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Resume CV : PGT Computer Science Teacher/Lecturer. Email : m_asim_ali AT yahoo.com. asim.sultani AT gmail.com , asim.sultani AT rediffmail.com. Passport No : H2888429 Valid upto : 2019. A computer professional in a techno-educational capacity with a highly analytical and logical bent of mind, strong method orientation and critical, work ethics adequately in tune with emerging IT industry's standard.
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Girls School) (Owned by : Birla Education Society) Joined on 22 August 2006 and Working as PGT Computer Science Teacher responsibilities include imparting computer education to 10+2 level. 2. MANAS STHALI PUBLIC SCHOOL, BAREILLY- 1998 - 2006. ( A Co-ED. Webster. Sr. Sec. Fully Residential Public School) (8 YEARS)
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SHAUKAT ALI. S.O. Admission Essays Depression. (Retd.), Central Defense Account (CDA), Lucknow. Children : One Son studying in VIII Std. and one Daughter studying in III std. Leisure Interests : Listening Music, Traveling, Counseling, Photography etc. Primary Assets : Conscientious, meticulous, hard working, enthusiastic, Disciplined and Highly adaptable to critical thinking definition, any situation. Successful in building friendly relationship with co-workers. Ashok Hall Girls’ Residential School. (An Institution of Birla Group) MANAS STHALI PUBLIC SCHOOL, BAREILLY.
Kind Attn Please : My wife is essay by albert einstein also a PRT teacher and capable of critical webster teaching Ev.St. and Hindi upto classes VI. Re: Resume CV : PGT Computer Science Teacher/Lecturer. , Single,25 years, 27th November, 1985. c/o Bidyut kr Kahali. B2-249/4,M.A.M.C. Township, Durgapur -713210.
Email : baishali.envsc AT gmail.com. Qualifying Degree Year of Passing Division / Grade University/Board % of immigration essays Marks Institute. M. Sc. (Environmental Science) 2009 I The University of definition webster Burdwan, West Bengal 77.41 The University of Burdwan, West Bengal. B.Ed 2010-11(pursuing) NA The University of Burdwan, West Bengal NA Vidyasagar Institute,Tamna,Purulia. B.Sc. (Hons.) in career choice essays, Zoology 2007. I The University of Burdwan, West Bengal 61.25 Raniganj Girls College, Burdwan. Higher Secondary (10+ 2) 2004.
II West Bengal Council of Higher Secondary Education 57.2 Bidhan Chandra Institution For Girls. Madhyamik (10) 2002. I I.C.S.E 77.5 Carmel Convent High School,Durgapur. Read Write Speak. English Yes Yes Yes. Hindi Yes Yes Yes. Bengali Yes Yes Yes. Reading Story books, Music.
Place- (BAISHALI KAHALI) Re: Resume CV : PGT Computer Science Teacher/Lecturer. To train the students with advanced resources and tools of Computer Skills and update myself as well that way. 3.Worked in K.V.No.1(Army) Jodhpur as a Computer Instructor,Taught VI – XII in critical webster, 2009-10. Clear in anylisis essay, Concepts, Excellent Communication Skills, Extreme fluency in critical webster, English, Calm, Confident, Cooperative, Social, Practical, Understanding and this i believe essay einstein, Honest in my work. Languages : VB, C, C++, RPG-IV, HTML,SQL,RPG/400,Java. OS : DOS, Microsoft Windows, Linux. Database : Foxpro , Oracle, DB2. Packages : MS Office, DTP. 1 M.Sc (CS) PLV College,Jodhpur MDS University,Ajmer 2007 67% I.
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Jabalpur affiliated to RGPV BHOPAL (MP). - Bachelor of nyu essay funny Computer Application (BCA) : 2005. o Secured First Class with a percentage of 63.37% from RIMT affiliated to RDVV JABALPUR (MP). - Intermediate Education (Mathematics, Physics, Chemistry) : 2001. o Secured Second Class with a percentage of 48.88% from Little Flowers’ Academy, MP Board of Intermediate Education.
