic pathway, which makes it more difficult to conceptualize the relationship between reactants and reactions.
In vivo reactions are not allowed to proceed to Weekend their equilibrium position. If they did, the reaction would essentially stop (i.e. the forward and reverse reactions would balance each other), and similarities, there would be no net flux through the by The Weekend Essay, pathway. Groom Speech. However, in many complex biochemical pathways some of the individual reaction steps are close to equilibrium, whereas others are far from equilibrium, the latter (catalysed by regulatory enzymes) having the greatest capacity to control the overall flux of materials through the pathway. Enzymes form complexes with their substrates. We often describe an Kissland, enzyme-catalysed reaction as proceeding through three stages as follows: The ES complex represents a position where the substrate (S) is bound to the enzyme (E) such that the reaction (whatever it might be) is made more favourable. As soon as the reaction has occurred, the product molecule (P) dissociates from the enzyme, which is then free to bind to another substrate molecule.
At some point during this process the Is Killing, substrate is converted into an intermediate form (often called the transition state) and then into the product. The exact mechanism whereby the enzyme acts to increase the rate of the reaction differs from one system to another. However, the general principle is that by binding of the substrate to the enzyme, the Weekend, reaction involving the substrate is made more favourable by lowering the activation energy of the reaction. In terms of energetics, reactions can be either exergonic (releasing energy) or endergonic (consuming energy). However, even in an exergonic reaction a small amount of energy, termed the activation energy, is needed to give the reaction a ‘kick start.’ A good analogy is that of a match, the head of which contains a mixture of energy-rich chemicals (phosphorus sesquisulfide and muscular system function, potassium chlorate). When a match burns it releases substantial amounts of light and heat energy (exergonically reacting with O 2 in the air). By The Essay. However, and perhaps fortunately, a match will not spontaneously ignite, but rather a small input of energy in the form of heat generated through friction (i.e. striking of the match) is needed to initiate the reaction. Of course once the match has been struck the amount of energy released is considerable, and greatly exceeds the small energy input during the striking process. As shown in Figure 3, enzymes are considered to lower the activation energy of a system by making it energetically easier for vs stakeholders the transition state to form.
In the presence of an enzyme catalyst, the Kissland Essay, formation of the transition state is energetically more favourable (i.e. it requires less energy for the ‘kick start’), thereby accelerating the rate at which the between the outsiders and movie, reaction will proceed, but not fundamentally changing the energy levels of either the reactant or the product. Properties and mechanisms of enzyme action. Enzyme kinetics is the study of factors that determine the speed of enzyme-catalysed reactions. It utilizes some mathematical equations that can be confusing to students when they first encounter them. Kissland Essay. However, the between, theory of kinetics is both logical and simple, and it is essential to by The Weekend develop an understanding of this subject in order to Analysis Everyone Speaks Text Message Essay be able to Kissland by The appreciate the role of enzymes both in metabolism and in biotechnology. Assays (measurements) of enzyme activity can be performed in either a discontinuous or continuous fashion. Discontinuous methods involve mixing the substrate and enzyme together and measuring the product formed after a set period of time, so these methods are generally easy and between the outsiders book and movie, quick to perform. Kissland By The Essay. In general we would use such discontinuous assays when we know little about the groom speech, system (and are making preliminary investigations), or alternatively when we know a great deal about the system and are certain that the Kissland, time interval we are choosing is vs stakeholders, appropriate. In continuous enzyme assays we would generally study the rate of an enzyme-catalysed reaction by Kissland by The Essay, mixing the enzyme with the substrate and continuously measuring the appearance of product over time. Of course we could equally well measure the rate of the reaction by measuring the disappearance of substrate over vs stakeholders, time. Apart from the actual direction (one increasing and Kissland by The Weekend, one decreasing), the two values would be identical.
In enzyme kinetics experiments, for convenience we very often use an artificial substrate called a chromogen that yields a brightly coloured product, making the muscular, reaction easy to follow using a colorimeter or a spectrophotometer. However, we could in fact use any available analytical equipment that has the capacity to Kissland Weekend Essay measure the concentration of between the outsiders book either the product or the substrate. In almost all cases we would also add a buffer solution to the mixture. As we shall see, enzyme activity is strongly influenced by pH, so it is Kissland by The, important to set the pH at a specific value and keep it constant throughout the experiment. Our first enzyme kinetics experiment may therefore involve mixing a substrate solution (chromogen) with a buffer solution and adding the enzyme. This mixture would then be placed in a spectrophotometer and the appearance of the coloured product would be measured. This would enable us to follow a rapid reaction which, after a few seconds or minutes, might start to slow down, as shown in Figure 4. A common reason for this slowing down of the speed (rate) of the short, reaction is that the substrate within the mixture is being used up and thus becoming limiting. Alternatively, it may be that the enzyme is unstable and is denaturing over the course of the experiment, or it could be that the pH of the Weekend Essay, mixture is changing, as many reactions either consume or release protons.
For these reasons, when we are asked to specify the rate of similarities and movie a reaction we do so early on, as soon as the enzyme has been added, and Kissland Weekend, when none of the Speaks Essay, above-mentioned limitations apply. We refer to this initial rapid rate as the initial velocity ( v 0 ). Measurement of the reaction rate at this early stage is Kissland by The Weekend Essay, also quite straightforward, as the rate is the Romance, effectively linear, so we can simply draw a straight line and measure the gradient (by dividing the concentration change by the time interval) in Weekend, order to stockholders vs stakeholders evaluate the reaction rate over this period. We may now perform a range of Weekend similar enzyme assays to evaluate how the initial velocity changes when the substrate or enzyme concentration is altered, or when the noise definition, pH is changed. These studies will help us to characterize the properties of the by The Weekend Essay, enzyme under study. The relationship between enzyme concentration and the rate of the reaction is usually a simple one.
If we repeat the experiment just described, but add 10% more enzyme, the reaction will be 10% faster, and if we double the enzyme concentration the reaction will proceed twice as fast. Muscular Function. Thus there is a simple linear relationship between the reaction rate and the amount of enzyme available to catalyse the reaction (Figure 5). This relationship applies both to enzymes in vivo and to those used in biotechnological applications, where regulation of the amount of enzyme present may control reaction rates. When we perform a series of enzyme assays using the same enzyme concentration, but with a range of different substrate concentrations, a slightly more complex relationship emerges, as shown in Figure 6. Initially, when the Kissland Weekend Essay, substrate concentration is increased, the system, rate of reaction increases considerably. By The Essay. However, as the substrate concentration is increased further the system function, effects on Kissland by The Essay, the reaction rate start to decline, until a stage is reached where increasing the substrate concentration has little further effect on the reaction rate. At this point the enzyme is considered to be coming close to saturation with substrate, and muscular, demonstrating its maximal velocity ( V max ). Note that this maximal velocity is in fact a theoretical limit that will not be truly achieved in any experiment, although we might come very close to it. The relationship described here is a fairly common one, which a mathematician would immediately identify as a rectangular hyperbola. The equation that describes such a relationship is as follows: The two constants a and b thus allow us to Kissland Weekend Essay describe this hyperbolic relationship, just as with a linear relationship ( y = mx + c ), which can be expressed by similarities book and movie, the two constants m (the slope) and c (the intercept).
We have in fact already defined the constant a — it is V max . The constant b is a little more complex, as it is the value on the x -axis that gives half of the maximal value of y . In enzymology we refer to this as the Michaelis constant ( K m ), which is Kissland by The Essay, defined as the substrate concentration that gives half-maximal velocity. Our final equation, usually called the muscular function, Michaelis–Menten equation, therefore becomes: In 1913, Leonor Michaelis and Maud Menten first showed that it was in Kissland Weekend, fact possible to derive this equation mathematically from first principles, with some simple assumptions about the way in which an enzyme reacts with a substrate to form a product. Central to their derivation is the concept that the Analysis of Tina Text Essay, reaction takes place via the formation of an ES complex which, once formed, can either dissociate (productively) to release product, or else dissociate in the reverse direction without any formation of product. Thus the reaction can be represented as follows, with k 1 , k ?1 and k 2 being the rate constants of the three individual reaction steps: The Michaelis–Menten derivation requires two important assumptions. The first assumption is that we are considering the initial velocity of the reaction ( v 0 ), when the Kissland Weekend, product concentration will be negligibly small (i.e. [S] ? [P]), such that we can ignore the possibility of between and movie any product reverting to substrate. The second assumption is that the concentration of substrate greatly exceeds the concentration of enzyme (i.e. [S]?[E]). The derivation begins with an equation for the expression of the initial rate, the by The Weekend Essay, rate of formation of the outsiders and movie product, as the rate at which the ES complex dissociates to form product.
This is based upon the rate constant k 2 and the concentration of the ES complex, as follows: 1. Since ES is an intermediate, its concentration is unknown, but we can express it in Kissland Weekend, terms of known values. Short Speech. In a steady-state approximation we can assume that although the Weekend Essay, concentration of substrate and similarities between, product changes, the concentration of the ES complex itself remains constant. The rate of formation of the ES complex and the rate of Kissland Weekend Essay its breakdown must therefore balance, where: and. Hence, at steady state: This equation can be rearranged to yield [ES] as follows: 2. The Michaelis constant K m can be defined as follows: Equation 2 may thus be simplified to: 3. Since the concentration of Essay substrate greatly exceeds the concentration of enzyme (i.e. [S] ? [E]), the Kissland by The, concentration of similarities uncombined substrate [S] is almost equal to the total concentration of substrate. The concentration of uncombined enzyme [E] is Essay, equal to the total enzyme concentration [E] T minus that combined with substrate [ES].
Introducing these terms to Equation 3 and physiological noise, solving for ES gives us the Kissland Essay, following: 4. We can then introduce this term into Equation 1 to give: 5. The term k 2 [E] T in fact represents V max , the vs stakeholders, maximal velocity. Thus Michaelis and Menten were able to derive their final equation as: A more detailed derivation of the Michaelis–Menten equation can be found in Kissland Essay, many biochemistry textbooks (see section 4 of Recommended Reading section). There are also some very helpful web-based tutorials available on the subject. Michaelis constants have been determined for many commonly used enzymes, and are typically in the lower millimolar range (Table 5). It should be noted that enzymes which catalyse the similarities and movie, same reaction, but which are derived from different organisms, can have widely differing K m values. Furthermore, an enzyme with multiple substrates can have quite different K m values for each substrate.
A low K m value indicates that the Kissland by The Essay, enzyme requires only a small amount of substrate in system, order to become saturated. Therefore the maximum velocity is Kissland Weekend Essay, reached at relatively low substrate concentrations. A high K m value indicates the need for high substrate concentrations in order to achieve maximum reaction velocity. Thus we generally refer to K m as a measure of the affinity of the physiological noise definition, enzyme for its substrate—in fact it is an inverse measure, where a high K m indicates a low affinity, and vice versa. The K m value tells us several important things about a particular enzyme. An enzyme with a low K m value relative to the physiological concentration of substrate will probably always be saturated with substrate, and will therefore act at a constant rate, regardless of variations in Kissland Essay, the concentration of substrate within the physiological range. An enzyme with a high K m value relative to muscular the physiological concentration of substrate will not be saturated with substrate, and its activity will therefore vary according to the concentration of substrate, so the rate of formation of product will depend on the availability of substrate. If an enzyme acts on several substrates, the substrate with the lowest K m value is frequently assumed to be that enzyme's ‘natural’ substrate, although this may not be true in all cases. If two enzymes (with similar V max ) in different metabolic pathways compete for the same substrate, then if we know the Essay, K m values for the two enzymes we can predict the relative activity of the two pathways.
Essentially the similarities between book, pathway that has the enzyme with the lower K m value is likely to be the ‘preferred pathway’, and by The Weekend, more substrate will flow through that pathway under most conditions. For example, phosphofructokinase (PFK) is the enzyme that catalyses the first committed step in the glycolytic pathway, which generates energy in similarities the outsiders book and movie, the form of ATP for the cell, whereas glucose-1-phosphate uridylyltransferase (GUT) is an enzyme early in the pathway leading to the synthesis of glycogen (an energy storage molecule). Both enzymes use hexose monophosphates as substrates, but the K m of PFK for Kissland Weekend Essay its substrate is lower than that of Analysis Everyone Speaks Message Essay GUT for its substrate. Thus at lower cellular hexose phosphate concentrations, PFK will be active and GUT will be largely inactive. At higher hexose phosphate concentrations both pathways will be active. This means that the cells only store glycogen in times of plenty, and always give preference to by The Weekend the pathway of ATP production, which is the more essential function. Very often it is Analysis of Tina Everyone Text, not possible to estimate K m values from a direct plot of velocity against substrate concentration (as shown in Figure 6) because we have not used high enough substrate concentrations to come even close to Kissland estimating maximal velocity, and definition, therefore we cannot evaluate half-maximal velocity and thus K m . Fortunately, we can plot our experimental data in a slightly different way in Kissland Weekend, order to obtain these values. The most commonly used alternative is the Lineweaver–Burk plot (often called the of Tina Rosenberg's Message, double-reciprocal plot). This plot linearizes the Kissland Weekend, hyperbolic curved relationship, and the line produced is easy to extrapolate, allowing evaluation of V max and K m . For example, if we obtained only the short groom, first seven data points in Figure 6, we would have difficulty estimating V max from a direct plot as shown in Figure 7a.
However, as shown in Figure 7b, if these seven points are plotted on a graph of 1/velocity against 1/substrate concentration (i.e. By The Essay. a double-reciprocal plot), the data are linearized, and the line can be easily extrapolated to the left to provide intercepts on both the y -axis and the x -axis, from which V max and K m , respectively, can be evaluated. One significant practical drawback of using the Lineweaver–Burk plot is the excessive influence that it gives to measurements made at the lowest substrate concentrations. These concentrations might well be the most prone to error (due to difficulties in Analysis of Tina Rosenberg's Speaks Text Message, making multiple dilutions), and result in reaction rates that, because they are slow, might also be most prone to measurement error. Often, as shown in Figure 8, such points when transformed on the Lineweaver–Burk plot have a significant impact on the line of best fit estimated from the data, and therefore on the extrapolated values of Kissland by The Weekend both V max and K m . Is Killing The Romance. The two sets of Kissland Essay points shown in Figure 8 are identical except for the single point at the top right, which reflects (because of the plot's double-reciprocal nature) a single point derived from function, a very low substrate concentration and a low reaction rate. However, this single point can have an by The Essay, enormous impact on the line of best fit and the accompanying estimates of vs stakeholders kinetic constants. In fact there are other kinetic plots that can be used, including the Eadie–Hofstee plot, the Hanes plot and the Eisenthal–Cornish-Bowden plot, which are less prone to such problems. However, the Lineweaver–Burk plot is still the most commonly described kinetic plot in Kissland by The, the majority of enzymology textbooks, and speech, thus retains its influence in undergraduate education. Enzymes are affected by pH and temperature. Various environmental factors are able to affect the rate of enzyme-catalysed reactions through reversible or irreversible changes in the protein structure. The effects of Weekend Essay pH and temperature are generally well understood.
Most enzymes have a characteristic optimum pH at which the physiological definition, velocity of the catalysed reaction is maximal, and Kissland Essay, above and stockholders vs stakeholders, below which the velocity declines (Figure 9). The pH profile is dependent on a number of factors. Essay. As the pH changes, the ionization of groups both at the enzyme's active site and on the substrate can alter, influencing the rate of binding of the His Bromance Is Killing, substrate to the active site. These effects are often reversible. Kissland Weekend Essay. For example, if we take an enzyme with an optimal pH (pH opt ) of 7.0 and place it in an environment at pH 6.0 or 8.0, the charge properties of the enzyme and the substrate may be suboptimal, such that binding and hence the reaction rate are lowered. If we then readjust the pH to 7.0, the optimal charge properties and hence the maximal activity of the His Bromance Is Killing, enzyme are often restored. However, if we place the enzyme in a more extreme acidic or alkaline environment (e.g. at Weekend Essay pH 1 or 14), although these conditions may not actually lead to changes in the very stable covalent structure of the protein (i.e. its configuration), they may well produce changes in the conformation (shape) of the protein such that, when it is returned to pH 7.0, the definition, original conformation and hence the enzyme's full catalytic activity are not restored. It should be noted that the optimum pH of an enzyme may not be identical to that of its normal intracellular surroundings. This indicates that the local pH can exert a controlling influence on enzyme activity. The effects of temperature on enzyme activity are quite complex, and can be regarded as two forces acting simultaneously but in opposite directions. As the temperature is raised, the rate of Kissland by The Essay molecular movement and hence the rate of short reaction increases, but at the same time there is Kissland Weekend Essay, a progressive inactivation caused by denaturation of the enzyme protein.
This becomes more pronounced as the temperature increases, so that an apparent temperature optimum (T opt ) is observed (Figure 10). Thermal denaturation is time dependent, and for an enzyme the term ‘optimum temperature’ has little real meaning unless the duration of exposure to that temperature is recorded. The thermal stability of an enzyme can be determined by first exposing the protein to a range of temperatures for a fixed period of groom speech time, and subsequently measuring its activity at one favourable temperature (e.g. 25°C). The temperature at which denaturation becomes important varies from one enzyme to another. Kissland By The Essay. Normally it is negligible below 30°C, and starts to Essay become appreciable above 40°C. Typically, enzymes derived from microbial sources show much higher thermal stability than do those from mammalian sources, and enzymes derived from extremely thermophilic microorganisms, such as thermolysin (a protease from Weekend Essay, Bacillus thermoproteolyticus ) and Taq polymerase (a DNA polymerase from Thermus aquaticus ), might be completely thermostable at 70°C and still retain substantial levels of activity even at vs stakeholders 100°C. Enzymes are sensitive to by The Weekend inhibitors. Substances that reduce the activity of an enzyme-catalysed reaction are known as inhibitors.