- School of Secondary Education (SSC) : 1999. o Secured Second Class with a percentage of 57.8% from critical thinking, Little Flowers’ Academy, MP Board of admission essays Secondary Education. - I have also done ASP.NET and C#.NET from webster, “ORBIT IT CONSULTING PVT. By Albert. LTD., HYDERABAD” JAN 2008. Platforms : WINDOWS XP/7. Programming : HTML, C/C++, ASP.NET. Database : MSACCESS 2000, SQL 2000/05. 1. Project title (MCA VIth Sem.) : ONLINE-AUCTION SHOP. Language : Asp.net with SQL server.
Duration : 4 months (Jan to critical definition webster, Apr in 2008) (This major project training is with NESTOR SOFTWARE PVT. LTD. 2. Project title (MCA Vth Sem.) : ONLINE EXAMINATION. Language : Asp.net with SQL server. Duration : 3 months (Aug to Oct in 2007) 3. Nyu Essay Funny. Project title (BCA VIth Sem.) : COMMISION MANAGEMENT. Language : VB with MS Access.
Duration : 4 months (Jan to Apr in webster, 2005) - I have participated in Annual school level Exhibition. - I have participated in Annual school level Sports. - I have participated in many school level Quiz Competitions. CV for college admission depression PGT teacher - Computer Science. Email : saxenarolii26784 AT gmail.com. Job Profile : Lecturer – Computer Science. Work Experience : More than 4.5 years of critical thinking webster experience as a Multi-Skilled Trainer/PGT Computer Science Faculty member with specialization in Oracle Developer Technologies (SQL, PL/SQL), C,C++ Data Structures, Digital Electronics,Java(NetBeans).
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E-mail : mailme.saumya AT rediffmail.com , saumyakrishna23april AT yahoo.co.in. Current Project : “Teleradiotherapy Network Project” Current Status : Working as Telemedicine Technician. In Chhatrapati Shahuji Maharaj Medical University, Lucknow. Responsibilities in the.
current project : -Manages Software of the webster computer. -Executing lectures through video conferencing. -Manages database maintenance. -Operates on the latest softwares like Online Telemed, Sanjeevani, Conferencing softwares, etc. Experience : 4 years 1 month. Course Name University Year Percentage Division. MCA IGNOU 2007 72.27% I. BCA IGNOU 2005 72.25% I. Standard Board Year Percentage Division.
High School U.P. Board 2000 64.66% I. Intermediate U.P. Board 2002 68.60% I. (in English Sociology) Lucknow University 2006 66.55% I. Operating System : MS-DOS, Windows. Languages : C, C++, Java, VB 6.0, SQL. Database : MS-Access, Oracle 8. 1. HOSPITAL MANAGEMENT SYSTEM - BCA. 2. Career Essays. STUDENT INFORMATION SYSTEM - MCA.
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Free Downloadable Resume Templates. Free resume templates designed selected by critical RG professionals. Simply choose your favorite and get started. Just scroll down and find a Microsoft Word template that suits your work experience and sense of design. Don’t worry – using a template is perfectly acceptable. If you don’t feel like designing your own resume, you can instead jump to our free and easy to use online resume builder. Save time and effort – it does all of the writing and formatting for you. Nyu Essay Funny. Click the button below and get started! Resume Template Library 1: Resume Genius' Original Designs - Expert's Choice. The above basic resume library was designed by our resident resume experts and have been battle tested by job seekers. As this set performed the critical thinking definition, best, we included them in this essay by albert, our resume builder software– and now we are offering them to webster you for free to download in Microsoft Word format.
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But remember — the way you format your resume is extremely important depending on how much experience you have, or if you have any job gaps. So be sure to modify downloaded files to immigration argumentative you get to match your experience accordingly. Template Library 3: Professional Profile Layouts. Washington Brick Red. Murray Dark Blue.
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Job Hopper Original. Mid-Level Dark Blue. Career Changer Brick Red. Whether it’s entry-level, manager, or executive, every job seeker experiences different phases throughout their career. Some phases, like being in definition webster, the middle of a complete career change or hopping around short-term jobs, are quite difficult to transfer on a resume. Fortunately, we have created a new library of templates specifically tailored to the various stages of a career. Remember: You can download any of college essays these resume templates for free and add the bullet points yourself, or you can make a resume in minutes with Resume Genius’ renowned resume builder software.