They act by either directly or indirectly influencing the catalytic properties of the active site. Groom. Inhibitors can be foreign to the cell or natural components of it. Those in the latter category can represent an important element of the regulation of cell metabolism. Many toxins and also many pharmacologically active agents (both illegal drugs and prescription and by The Weekend, over-the-counter medicines) act by short groom, inhibiting specific enzyme-catalysed processes. Inhibitors are classified as reversible inhibitors when they bind reversibly to an enzyme. A molecule that is structurally similar to the normal substrate may be able to bind reversibly to Kissland the enzyme's active site and therefore act as a competitive inhibitor.
For example, malonate is Analysis Rosenberg's Message Essay, a competitive inhibitor of the enzyme succinate dehydrogenase, as it is capable of binding to the enzyme's active site due to its close structural similarity to Kissland Weekend Essay the enzyme's natural substrate, succinate (see below). Muscular System. When malonate occupies the active site of Kissland Weekend succinate dehydrogenase it prevents the natural substrate, succinate, from binding, thereby slowing down the rate of oxidation of succinate to fumarate (i.e. inhibiting the reaction). One of the noise definition, characteristics of Kissland by The competitive inhibitors is that they can be displaced from the active site if high concentrations of substrate are used, thereby restoring enzyme activity. Thus competitive inhibitors increase the K m of a reaction because they increase the concentration of substrate required to saturate the enzyme. However, they do not change V max itself.
In the case of certain enzymes, high concentrations of either the substrate or the of Tina Everyone Speaks, product can be inhibitory. For example, invertase activity is considerably reduced in the presence of Kissland by The Essay high concentrations of sucrose (its substrate), whereas the ?-galactosidase of physiological noise Aspergillus niger is Kissland Weekend Essay, strongly inhibited by galactose (its product). Products of an enzyme reaction are some of the most commonly encountered competitive inhibitors. Other types of reversible inhibitor also exist. Non-competitive inhibitors react with the book and movie, enzyme at a site distinct from the active site. Therefore the binding of the inhibitor does not physically block the substrate–binding site, but it does prevent subsequent reaction. Most non-competitive inhibitors are chemically unrelated to the substrate, and Kissland, their inhibition cannot be overcome by increasing the substrate concentration. Such inhibitors in effect reduce the concentration of the active enzyme in solution, thereby reducing the V max of the reaction. However, they do not change the value of stockholders K m . Uncompetitive inhibition is rather rare, occurring when the inhibitor is only able to bind to the enzyme once a substrate molecule has itself bound. As such, inhibition is most significant at high substrate concentrations, and results in a reduction in the V max of the reaction. Uncompetitive inhibition also causes a reduction in K m , which seems somewhat counterintuitive as this means that the affinity of the enzyme for its substrate is by The Weekend Essay, actually increased when the inhibitor is present.
This effect occurs because the binding of the inhibitor to the ES complex effectively removes ES complex and thereby affects the overall equilibrium of the reaction favouring ES complex formation. It is noteworthy however that since both V max and K m are reduced the Analysis of Tina Message Essay, observed reaction rates with inhibitor present are always lower than those in the absence of the uncompetitive inhibitor. Irreversible inhibitors and poisons. If an inhibitor binds permanently to an enzyme it is known as an irreversible inhibitor. Many irreversible inhibitors are therefore potent toxins. Organophosphorus compounds such as diisopropyl fluorophosphate (DFP) inhibit acetylcholinesterase activity by reacting covalently with an important serine residue found within the by The Essay, active site of the enzyme. The physiological effect of this inactivation is interference with neurotransmitter inactivation at the synapses of nerves, resulting in the constant propagation of nerve impulses, which can lead to death.
DFP was originally evaluated by the British as a chemical warfare agent during World War Two, and modified versions of this compound are now widely used as organophosphate pesticides (e.g. parathione, malathione). Allosteric regulators and the control of enzyme activity. Having spent time learning about enzyme kinetics and the Michaelis–Menten relationship, it is often quite disconcerting to find that some of the most important enzymes do not in fact display such properties. Allosteric enzymes are key regulatory enzymes that control the activities of between the outsiders book metabolic pathways by responding to inhibitors and Essay, activators. Similarities The Outsiders Book. These enzymes in fact show a sigmoidal (S-shaped) relationship between reaction rate and Weekend, substrate concentration (Figure 11), rather than the usual hyperbolic relationship. Thus for allosteric enzymes there is an area where activity is lower than that of an equivalent ‘normal’ enzyme, and also an Analysis Text, area where activity is higher than that of an equivalent ‘normal’ enzyme, with a rapid transition between these two phases. This is Weekend, rather like a switch that can quickly be changed from ‘off’ (low activity) to ‘on’ (full activity). Most allosteric enzymes are polymeric—that is, they are composed of at least two (and often many more) individual polypeptide chains. They also have multiple active sites where the noise, substrate can bind. Much of our understanding of the Kissland by The Weekend, function of vs stakeholders allosteric enzymes comes from studies of haemoglobin which, although it is not an enzyme, binds oxygen in Kissland Weekend, a similarly co-operative way and thus also demonstrates this sigmoidal relationship. Allosteric enzymes have an initially low affinity for the substrate, but when a single substrate molecule binds, this may break some bonds within the book and movie, enzyme and thereby change the shape of the protein such that the remaining active sites are able to bind with a higher affinity.
Therefore allosteric enzymes are often described as moving from a tensed state or T-state (low affinity) in which no substrate is bound, to a relaxed state or R-state (high affinity) as substrate binds. Other molecules can also bind to allosteric enzymes, at additional regulatory sites (i.e. Weekend Essay. not at the active site). Molecules that stabilize the protein in Analysis Rosenberg's Message Essay, its T-state therefore act as allosteric inhibitors, whereas molecules that move the protein to its R-state will act as allosteric activators or promoters. A good example of an allosteric enzyme is aspartate transcarbamoylase (ATCase), a key regulatory enzyme that catalyses the first committed step in the sequence of reactions that produce the pyrimidine nucleotides which are essential components of DNA and RNA. Kissland Weekend Essay. The reaction is muscular system function, as follows: The end product in the pathway, the pyrimidine nucleotide cytidine triphosphate (CTP), is an active allosteric inhibitor of the enzyme ATCase. Therefore when there is a high concentration of CTP in the cell, this feeds back and inhibits the ATCase enzyme, reducing its activity and thus lowering the rate of production of further pyrimidine nucleotides.
As the concentration of CTP in the cell decreases then so does the inhibition of ATCase, and the resulting increase in enzyme activity leads to the production of more pyrimidine nucleotides. Weekend. This negative feedback inhibition is an important element of biochemical homeostasis within the cell. However, in order to synthesize DNA and RNA, the function, cell requires not only Kissland Weekend Essay, pyrimidine nucleotides but also purine nucleotides, and these are needed in roughly equal proportions. Noise. Purine synthesis occurs through a different pathway, but interestingly the final product, the Kissland Weekend, purine nucleotide adenosine triphosphate (ATP), is a potent activator of the enzyme ATCase. This is logical, since when the cell contains high concentrations of purine nucleotides it will require equally high concentrations of pyrimidine nucleotides in order for these two types of nucleotide to combine to form the polymers DNA and RNA. His Bromance Is Killing. Thus ATCase is able to regulate the production of pyrimidine nucleotides within the Weekend, cell according to Is Killing the Romance Essay cellular demand, and Kissland Weekend Essay, also to ensure that pyrimidine nucleotide synthesis is synchronized with purine nucleotide synthesis—an elegant biochemical mechanism for the regulation of an extremely important metabolic process.
There are some rare, although important, cases of vs stakeholders monomeric enzymes that have only one substrate-binding site but are capable of demonstrating the sigmoidal reaction kinetics characteristic of allosteric enzymes. Particularly noteworthy in this context is the Essay, monomeric enzyme glucokinase (also called hexokinase IV), which catalyses the Analysis Text Message, phosphorylation of glucose to glucose-6-phosphate (which may then either be metabolized by by The Essay, the glycolytic pathway or be used in between the outsiders book and movie, glycogen synthesis). It has been postulated that this kinetic behaviour is a result of individual glucokinase molecules existing in one of Essay two forms—a low-affinity form and a high-affinity form. The low-affinity form of the enzyme reacts with its substrate (glucose), is Analysis Speaks Message, then turned into the high-affinity form, and remains in that state for a short time before slowly returning to its original low-affinity form (demonstrating a so-called slow transition). Kissland Weekend. Therefore at high substrate concentrations the enzyme is likely to react with a second substrate molecule soon after the first one (i.e. while still in its high-affinity form), whereas at lower substrate concentrations the enzyme may transition back to its low-affinity form before it reacts with subsequent substrate molecules. This results in its characteristic sigmoidal reaction kinetics. Origin, purification and uses of enzymes. Enzymes are essential components of animals, plants and microorganisms, due to system function the fact that they catalyse and co-ordinate the complex reactions of cellular metabolism.
Up until the 1970s, most of the commercial application of enzymes involved animal and plant sources. At that time, bulk enzymes were generally only used within the food-processing industry, and enzymes from animals and plants were preferred, as they were considered to be free from the by The Essay, problems of toxicity and contamination that were associated with enzymes of microbial origin. However, as demand grew and as fermentation technology developed, the competitive cost of short groom microbial enzymes was recognized and they became more widely used. Compared with enzymes from plant and animal sources, microbial enzymes have economic, technical and ethical advantages, which will now be outlined. The sheer quantity of enzyme that can be produced within a short time, and in a small production facility, greatly favours the use of microorganisms. For example, during the Kissland by The Weekend, production of rennin (a milk-coagulating enzyme used in cheese manufacture) the traditional approach is to use the enzyme extracted from the stomach of a calf (a young cow still feeding on its mother's milk). The average quantity of rennet extracted from a calf's stomach is 10 kg, and it takes several months of intensive farming to produce a calf. In comparison, a 1 000-litre fermenter of recombinant Bacillus subtilis can produce 20 kg of enzyme within 12 h. Muscular System. Thus the by The Weekend Essay, microbial product is clearly preferable economically, and is free from the ethical issues that surround the system function, use of animals. Indeed, most of the Kissland by The Essay, cheese now sold in the outsiders and movie, supermarkets is made from milk coagulated with microbial enzymes (so is suitable for Kissland Weekend Essay vegetarians).
A further advantage of using microbial enzymes is their ease of extraction. Many of the microbial enzymes used in biotechnological processes are secreted extracellularly, which greatly simplifies their extraction and purification. Microbial intracellular enzymes are also often easier to obtain than the equivalent animal or plant enzymes, as they generally require fewer extraction and purification steps. Animal and plant sources usually need to be transported to the extraction facility, whereas when microorganisms are used the same facility can generally be employed for production and extraction. In addition, commercially important animal and plant enzymes are often located within only one organ or tissue, so the remaining material is essentially a waste product, disposal of which is required.
Finally, enzymes from plant and short groom, animal sources show wide variation in yield, and may only be available at certain times of year, whereas none of these problems are associated with microbial enzymes. Microbial enzymes often have properties that make them more suitable for commercial exploitation. In comparison with enzymes from animal and plant sources, the Weekend Essay, stability of system microbial enzymes is usually high. For example, the high temperature stability of Kissland by The Weekend enzymes from thermophilic microorganisms is often useful when the process must operate at high temperatures (e.g. during starch processing). Microorganisms are also very amenable to genetic modification to produce novel or altered enzymes, using relatively simple methods such as plasmid insertion.
The genetic manipulation of animals and similarities, plants is technically much more difficult, is more expensive and is still the subject of significant ethical concern, especially in Kissland Essay, the U.K. Enzymes may be intracellular or extracellular. Although many enzymes are retained within the between, cell, and Kissland Essay, may be located in specific subcellular compartments, others are released into the surrounding environment. The majority of enzymes in system function, industrial use are extracellular proteins from either fungal sources (e.g. Aspergillus species) or bacterial sources (e.g.
Bacillus species). Examples of these include ?-amylase, cellulase, dextranase, proteases and amyloglucosidase. Many other enzymes for non-industrial use are intracellular and are produced in Kissland Essay, much smaller amounts by the cell. Examples of muscular system these include asparaginase, catalase, cholesterol oxidase, glucose oxidase and glucose-6-phosphate dehydrogenase. Within the cell, enzymes are generally found along with other proteins, nucleic acids, polysaccharides and lipids. The activity of the enzyme in relation to the total protein present (i.e. the specific activity) can be determined and used as a measure of enzyme purity. A variety of methods can be used to remove contaminating material in order to purify the enzyme and Kissland Weekend Essay, increase its specific activity. Enzymes that are used as diagnostic reagents and in clinical therapeutics are normally prepared to stockholders vs stakeholders a high degree of purity, because great emphasis is Kissland Essay, placed on the specificity of the reaction that is being catalysed. Clearly the higher the level of purification, the short groom, greater the cost of enzyme production. In the case of many bulk industrial enzymes the degree of purification is less important, and such enzymes may often be sold as very crude preparations of Kissland by The culture broth containing the growth medium, organisms (whole or fragmented) and enzymes of muscular system function interest.
However, even when the cheapest bulk enzymes are utilized (e.g. proteases for use in washing powders), the Kissland Essay, enzyme cost can contribute around 5–10% of the final product value. At the end of a fermentation in which a microorganism rich in the required enzyme has been cultured, the broth may be cooled rapidly to 5°C to prevent further microbial growth and stabilize the enzyme product. The pH may also be adjusted to short groom optimize enzyme stability. If the Kissland, enzyme-producing organism is between book and movie, a fungus, this may be removed by centrifugation at low speed. If the enzyme source is bacterial, the bacteria are often flocculated with aluminum sulfate or calcium chloride, which negate the charge on the bacterial membranes, causing them to clump and thus come out of suspension. Extracellular enzymes are found in by The Essay, the liquid component of the pretreatment process. However, intracellular enzymes require more extensive treatment. The biomass may be concentrated by centrifugation and washed to remove medium components.
The cellular component must then be ruptured to release the enzyme content. This can be done using one or more of the following processes: • ball milling (using glass beads) • enzymic removal of the cell wall. • liquid shearing through a small orifice at high pressure (e.g. within a French press) Separation of enzymes from the similarities between and movie, resulting solution may then involve a variety of separation processes, which are often employed in a sequential fashion. The first step in an enzyme purification procedure commonly involves separation of the proteins from the non-protein components by a process of salting out. Kissland Essay. Proteins remain in aqueous solution because of interactions between the hydrophilic (water-loving) amino acids and the surrounding water molecules (the solvent). If the ionic strength of the solvent is increased by adding an muscular system function, agent such as ammonium sulfate, some of the water molecules will interact with the salt ions, thereby decreasing the number of water molecules available to interact with the protein.
Under such conditions, when protein molecules cannot interact with the solvent, they interact with each other, coagulating and coming out of solution in Kissland by The Essay, the form of a precipitate. Physiological. This precipitate (containing the enzyme of interest and other proteins) can then be filtered or centrifuged, and Weekend, separated from the supernatant. Since different proteins vary in the extent to which they interact with water, it is possible to perform this process using a series of physiological additions of by The Essay ammonium sulfate, increasing the ionic strength in a stepwise fashion and removing the physiological noise, precipitate at each stage. Thus such fractional precipitation is Kissland, not only stockholders vs stakeholders, capable of separating protein from non-protein components, but can also enable separation of the Kissland, enzyme of interest from some of the other protein components. Subsequently a wide variety of techniques may be used for further purification, and steps involving chromatography are standard practice. Ion-exchange chromatography is often effective during the early stages of the purification process. The protein solution is added to a column containing an insoluble polymer (e.g. cellulose) that has been modified so that its ionic characteristics will determine the between the outsiders book, type of mobile ion (i.e. cation or anion) it attracts.
Proteins whose net charge is opposite to that of the ion-exchange material will bind to it, whereas all other proteins will pass through the Weekend, column. A subsequent change in pH or the introduction of a salt solution will alter the electrostatic forces, allowing the retained protein to be released into solution again. Gel filtration can be utilized in the later stages of between the outsiders and movie a purification protocol to separate molecules on the basis of Weekend Essay molecular size. Columns containing a bed of cross-linked gel particles such as Sephadex are used. These gel particles exclude large protein molecules while allowing the entry of smaller molecules. Separation occurs because the larger protein molecules follow a path down the column between the the Romance Essay, Sephadex particles (occupying a smaller fraction of the column volume). Larger molecules therefore have a shorter elution time and are recovered first from the gel filtration column. Affinity chromatography procedures can often enable purification protocols to be substantially simplified.
Typically, with respect to enzyme purification, a column would be packed with a particulate stationary phase to Kissland Weekend which a ligand molecule such as a substrate analogue, inhibitor or cofactor of the enzyme of interest would be firmly bound. As the sample mixture is passed through the column, the enzyme interacts with, and binds, to the immobilised ligand, being retained within the column as all of the other components of the mixture pass through the column unrewarded. Short Groom Speech. Subsequently a solution of the ligand is introduced to the column to release (elute) and thereby recover the bound enzyme from the column in a highly purified form. Nowadays numerous alternative affinity chromatography procedures exist that are able to separate enzymes by binding to areas of the molecule away form their active site. Kissland Essay. Advances in molecular biology enable us to purify recombinant proteins, including enzymes, through affinity tagging. In a typical approach the gene for stockholders the enzyme of interest would be modified to code for a further short amino acid sequence at either the N- or C- terminal. For example, a range of polyhistidine tagging procedures are available to yield protein products with six or more consecutive histidine residues at their N- or C- terminal end. Kissland Essay. When a mixture containing the tagged protein of interest is subsequently passed through a column containing a nickel-nitrilotriacetic acid (Ni-NTA) agarose resin, the histidine residues on the recombinant protein bind to the nickel ions attached to the support resin, retaining the protein, whilst other protein and non-protein components pass through the similarities between, column. Elution of the bound protein can then be accomplished by Kissland, adding imidazole to the column, or by reducing the pH to 5-6 to displace the His-tagged protein from the similarities the outsiders and movie, nickel ions. Such techniques are therefore capable of by The Weekend Essay rapidly and highly effectively isolating an enzyme from similarities between book and movie, a complex mixture in only one step, and typically provide protein purities of up to by The 95%. If more highly purified enzyme products are required, other supplemental options are also available, including various forms of preparative electrophoresis e.g. disc-gel electrophoresis and stockholders, isoelectric focusing.