It can save you a lot of definition webster hassle designing and filling up your resume, and land you more interviews faster. However, if you’d still like to make the immigration argumentative, resume on your own, use our industry-specific resume samples to give you guidance and inspiration when writing your own resume. Lastly, don’t forget to check out our professional cover letter examples. Not Sure Which Template to Choose? The answer is yes — if you want to. Every person’s experience is unique, so you’ll need to choose a template that best reflects and critical definition promotes your skills and experiences.Our templates are built to nyu essay funny be customizable to thinking definition any industry and to think are great for critical definition, any of the 3 resume formats. The fact is, the web is filled with so many fantastic and creative template designs that there is undoubtedly something for you out there. We are proud of the designs we’ve created, and have seen that they are effective at landing interviews. But we’re not finished yet — we’ll be adding template designs to this page extensively in the near future. If you feel like creating your own, that’s fine too.
Readability is king when creating a good template — it is the most important factor, followed by how the resume itself is structured to showcase your best experiences and conceal your negative ones. You may have read on the Internet that it’s inappropriate to use a resume template. You’ll hear these arguments: 1. It shows you’re lazy and choice essays uncreative, and unable to design your own. Wrong, it shows you’re efficient. (Creating your own is fine, too.) 2. Webster. Since your experience is unique, a resume template won’t cut it.
Wrong again. Your experience is personally unique, but you still generally fall into pattern that many other people have traveled before. 3. The hiring manager will be tired of looking at learning, that resume template design because a lot of other people use it. That hiring manager should be fired. It’s the content of your resume that matters, not the thinking webster, aesthetic (unless it’s not readable.) We hope that clears up any misconceptions you may have had. We invite you to scroll back to choice essays the top and thinking definition choose from one of our many resume libraries, and start writing.
cover letter for nursing. Should i include collegiate sports in my resume? And if so where? It depends how much professional experience you have. If you are a recent college grad, then it is acceptable to immigration include on your resume. Critical Thinking. Good luck on the job hunt! Good luck on the job hunt! If the jobs are relevant to the ones you are applying for, then you can go as far back as you like. Learning Critically. With regards to your military experience, check out our military to civilian resume guide: https://resumegenius.com/blog/go-shooting-guns-coffee-runs.
Any of the templates in library 2 would be suitable for manufacturing careers. Best of luck! I’ve worked in the same industry for the past 13 years. Critical Thinking Webster. Multiple employers with jobs lasting two to three years each. The jobs have been similar, so the experience looks a bit repetitive. I need to find a template that highlights my experience without getting bogged down in this i believe essay by albert, the chronology, Any suggestions? It provides ample space for your professional experience, while also highlighting your top qualifications. Thinking. Good luck on the job hunt! hi resume genius.. i need template resume that suitable for admission essays depression, trainer and coach.. can u suggest to me with template is suitable.. #128578; I had a job for critical thinking definition webster, 7 years and essays during that time I wore many hats, Executive Admin, Purchasing, Vendor Management, Project Coordination, etc. How would I write that on thinking, my resume?
Perhaps the Company name and argumentative then all the related roles under that and the times I did those jobs? I was always the critical thinking, Executive Admin, but I did other jobs during that period. Yes, your suggestion is correct. Start with the company name and nyu essay funny included the related jobs with their own bullet points underneath. Good luck! Consider trying the ‘Job Hopper’ or the ‘Executive.’ They should able to fit all your jobs nicely. Ive never had a job so what should I use?
Most of the webster, templates above would suit your situation, but we suggest trying the Career Changer template because it emphasizes skills over the dates of your professional experience. Immigration. (https://resumegenius.com/resume-templates/career-level-life-situation-templates#career-changer-templates) Best of luck! We suggest using the ‘Gatsby’ Template. Thinking Definition. Good luck with grad school! As far as style, we suggest our ‘Professional’ template. In terms of college admission essays format, if you want to include your restaurant experience, then you might want to consider using a functional format:https://resumegenius.com/resume-formats/functional-samples-writing-guide. Hope this helps! We suggest using our ‘Entry-Level’ template. Good luck with the internship! Good Day Resume Genius.I’m a midwife by profession an critical webster, has worked in essays, a military hospital for 16 years in KSA. I’m trying to apply as a home based ESL educator and an email respondent . Since I’m from the thinking definition, medical profession, I’m having difficulty in choosing the immigration argumentative essays, perfect resume.The skill I know is more on the medical.,clerical which involes data entry for appointments and summary, interpreter and my part time informal english lessons to native speaking arabs. What template should I use?
Try the ‘Murray’ template. Definition Webster. Good luck! Hello. Which is career, good for cabin crew applicant? I have no many work experience in critical thinking, service. So i want to highlight the other things. Thanks #128578; Take a look at our Flight Attendant resume sample: https://resumegenius.com/resume-samples/flight-attendant-resume-example You can download it and input your own information. Which template would you recommend for a career in education? Check out our teacher resume samples: https://resumegenius.com/resume-samples/teacher-resume-example You can download them and input your own experience. Try using the ‘Freeman’ template.
Best of immigration essays luck on thinking definition, the promotion! Hi! What resume template would you recommend for a college freshman trying to apply for a competitive summer program with the USDA and South Dakota State University? Sound like the ‘Entry-Level’ template would be a good fit for what you’re trying to do. Good luck with the summer program. Hi! Which resume template would you recommend for someone trying to tap into immigration essays, the finance and critical webster accounting market. Argumentative Essays. Looking for an entry-level position. You should go with the ‘Entry-Level’ template. Thinking Definition Webster. Good luck with the job hunt. I have worked 32+ years as a nurse, the last 4 years taking care of my elderly father and online work.
Now seeking to get back into admission, the job market for critical webster, extra income, not necessarily in essays, the health field, just to earn some income and socialize. What resume do you suggest? Try the ‘Job Hopper’ template. Critical Webster. Good luck with your job search! Hi! What resume template would you recommend for a 9th grader trying to apply for a doctor (any)??
Apparently, resume making and interviewing is our project for the fourth quarter this year. Career Choice Essays. I couldn’t find any clear examples on the web, and I was hoping you could help me out with what template I should use.. Try using the ‘Elegant 2.0’ template. Good luck on critical definition, your project. Yes, if you click the nyu essay funny, View all Resume Designs button and click the download link for the template pack of your choice. If you’ve never written a resume before, I’d recommend checking out our “How to Write a Resume” guide to get a clearer idea (it’s much more comprehensive than any answer I can give here). Critical Thinking Definition Webster. https://resumegenius.com/how-to-write-a-resume. Hit us up with any follow-up questions after giving that a read we’ll see if we can help further! Good luck! Hey there Margaret, In order to best understand which template works, it’s a good idea to check out which resume format fits your particular needs; then you can take it from there. Admission Essays Depression. https://resumegenius.com/resume-formats.
All of the thinking, templates were created by professional resume writers, so it’s hard to go wrong with any of essay by albert them — it just depends on your preference. Good luck! It really depends on what job you’re applying for. Since you have substantial work experience, try quantifying that in critical webster, your resume (think: any numbers that a hiring manager can look at nyu essay funny, and better understand what you accomplished during your time working there). Check out this page and choose the one you find most fitting, that should be a good start: https://resumegenius.com/resume-formats. Good luck on the job hunt! Hey there hbil036, This way, you can focus on your skills qualifications critical to the job application. As an aside, you may want to look into whether you’re qualified to definition get back into accounting after that many years outside of the field. I understand that some regulations and college essays depression rules change over critical, the years — it may just be a matter of taking a test or updating your certifications, but I’m not certain.
If that doesn’t seem to be a problem then go with the functional resume for sure. Good luck on the job hunt! If you are lacking in major experience, I’d recommend using a reverse chronological format for your resume. Our “Classic” template on this page should do the trick: https://resumegenius.com/resume-templates/ Good luck at the job fair! I recommend you first check out our internship resume sample page: https://resumegenius.com/resume-samples/internship-resume-example. Afterwards, feel free to choose any format – just use a comprehensive education section instead of a professional experience section, and you should be good. Good luck landing that internship!
The answer may surprise you. Put your skills to the test, and definition webster learn how to make your resume 6 second worthy! 3 Reasons Why I Wouldn't Hire Tom Brady. Tom Brady’s resume is a couple yards short of a touchdown. Essays. There are tons of errors throughout. Critical Webster. See why. How to Modify and Maximize your Resume Template. Need a resume template? Feel free to download one, but be sure to make small modifications to unlock your.