Enzymes are antigenic, and since problems occurred in the late 1960s when manufacturing workers exhibited severe allergic responses after breathing enzyme dusts, procedures have now been implemented to reduce dust formation. These involve supplying enzymes as liquids wherever possible, or increasing the particle size of dry powders from 10 ?m to 200–500 ?m by either prilling (mixing the Kissland by The Essay, enzyme with polyethylene glycol and preparing small spheres by atomization) or marumerizing (mixing the enzyme with a binder and water, extruding long filaments, converting them into spheres in a marumerizer, drying them and covering them with a waxy coat). Although many industrial processes, such as cheese manufacturing, have traditionally used impure enzyme sources, often from animals or plants, the development of Is Killing much of modern industrial enzymology has gone hand in hand with the commercial exploitation of microbial enzymes. These were introduced to the West in around 1890 when the Japanese scientist Jokichi Takamine settled in the U.S.A. and set up an enzyme factory based on Japanese technology. The principal product was Takadiastase, a mixture of amylolytic and proteolytic enzymes prepared by cultivating the fungus Aspergillus oryzae on Kissland Weekend, rice or wheat bran. Takadiastase was marketed successfully in the U.S.A. as a digestive aid for the treatment of dyspepsia, which was then believed to result from the incomplete digestion of starch. Bacterial enzymes were developed in France by Is Killing Essay, August Boidin and Jean Effront, who in 1913 found that Bacillus subtilis produced a heat-stable ?-amylase when grown in a liquid medium made by extraction of malt or grain. The enzyme was primarily used within the textile industry for the removal of the starch that protects the warp in the manufacture of cotton.
In around 1930 it was found that fungal pectinases could be used in the preparation of fruit products. In subsequent years, several other hydrolases were developed and sold commercially (e.g. pectosanase, cellulase, lipase), but the technology was still fairly rudimentary. After World War Two the fermentation industry underwent rapid development as methods for the production of antibiotics were developed. These methods were soon adapted for Kissland Weekend the production of enzymes. In the 1960s, glucoamylase was introduced as a means of between the outsiders book and movie hydrolysing starch, replacing acid hydrolysis. Subsequently, in the 1960s and 1970s, proteases were incorporated into detergents and then glucose isomerase was introduced to produce sweetening agents in Kissland by The Essay, the form of high-fructose syrups.
Since the 1990s, lipases have been incorporated into washing powders, and a variety of immobilized enzyme processes have been developed (see section on enzyme immobilization), many of which utilize intracellular enzymes. Currently, enzymes are used in four distinct fields of commerce and Is Killing, technology (Table 6): • as industrial catalysts. • as therapeutic agents. • as analytic reagents. • as manipulative tools (e.g. in genetics). Of the thousands of different types of Weekend Essay enzymes, about 95% are available from suppliers in quantities ranging from ?g to kg, provided essentially for research purposes. Around 40–50 enzymes are produced on an industrial scale (i.e. ranging from multiple kilograms to tonnes per annum).
The global enzyme market is currently dominated by the hydrolases, especially the proteases, together with amylases, cellulases and lipases supplied either as liquid concentrates or as powders or granules that release the soluble enzyme on dissolution. Global production is dominated by two companies, which between them supply more than two-thirds of the global enzyme market, namely the Analysis Everyone Text, Danish company Novozymes, with a market share of 47%, and the U.S. Kissland By The Weekend Essay. company DuPont (which recently acquired Genencor), with 21%. The value of the world enzyme market has increased steadily from similarities between the outsiders book, ?110 million in 1960 to ?200 million in by The Essay, 1970, ?270 million in 1980, ?1 000 million in 1990 and over ?2 000 million in noise, 2010. Food and beverage enzymes represented the Kissland by The Weekend, largest sector of the industrial enzymes market in 2010, with a value of ?750 million, and the market for enzymes for technical applications (including diagnostic applications, research and biotechnology) accounted for a further ?700 million. Estimates of future demand are in the range of ?4 000–5 000 million between 2015 and short groom, 2016, growing at a rate of Kissland Weekend Essay 6–7% annually. The developing economies of the Asia-Pacific Region, the Middle East and Africa are now seen to be emerging as the fastest growing markets for industrial enzymes. Microbial enzymes are typically produced in batches by culturing the producing organism within a batch fermenter. Fermentation typically lasts between 30 and 150 h, with the optimum enzyme yield for the process falling somewhere between the optimum biomass yield and the point of maximal enzyme activity within the cells. Analysis Of Tina Rosenberg's Everyone Text. Relatively small fermenters with a volume of 10–100 m 3 are generally employed, allowing flexibility where a number of different products are being produced.
Many production systems are optimized by means of a fed-batch process, in which substrates are gradually fed into the reactor over the course of the by The Weekend Essay, fermentation, rather than being provided all at once at between the outsiders and movie the start of the process. True continuous culture techniques have been used in laboratory-scale studies, but have not been widely implemented on a commercial scale, although Novozymes does have a continuous process for the production of glucose isomerase, since this is a larger-volume market and the company has a very strong market share. During the production of commercially important products via enzymatic catalysis, soluble enzymes have traditionally been used in batch processes that employ some form of stirred-tank reactor (STR). In these processes, at Essay the end of the batch run the product must be separated from physiological noise, any unused substrate, and Kissland by The, also from the enzyme catalyst. Removal of the enzyme at this stage can be achieved by thermal denaturation (only if the product is thermostable) or by ammonium sulfate precipitation or ultrafiltration. Noise Definition. These processes represent a costly downstream processing stage and generally render the Kissland by The Essay, enzyme inactive, so when a new batch run is to be started a fresh batch of enzyme is required. Immobilized enzyme systems, in physiological, contrast, ‘fix’ the Kissland Weekend, enzyme so that it can be reused many times, which has a significant impact on production costs. As a very simple example, if an enzyme is His Bromance Is Killing, mixed with a solution of warm (but not too hot) agar and this is Essay, allowed to set, the enzyme will be entrapped (for the physiological noise definition, purposes of this example let us ignore the fact that the enzyme will gradually leak out of this gel). The agar can then be cut up into cubes and these can be placed in a STR, together with substrate, as shown in Figure 12. Again the reaction would be allowed to proceed (and it might actually be slower due to diffusional constraints and Kissland by The Weekend, other effects described later).
At the end of the batch run the catalyst can now be easily separated from the product by passing the reactor contents through a coarse mesh. Immediately an important downstream processing step has been carried out and, just as importantly, the active enzyme has been recovered so that it can be reused for Analysis of Tina Everyone Speaks Essay the next batch run. By The Weekend Essay. This ease of separation of enzyme from product is a major advantage of all immobilized systems over their counterparts that use free (i.e. soluble) enzyme. This physical advantage of ease of stockholders vs stakeholders reuse of immobilized biocatalysts is one of the main reasons why such systems are favoured commercially. However, immobilization may also produce biochemical changes that lead to Kissland by The Essay enhanced biocatalyst stability, which may be manifested as: • an physiological, increased rate of catalysis.
• prolonged duration of catalysis. • greater operational stability to extremes of Kissland Essay pH, temperature, etc. The particular advantage(s) conferred by immobilization will therefore differ from one system to another. Muscular System. It should be noted that often there may be no biochemical advantage at all, and the simple physical advantage of ease of separation of the biocatalyst from the Kissland by The Weekend, product may be sufficient to favour the vs stakeholders, commercial development of an immobilized process. At this point one problem that will immediately spring to mind for most students is that they have always been taught to fully mix all of the reagents of a reaction, yet the basic principle of immobilization is to by The Weekend partition the biocatalyst into a distinct phase, rather than mix it homogeneously with the substrate. Will this not cause reaction rates to be low? The answer to this question is yes, and the relationship between the activity of an immobilized system and a non-immobilized system can be expressed as the effectiveness factor (?), where: Thus an immobilized system with an effectiveness factor of 0.1 would show only 10% of the activity of a non-immobilized system with the same amount of enzyme and operating under the same conditions. At first sight this might appear to be a major problem. However, if it is possible to reuse the biocatalyst many times this is still economically viable, even with systems that have a low effectiveness factor.
In principle, therefore, for economic viability: Thus if an immobilized system has an effectiveness factor of 0.1 (i.e. Function. 10%) and we can reuse the biocatalyst 10 times, we essentially achieve the same overall catalytic activity with both the non-immobilized system and the immobilized one. However, if we are able to Essay reuse the biocatalyst 100 times we in the outsiders and movie, fact obtain 10 times more total activity from the immobilized system than from the equivalent non-immobilized system, so the immobilized system may be economically preferable. Once a biocatalyst has been immobilized it can also be put in a range of Kissland Weekend continuous-flow reactors, enabling a continuous supply of substrate to be turned into product as it passes through the reactor. The control of stockholders such continuous-flow reactors can be highly automated, leading to considerable savings in production costs. For example, a STR can be easily modified to produce a continuous-flow stirred-tank reactor (CSTR) (Figure 13a), in which the enzyme is Essay, held within the reactor by a coarse mesh, and the product continuously flows out noise of the reactor as substrate is pumped in. It is also possible to by The Essay produce a packed-bed reactor (PBR) (Figure 13b), in which the agar cubes are packed into a column and the substrate is pumped through the bed without any need for stirring. CSTRs and PBRs enable the enzyme to short speech be reused many times before it needs to be replaced. For example, in the production of high-fructose syrups, the by The Essay, immobilized glucose isomerase enzyme would typically be used continuously for between 2 and book and movie, 4 months, and only after this time (when its activity would have dropped to Kissland by The Essay 25% of the original level) would it need to be replaced. The overall operating costs of continuous-flow reactors are often significantly lower than those of equivalent batch processes.
Batch reactors need to be emptied and refilled frequently at regular intervals. Not only is this procedure expensive, but it also means that there are considerable periods of time when such reactors are not productive (so-called ‘downtime’). In addition, batch processes make uneven demands on both labour and services. They may also result in pronounced batch-to-batch variations, as the reaction conditions change with time, and they may be difficult to scale up, due to the changing power requirements for efficient mixing. Due to His Bromance Is Killing their higher overall process efficiency, continuous processes using immobilized enzymes may be undertaken in production facilities that are around 10 to 100 times smaller than those required for Weekend equivalent batch processes using soluble enzymes. Therefore the capital costs involved in setting up the facility are also considerably lower. It should be noted that although the agar entrapment method described here has provided a useful example, it is not a particularly effective form of immobilization. Vs Stakeholders. The high temperature required to prevent the agar from by The Essay, setting may lead to Is Killing thermal inactivation of the enzyme, and Kissland by The, the agar gel itself is very porous and will allow the noise, enzyme to leak out into the surrounding solution. There are in fact thousands of different techniques of immobilization, all of which are much more effective than our example.
In general these techniques can be classified as belonging to one of three categories (Figure 14): The physical adsorption of an enzyme to Kissland Essay a supporting matrix is the oldest method of immobilization. As early as 1916, J.M. Nelson and Edward G. Griffin described the adsorption of yeast invertase on to activated charcoal, and the subsequent use of this preparation for sucrose hydrolysis. Over the years a variety of similarities between book and movie adsorbents have been used, including cellulose, Sephadex, polystyrene, kaolinite, collagen, alumina, silica gel and glass.
Such immobilization procedures are extremely easy to perform, as the adsorbent and enzyme are simply stirred together for a time (typically minutes to hours). The binding forces that immobilize the catalyst on the support may involve hydrogen bonds, van der Waals forces, ionic interactions or hydrophobic interactions. Such forces are generally weak in comparison with covalent bonds—for example, a hydrogen bond has an energy content of about 20 kJ mol ?1 , compared with 200–500 kJ mol ?1 for a covalent bond. Thus, when using such methods, yields (i.e. the amount of enzyme bound per unit of adsorbent) are generally low. In addition, adsorption is generally easily reversed, and can lead to desorption of the Essay, enzyme at a critical time.
However, despite these limitations, such a method was used in the first commercial immobilized enzyme application, namely DEAE–Sephadex-immobilized l -amino acid acylase, in physiological definition, 1969. DEAE–Sephadex is an ion-exchange resin that consists of an inert dextran particle activated by the addition of numerous diethylaminoethyl groups. Particles of this material remain positively charged at pH 6–8 (see Figure 15a) and thus bind strongly to proteins, which are generally negatively charged in this pH range. If the pH is kept constant, the enzyme and support will remain ionically linked. However, when over time the enzyme loses its activity through denaturation, the pH can be adjusted to a more acidic value, the old enzyme will be desorbed, and the pH can then be readjusted back to pH 6–8 and a fresh batch of enzyme bound. Thus the support matrix may be used many times, giving the Weekend Essay, process significant economic benefits. Clearly DEAE–Sephadex immobilization is only of value for enzymes that have a neutral-to-alkaline pH optimum. Noise Definition. For enzymes that function best under acidic conditions, CM–Sephadex is more suitable.
This contains carboxymethyl groups that remain negatively charged at pH 3.5–4.5 (Figure 15b). Proteins at this pH are generally positively charged and will thus ionically bind to the support. Desorption of the enzyme will occur when the pH is adjusted to a more alkaline value. Due to the simplicity and by The Weekend, controllability of this immobilization procedure, combined with the economic benefits of reuse of the support, ion-exchange materials are now widely used as the method of choice in definition, many industrial settings. Immobilization of enzymes by covalent bonding to activated polymers is a widely used approach since, although it is often a tedious procedure, it is capable of producing an immobilized enzyme that is firmly bound to its support. Weekend Essay. The range of polymers and chemical coupling procedures that are used is enormous. The history of covalent bonding for enzyme immobilization dates back to 1949, when F. Michael and J. Ewers used the azide derivative of carboxymethylcellulose to immobilize a variety of proteins. Activated cellulose supports continue to be popular due to their inherent advantages of high hydrophilicity, ready availability, potential for derivatization, and the ease with which cellulose-based polymers can be produced either as particulate powders or as membranous films.
It is often more effective not to build the reactive group into the cellulose itself, but instead to use a chemical ‘bridge’ between the cellulose and the enzyme molecule. The requirements for such a bridging or linking molecule are that it must be small, and that once it has reacted with the support it must have a further reactive group capable of reacting with the Is Killing the Romance, enzyme. An example of such a bridging molecule is glutaraldehyde, which contains two aldehyde groups, one at either end of its (CH 2 ) 3 moiety. At neutral pH values the aldehyde groups will react with free amino groups. Thus one end of the Kissland Weekend Essay, glutaraldehyde molecule may be attached to the support, and similarities the outsiders book, the other to the enzyme. Covalently immobilized enzymes are strongly bound to their support, so when the proteins denature they are difficult to Kissland by The Weekend remove (in contrast to adsorption, as described earlier). System Function. Therefore it is usual for both the enzyme and the support to be replaced. This may result in higher operational costs compared with adsorption techniques in which the support may be reused.
The entrapment of an by The Weekend, enzyme can be achieved in a number of ways: • inclusion within the matrix of a highly cross-linked polymer. • separation from the bulk phase by a semi-permeable ‘microcapsule’ • dissolution in a distinct non-aqueous phase. An important feature of stockholders entrapment techniques is that the Kissland by The, enzyme is not in fact attached to anything. Consequently there are none of the muscular function, steric problems associated with covalent or adsorption methods (i.e. the possibility of the enzyme binding in such a way that its active site is obstructed by Essay, part of the supporting polymer matrix). The example of an enzyme retained in agar, described earlier, is a useful illustration of entrapment. A preferable alternative involves mixing the catalyst with sodium alginate gel and extruding this into Rosenberg's Essay, a solution of calcium chloride to produce solid calcium alginate particles. This technique has the advantage of not requiring the use of high temperatures. However, although it is a popular activity in by The, teaching laboratories, outside that setting it is generally unsuitable for the immobilization of purified enzymes, as these are often able to leak out of the gel. Entrapment techniques for purified enzymes are more likely to involve retaining the enzyme behind some form of ultrafiltration membrane. However, gel entrapment procedures may be useful when dealing with larger catalysts, such as whole cells.
For example, gel-immobilized living yeast cells have been used successfully in the manufacture of champagne by Moet Chandon. Immobilization: changes in enzyme properties. Earlier in this essay it was suggested that immobilization might change the properties of an noise definition, enzyme to enhance its stability. Initially it was believed that such enhanced stability resulted from the formation of bonds between the enzyme and Weekend, the supporting matrix that physically stabilize the structure of the protein. Indeed there are some published reports that describe this phenomenon.
With regard to the stabilization of proteolytic enzymes, which often exhibit more prolonged activity in Is Killing Essay, the immobilized state, this is by The, most probably explained by the fact that such proteases in free solution are prone to Analysis of Tina Rosenberg's Everyone Speaks Text Message autodigestion (i.e. enzyme molecules cleave the peptide bonds of adjacent enzyme molecules), a process that is Kissland by The, largely prevented when they are fixed to a supporting matrix. However, the effects of immobilization are more often due to the supporting matrix changing the microenvironment around the enzyme and/or introducing diffusional constraints that modify the activity of the catalyst. Similarities Between Book. Consider, for Weekend Essay example, immobilization of the enzyme by physiological noise definition, adsorption on to a polyanionic (negatively charged) support such as cellulose. If the Weekend Essay, substrate is a cation (i.e. positively charged), it will be attracted to the support and thus to the enzyme. Stockholders. In this case the enzyme might well display higher activity, as the substrate concentration in its microenvironment would be higher than that in the surrounding bulk phase. Other cations would also be attracted, and importantly these would include H + ions. By The Weekend. Thus the microenvironment would also be enriched in H + ions, so the Analysis Rosenberg's Message Essay, pH surrounding the enzyme would be lower than the by The Weekend, pH of the bulk phase. Between The Outsiders And Movie. Consequently the enzyme would also exhibit an altered pH profile compared with that of its soluble counterpart. In addition, the immobilization matrix might act as a barrier to Kissland Weekend the diffusion of substrates, products and other molecules.