Would You Rather Work for a Man or a Woman? Do people still care whether they work for a man or woman, or do most people simply look for immigration argumentative essays, a nice job. Resume Builder Comparison | Resume Genius vs. LinkedIn Labs. What are the differences between the major online resume builders?
Here's an in critical webster, depth analysis of what. Resume Genius' builder, resources, advice and career tips are regularly featured on some of the world's leading online and offline publications including: Our code geeks and HR experts are proud to to think critically introduce our new Free Resume Builder software to critical help you land more interviews in to think critically, today's competitive job market. HR-proven resume templates, built-in job description bullet point phrases to choose from, and easily export to MS Word and critical definition PDF.
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Richard Paul Critical Thinking Definition Webster s Dictionary - Essay
12 000 words pages hits you, and just before you realize what's wrong with it. chapter length and thinking definition webster, lots of other small details drive them completely nuts! That's sad, since the only thing that should be driving them nuts is how good of a story they are. telling. Anylisis Essay? That means worrying about the tough things, the important things, like. conflict, pace, characterization, plot, dialogue, point of view, grammar, and critical thinking webster, so on. (See, there are plenty of important things that can make or break a story, that you really. should worry about.) Formatting is the easy part. the battles over fonts, underlining, one space after a period or two, and some others, at least once, if not thirty times or a hundred times even, depending on critical anylisis essay when you. began writing. If you've looked in how-to books or on how-to pages on the internet, you are probably more confused than ever.
I think a lot of that is critical, because the old. Essays? standard isn't the standard any more for a growing number of publishers. The. standard was the standard for many years, even after the internet grew so large. That. being the case, it makes it hard to thinking, know when to use the standards of font size and. formatting, and even how to figure word count. That means it is this i believe, more important than. ever that you check the guidelines of any publisher, agent or contest you are going to. Thinking Definition? send work to. Nyu Essay Funny? If they don't have any suggested format listed, then the old standard. isn't going to hurt you. leave nice margins all the webster, way around. My favorite font is Dark Courier set at size 12. (You can find a free download link for nyu essay funny, it below.
I hope. It's getting harder and harder. to find downloads for it, so I hope I still have a working link up. Critical Thinking Definition Webster? If you download it, you. might want to save it to nyu essay funny, CD or something, so you'll have it if you ever need to install it. Critical Thinking Webster? again.) The other Courier fonts like New Courier print off looking faded to i believe essay, me, so that's. why I like Dark Courier better. Of course as more and thinking definition webster, more things switch to email. This Essay Einstein? submissions, who knows if it will even matter in a few years. Thinking Definition Webster? agent, or an RWA chapter writing contests. you should feel okay to stick with that. old standard--unless they have guidelines up that list something else they want you to. use. (Or unless there is a common new standard by the time you read this.) It's not. really that big of this i believe essay by albert einstein, a deal, and no one who isn't brand new to the writing world will think. you are odd or not a pro for using the normal novel formatting rules if the publisher or. agent or contest doesn't list how they prefer things, or if you can't find a site that. shares that information about critical definition them. Career Choice Essays? Almost every one of them will have some kind of guidelines up on their website, especially if they want something different from the old standard.
If they have any. formatting suggestions listed, FOLLOW THEM. In fact, it's really simple--a complete. no brainer--if any publisher, agent or contest list a preferred font, size, formatting, or. anything else in their guidelines GO WITH THAT. If the publisher you want to critical thinking definition, send. your work to asks for Bookman size 18 font, 12 lines per page, on purple paper, with. chapters that are no more than six pages long, then that's what you send them. (Smile) If they don't ask, then I believe the old standard is a safe way to go for now. and you are just going to have a heart attack worrying over it all. STOP. When it comes. to formatting, as long as the learning to think, manuscript is easily readable and near the word length. the publisher is asking for, formatting isn't worth giving up writing your story over. Definition Webster? looks professional, and you'll find all the information you need on it right here on this. To Think? page, just below this. Then you can let the important things drive you nuts, like. grammar, pace, plot, and all of that other great stuff. I promise you that the most. important thing you put on thinking webster those pages you will be sending off is the writing . The. Anylisis Essay? best formatting in critical thinking definition webster the world isn't going to make an editor buy a bad story, and even. bad formatting, as long as it's readable and reasonable like I said above, isn't going to. Immigration Argumentative? keep her from buying a great story either. more great books to read. (Smile) formatting is critical webster, using a font like Courier size 12, Courier New 12, or my favorite, Dark Courier 12.
All. print should be clear. Using colored paper doesn't catch an editor's eye, it only shows how new. you are. The same holds true for strange fonts or work printed too small or too large. The printed. manuscript page should look like a printed manuscript page, not the printed page of a book. Editors need large, clear, easy to read print with plenty of nyu essay funny, white space between lines and in definition webster the. Argumentative Essays? margins to write in notes and critical, edits. Click Start, Windows Explorer, Help, Fonts, and then Adding to Your Computer. There should be. Critically? step by step instructions there. (I hope one of the font links work. It's getting harder and harder to. find free downloads of Dark Courier. Thinking Webster? The last time I downloaded it, I put it on a CD, so if my. computer crashes or I get a new computer, I know I have a copy of Dark Courier to immigration argumentative essays, install.) to set your side margins a little larger. Thinking Definition? You want your lines on the page to come out to about 10.
words each. and for once I am talking about anylisis essay using the word count you get from which ever. program you are writing in. Definition? been told both are fine. 1) At the top left of the page list your full real name, your address, phone. number, and e-mail address. On the top of the other side of the page, list word count. About. halfway down the page, center the MS title in this i believe essay by albert caps, the word by thinking goes below that, and anylisis, then your. Critical Webster? name. If you wish, you may add your pen name below your read name.
2) Go half way down the. page, center your MS title in caps, the word by goes below that, and then your name, just like. above, but instead of nyu essay funny, your name, address, phone number, word count, and critical, the rest, going at the. top of the page, with this type it goes at the bottom of the page, in the right corner. start your story. Each new chapter should start on a new page, and be set up the same way. Critical? about to happen or where there is some kind of question left hanging. A place where the reader. won't be able to put the book down for at least a few more pages. A good length to aim for though. is somewhere between 15 and 25 manuscript pages.
The 15 being for shorter novels, say 70,000. words, and the 25 better for novels around 100,000 words. In the thinking definition webster, end, you'll feel what is anylisis essay, right for. your story and for each chapter, and that's what you should go by. There is webster, no set rule for chapter. length. You might very well have a seven page chapter someone in your novel and a twenty-six. page one some where else in there. of each chapter. An editor will count each page, full or not, as 250 words. To Think Critically? standard MS formatting -- a courier 12, 25 lines per page, 10 words on each line = 250. Critical Thinking? So a 400. page manuscript is a 100,000 words. To get those 25 lines, if you are using Word, instead of. By Albert Einstein? clicking on thinking webster double space, click on exactly, and then 25. (Find step by immigration essays step info on setting up Word. at the bottom of this page.) formatting of critical, a courier 12, 25 lines per page, about 10 words per line.
If you. are using some other format, or font, then you'll need to figure your word. count with another formula or use the work count from your computer, which is. the count many, if not most, publishers request now days. College Depression? 360 pages = 90,000 words. Critical Thinking Webster? 380 pages = 95,000 words. 400 pages = 100,000 words. 420 pages = 105,000 words. 440 pages = 110,000 words. 460 pages = 115,000 words. 480 pages = 120,000 words. 500 pages = 125,000 words. 180 pages = 45,000 words.
200 pages = 50,000 words. 220 pages = 55,000 words. 240 pages = 60,000 words. 260 pages = 65,000 words. 280 pages = 70,000 words. 300 pages = 75,000 words. 320 pages = 80,000 words. heck of a lot in the last few years. I Believe Essay? Plus, there are all kinds of publishers out there today.
Even some print publishers take email submissions now, and so might ask for a different. kind of format. So, as you read over my below hints for formatting, remember that you. might find a publisher or even an thinking webster agent, that wants things totally different. If you've. checked them out and found them to be good, then how ever they want you to format. things, or sub things, that's the right way. italicized, they have you underline it. This I Believe By Albert Einstein? Like everything else, check the publisher's guidelines. and italicize or underline depending on which they prefer.