For example, if a high enzyme loading was put into a gel particle and this was then immersed in substrate solution, the substrate would diffuse into groom speech, the gel and rapidly be converted into product. Enzyme molecules entrapped deeper within the gel particle might therefore be inactive simply because they had not received any substrate to by The work on (i.e. all of the substrate was converted to product in the outer layers of the particle). Although this is obviously somewhat inefficient, it does have one useful effect. When over time the enzyme within the system denatures, the loss of activity of the enzyme in the outer part of the similarities book and movie, particle means that substrate will now diffuse deeper into Kissland Weekend Essay, the particle to noise reach the previously unused core enzyme molecules. In effect this inner reserve of Weekend enzyme will offset the loss of enzyme activity through denaturation, so the system will show little or no overall loss of activity.
This explains the observation that immobilized systems often have a longer operational lifetime than their soluble equivalents. In addition, it is of interest that enzymes bound to stockholders vs stakeholders natural cell membranes (phospholipid bilayers) within living cells will also probably demonstrate these effects, and immobilized systems thus provide useful models for Kissland by The Essay the study of such membrane-bound proteins in and movie, living cells. The major industrial processes that utilize immobilized enzymes are listed in Kissland, Table 7. Sales of immobilized enzymes peaked in 1990, when they accounted for about 20% of the Romance all industrial enzyme sales, almost entirely due to Kissland by The Weekend Essay the use of glucose isomerase for the production of vs stakeholders sweetening agents. Other commercial applications utilize penicillin acylase, fumarase, ?–galactosidase and Kissland by The, amino acid acylase. The Romance Essay. Since 2000, although there has been consistent growth in enzyme markets, few new processes employing immobilized enzymes have been introduced. The following three examples highlight many of the biochemical, technological and economic considerations relating to Kissland by The Essay the use of similarities between book and movie immobilized enzymes on Weekend, a commercial and industrial scale. Undoubtedly the most significant large-scale application of immobilized enzymes involves the production of high-fructose corn syrup (HFCS). Although most of the speech, general public believe that sucrose is responsible for the ‘sweetness’ of food and drinks, there have been significant efforts to replace sucrose with alternative, and often cheaper, soluble caloric sweetening agents. HFCS is a soluble sweetener that has been used in many carbonated soft drinks since the 1980s, including brand-name colas such as Coca-Cola and Pepsi-Cola.
HFCS is produced by the enzymatic digestion of starch derived from corn (maize). Developments in HFCS production have been most prominent in countries such as the U.S.A., which have a high capacity to produce starch in the form of corn, but which do not cultivate significant amounts of by The Essay sugar cane or sugar beet, and must therefore import either the raw products (for processing) or the Speaks Text Essay, refined sugar (sucrose) itself. Simple corn syrups can be manufactured by breaking down starch derived from corn using the enzyme glucoamylase alone or in combination with ?-amylase. By The Essay. These enzymes are cheap and can be used in muscular function, a soluble form. Since starch has to be extracted from corn at high temperatures (because starch has poor solubility at low temperatures and by The Essay, forms very viscous solutions), the process utilizes enzymes from thermophilic organisms, which have very high temperature optima. Simple corn syrup is stockholders vs stakeholders, therefore composed predominantly of glucose, which unfortunately has only Kissland by The Essay, 75% of the sweetness of sucrose. However, in order to muscular system make the syrup sweeter the enzyme glucose isomerase, which catalyses the following reaction, can be employed: This enzyme (described previously in Kissland by The Essay, the section on properties and mechanisms of speech enzyme action) will produce a roughly 50:50 mixture of glucose and fructose at equilibrium, and since fructose has 150% of the sweetness of sucrose, this glucose:fructose mixture will have a similar level of sweetness to sucrose.
However, glucose isomerase is an intracellular bacterial enzyme, and would be prohibitively expensive to use in a soluble form. This makes it an ideal candidate for use in an immobilized process. The first glucose isomerase enzyme to be isolated was obtained from Kissland Weekend Essay, species of Pseudomona s in 1957, and more useful enzymes were isolated throughout the 1960s from species of Bacillus and Streptomyces. Is Killing. In 1967, the Clinton Corn Processing Company of Iowa, U.S.A. (later renamed CPC International) introduced a batch process that utilized an immobilized glucose isomerase enzyme, and by 1972 the company had developed a continuous process for the manufacture of HFCS containing 42% fructose using a glucose isomerase enzyme immobilized on a DEAE ion-exchange support. During the late 1970s, advances in Kissland by The Weekend, enzymology, process engineering and fractionation technology led to the production of syrups with a higher fructose content, and today HFCS containing 55% fructose is stockholders vs stakeholders, generally produced, and is commonly used in soft drinks, although 42% fructose syrups are still also produced for use in some processes, including the production of bakery foodstuffs. In 2010, the U.S. Kissland By The. production of HFCS was approximately 8 million metric tons, accounting for 37% of the U.S. caloric sweetener market, and similarities between the outsiders and movie, it is estimated that today about 5% of the entire corn crop in the U.S.A. is used to produce HFCS. Within the dairy industry the production of Kissland Essay 1 kg of vs stakeholders cheese requires about 10 litres of milk, and produces about 9 litres of by The Weekend whey as a waste product. Whey is a yellowish liquid containing 6% dry matter, of which nearly 80% is lactose. The enzyme lactase (?-galactosidase) may be used to break down lactose to its constituent monosaccharides, namely glucose and galactose, which are more soluble than lactose, and muscular, have potential uses as carbon sources in microbial fermentation, and can also be used as caloric sweeteners. Valio Ltd of Finland has developed arguably the by The Weekend, most successful commercial process for the treatment of whey.
Using a lactase enzyme obtained from Everyone Speaks Text Message Essay, Aspergillus , immobilized by adsorption and cross-linked on to a support resin, whey syrups are produced that have been utilized as an ingredient in drinks, ice cream and confectionery products. The Aspergillus enzyme has an acid pH optimum of 3–5, and by operating at Weekend Essay low pH the process avoids excessive microbial contamination. Physiological. Treatment plants that utilize 600-litre columns have been built in Finland, and these are used to Kissland treat 80 000 litres of whey per day. This technology has also been used to produce whey syrups in England (by Dairy Crest) and in vs stakeholders, Norway. Similar technology can also be used to remove lactose from milk. Lactose-free milk is produced for consumption by Kissland Essay, those who have lactose intolerance (a genetic condition), and also for consumption by pets such as cats, which are often unable to digest lactose easily. The first industrial processing facility to use immobilized lactase to muscular treat milk was opened in 1975, when Centrale del Latte of Milan, Italy, utilized a batch process in which yeast ( Saccharomyces ) lactase, with a neutral pH optimum of 6–8, was immobilized within hollow permeable fibres.
This process was capable of treating 10 000 litres of milk per day, and was operated at low temperature to by The Weekend Essay prevent microbial contamination. Production of semi-synthetic penicillins. High yields of natural penicillins are obtained from species of the fungus Penicillium through fermentation processes. However, over the years many microbial pathogens have become resistant to natural penicillins, and are now only treatable with semi-synthetic derivatives. These are produced through cleavage of natural penicillin, such that the G or V side chain is removed from the 6-aminopenicillanic acid (6-APA) nucleus of the molecule: Thereafter, by attachment of a chemically different side chain, a semi-synthetic penicillin product (e.g. ampicillin, amoxicillin) can be formed. In addition, the 6-APA can undergo chemical ring expansion to yield 7-aminodesacetoxycephalosporanic acid (7-ADCA), which can then be used to generate a number of important cephalosporin antibiotics (e.g. Vs Stakeholders. cephalexin, cephradine, cefadroxil). The development of immobilized penicillin G acylase dates back to Essay research conducted in 1969 by University College London and Beecham Pharmaceuticals in the U.K. Penicillin G acylases are intracellular enzymes found in E. coli and a variety of other bacteria, and the Beecham process immobilized the E. coli enzyme on a DEAE ion-exchange support.
Later systems used more permanent covalent bonding to attach the enzyme to the support. In the 1980s and 1990s, world production of penicillins was dominated by physiological, European manufacturers, which accounted for production of Kissland Essay around 30 000 tonnes of penicillin per annum, 75% of which was used for the manufacture of semi-synthetic penicillins and cephalosporins. However, over the past 10 years, due to increasing costs of labour, energy and stockholders vs stakeholders, raw materials, more bulk manufacturing has moved to the Far East, where China, Korea and India have become major producers. The market currently suffers from significant overcapacity, which has driven down the unit cost of penicillin and cephalosporin products. However, penicillins and cephalosporins still represent one of the world's major biotechnology markets, with annual sales of about ?10 000 million, accounting for 65% of the entire global antibiotics market. Enzymes have a wide variety of uses in by The Weekend, analytical procedures. Their specificity and potency allow both detection and amplification of a target analyte. ‘Wet chemistry’ enzyme-based assays for the detection and quantification of a variety of substances, including drugs, are widespread. Enzymes also play a key role in immunodiagnostics, often being used as the agent to amplify the signal—for example, in enzyme-linked immunosorbent assays (ELISAs). Within DNA-fingerprinting technology, the stockholders vs stakeholders, enzyme DNA polymerase plays a key role in the amplification of by The Weekend Essay DNA molecules in the polymerase chain reaction. Physiological Noise. However, ‘wet chemistry’ analytical methods are increasingly being replaced by the use of biosensors—that is, self-contained integrated devices which incorporate a biological recognition component (usually an immobilized enzyme) and an electrochemical detector (known as a transducer). Much of the technological development of biosensors has been motivated by the need to measure blood glucose levels.
In 2000, the by The Weekend, World Health Organization estimated that over 170 million people had diabetes, and physiological, predicted that this figure will rise to over 360 million by Kissland by The Weekend Essay, 2030. In view of this, many companies have made significant investments in RD programmes that have led to the availability of a wide variety of glucose biosensor devices. In 1962, Leland Clark Jr coined the term ‘enzyme electrode’ to describe a device in which a traditional electrode could be modified to respond to other materials by the inclusion of a nearby enzyme layer. Clark's ideas became a commercial reality in 1975 with the successful launch of the short groom speech, Yellow Springs Instruments (YSI) model 23A glucose analyser. This device incorporated glucose oxidase together with a peroxide-sensitive electrode to measure the hydrogen peroxide (H 2 O 2 ) produced during the following reaction: In this device, the by The Weekend Essay, rate of H 2 O 2 formation is noise, a measure of the rate of the reaction, which depends on the concentration of glucose in solution, thus allowing the latter to be estimated. As was discussed earlier, in enzyme-catalysed reactions the relationship between substrate concentration and reaction rate is not linear, but hyperbolic (as described by Weekend Essay, the Michaelis–Menten equation).
This is also true for the glucose oxidase within a biosensor. However, we may engineer a more linear relationship by ensuring that the enzyme is either behind or within a membrane through which the glucose must diffuse before it reacts with the enzyme. This means that the system becomes diffusionally, rather than kinetically, limited, and the response is then more linearly related to the concentration of of Tina Rosenberg's Text glucose in solution. Over the years the Kissland Weekend, YSI model 23A glucose analyser has been replaced by a range of much more advanced models. The current YSI model 2900 Series glucose analyser is shown in Figure 16. This instrument has a 96-sample rack that enables batches of samples to function be run, with the Weekend, analysis of each sample taking less than a minute.
The instrument can measure the glucose content of whole blood, plasma or serum, and requires only 10 ?l of sample per speech, analysis. The membrane-bound glucose oxidase typically only needs to be replaced every 3 weeks, thereby reducing the cost of Essay analysis. These systems also offer advanced data-handling and data-storage facilities. In addition, these instruments can be modified to analyse a wide variety of noise other substances of biological interest, simply by Weekend Essay, incorporating other oxidase enzymes into the membrane (Table 8). To enable diabetic patients to take their own blood glucose measurements, small hand-held biosensors have also been developed, which are in fact technologically more advanced because the enzyme and transducer are more intimately linked on the sensor surface.
The first device of this type was launched in 1986 by Rosenberg's Message, Medisense, and was based on technology developed in the U.K. at Cranfield and Oxford Universities. The ExacTech blood glucose meter was the size and shape of a pen, and used disposable electrode strips. Kissland By The Weekend. This device was followed by a credit card-style meter in 1989. Such devices again rely on glucose oxidase as the biological component, but do not measure the reaction rate via the stockholders vs stakeholders, production (and detection) of H 2 O 2 . Kissland Essay. Instead they rely on direct measurement of the rate of electron flow from of Tina Everyone Speaks Text Message, glucose to the electrode surface. The reactions that occur within this device may be summarized as follows: and at the electrode surface: where GO x -FAD represents the FAD redox centre of glucose oxidase in its oxidized form, and GO x -FADH 2 represents the reduced form.
Basically electrons are removed from the glucose molecules and passed via the enzyme to the ferrocene mediator, which then donates them to by The Weekend Essay the working electrode surface, resulting in the generation of an physiological noise definition, electrical current that is Kissland Weekend, directly proportional to between and movie the rate of oxidation of glucose, and thus proportional to the glucose concentration in the sample. Medisense, whose only product was its blood glucose meter, was bought by Abbott Diagnostics in 1996, and Abbott-branded devices continued to use and develop this technology for some time. In 1999, Therasense marketed a glucose meter that represented the next generation of sensing technology, and integrated the enzyme even more closely with the Weekend Essay, electrode. Originally developed by Adam Heller at book the University of Texas in the 1990s, wired-enzyme electrodes do not rely on a soluble mediator such as the ferrocene used in Essay, the Medisense devices. Instead the enzyme is immobilized in an osmium-based polyvinyl imidazole hydrogel in which the electrons are passed from enzyme to Analysis Rosenberg's Speaks Essay electrode by a series of Weekend Essay fixed electroactive osmium centres that shuttle the electrons onward in a process called ‘electron hopping.’ In 2004, Abbott Diagnostics purchased Therasense, and instruments such as the FreeStyle Freedom Lite meter range produced by Abbott Diabetes Care (Figure 17) now incorporate this wired-enzyme technology. Devices of this type are highly amenable to between the outsiders book miniaturization. Continuous measuring devices are becoming increasingly available, and may well revolutionize the control of Kissland Weekend Essay certain disease conditions. For example, with regard to diabetes, devices such as the FreeStyle Navigator range from Abbott Diabetes Care use the same wired-enzyme technology as that described earlier, but now incorporate this into a tiny filament about the diameter of the outsiders book a thin hypodermic needle. This is inserted approximately 5 mm under the skin to measure the glucose level in Essay, the interstitial fluid that flows between the cells. Muscular System Function. The unit is by The Essay, designed to remain in situ for up to 5 days, during which time it can measure the glucose concentration every minute.
A wireless transmitter sends the glucose readings to a separate receiver anywhere within a 30-metre range, and this can then issue an early warning alarm to alert the muscular system, user to a falling or rising glucose level in time for them to Essay take appropriate action and avoid a hypoglycaemic or hyperglycaemic episode. In addition, experimental units have already been developed that link continuous glucose biosensor measurement systems with pumps capable of gradually dispensing insulin such that the diabetic condition is automatically and reliably controlled, thereby avoiding the traditional peaks and troughs in glucose levels that occur with conventional glucose measurement and the intermittent administration of insulin. Therefore, looking to the future, we may confidently expect to see the development of biosensor systems that can continuously monitor a range of physiologically important analytes and automatically dispense the required medication to alleviate the symptoms of stockholders vs stakeholders a number of long-term chronic human illnesses. For the sake of conciseness, this guide has been limited to some of the basic principles of enzymology, together with an overview of the biotechnological applications of enzymes. Essay. It is important to understand the relationship between proteins and the nucleic acids (DNA and RNA) that provide the blueprint for the assembly of proteins within the cell. Genetic engineering is thus predominantly concerned with modifying the proteins that a cell contains, and genetic defects (in medicine) generally relate to groom speech the abnormalities that occur in Kissland Essay, the proteins within cells. Much of the between, molecular age of biochemistry is therefore very much focused on Kissland, the study of the cell, its enzymes and other proteins, and vs stakeholders, their functions. Abbreviations7-ADCA, 7-aminodesacetoxycephalosporanic acid; 6-APA, 6-aminopenicillanic acid; ATCase, aspartate transcarbamoylase; CSTR, continuous-flow stirred-tank reactor; CTP, cytidine triphosphate; DFP, diisopropyl fluorophosphate; EC, Enzyme Commission; ELISA, enzyme-linked immunosorbent assay; GUT, glucose-1-phosphate uridylyltransferase; HFCS, high-fructose corn syrup; PBR, packed-bed reactor; PFK, phosphofructokinase; STR, stirred-tank reactor. This article is a reviewed, revised and updated version of the following ‘Biochemistry Across the School Curriculum’ (BASC) booklet: Teal A.R. Wymer P.E.O., 1995: Enzymes and their Role in Technology.
For further information and to provide feedback on this or any other Biochemical Society education resource, please contact education biochemistry.org . For further information on other Biochemical Society publications, please visit www.biochemistry.org/publications. This is an Essay, open access article published by Portland Press Limited and distributed under the Creative Commons Attribution Licence 3.0.