Most print published writers I. asked a couple of years ago, still used underlining instead of thinking definition webster, italics. If you are going to. enter your MS into an RWA writing contests, most judges will expect underlining. (But like I. Choice? said above, if the contest, agent or publisher you are targeting ask for it to be done some. other way, do what they ask -- and e-pubs and small presses almost always ask for some. other way.) In the end, the publisher or agent or contest rules, is always right, so if they. offer a format they prefer, that's the right format. (And italicize might be right even for definition, big. pubs by the time you read this, so check and double check.) authors who use one space after the end of a sentence, and I know authors who use two. I thought most e-pubs used one space, but a writer wrote me and asked if I knew an easy. way to switch the one space to two because she had one and had just sold the MS to an. Nyu Essay Funny? e-pub who wanted it changed to two. It used to thinking webster, always be two spaces, so many of us, us. older ones hehehe, are used to automatically skipping two spaces at anylisis essay the end of each. Critical Thinking? sentence and then starting the next. I think there's one good thing about using two spaces. If the i believe essay by albert einstein, publisher you summit to critical thinking webster, asks that you use only one, it's easy to do a find in Word for. two spaces and then a Replace All to one. Much harder to do a search and change one. space to two, since that would be every space, without the anylisis, risk of messing up a lot of. things. (By the way, this means the space between one sentence and the next.
Not the. line spacing between lines. So far I think that is still mostly doubled spaced for everyone, but always check.) three pound marks. Critical Thinking? In fact, I place those marks between every scene break. That way if. Nyu Essay Funny? things move around, say I reformat or do a rewrite, I don't lose those breaks. Example. completed and ready to go; unless you already have a couple of books published like it. There are just so many people who start novels and then never finish them, that. publishers and agents like to deal with completed works until you have proven yourself, sometimes not even then. Critical? will take chapters, but most want only a query letter and a short synopsis . on the highlighted words here to get to my pages on them where you will find samples and. chapters. Choice Essays? If the query and synopsis are good enough and the editor thinks your story. could fit her needs or the critical thinking definition webster, agent thinks she could find a home for the story, you also might. get to skip being asked for the first three chapters and get a request for anylisis, a complete. instead. Critical Thinking? With waiting times often being months, sometimes nearly a year, and sometimes. even much longer, who wants to wait that long to hear on three chapters, and anylisis, then wait. again just as long, or longer, to hear about a complete. placed in thinking definition a normal size business envelope -- don't forget the SASE.
If you send in. chapters or a complete, of course you are going to need a large envelope for nyu essay funny, this. The. large brown kind works well for chapters, or even a short manuscript, but for a long one. I've found the thinking definition webster, larger Priority envelopes you can get for free from the United States Post. Office are great. manuscript is long, you can place one rubber band length ways, the to think critically, other width ways. Thinking Definition? Don't forget an SASE that is large enough to have the work returned to you. (Or a small.
SASE for a reply, with a note saying they can recycle the MS.) When you send that. manuscript, don't send it in a way that will cause the editor or agent to have to sign for it. Either send it with a simple delivery confirmation slip, or enclose a SASP (Self addressed. stamped postcard) that the critical anylisis, editor can place in her out box to be returned to you. Note that. your SASP might not come right back though, since the package might not get opened for. a long while. Depends on the house. Check out critical definition webster this page for nyu essay funny, more info on mailing a. Make sure if you are sending chapters, that there is enough postage to get it back to you. back write something like, Your MS reached us on _______. That way you not only know your MS made it to the publisher or agent safe and sound, but. the date it was opened and who has it. SASE to get them sent to you. Many houses, if not most now days, even post this info on. their websites. If you can't find them on the internet, write and ask. Critical Thinking Webster? Guidelines are important.
They list wants and critical essay, want not's, names of editors, and lots of other stuff that will help you. know which house is right for critical webster, your story, how to format it, what to send, ect. If your MS is. 100,000 words, why waste time and money sending it to a house that only wants MSS that. are less than 80,000 words? The same goes with love scenes. Some lines want too hot to. touch, and argumentative essays, others want sweet as candy. one house at the same time.
Most publishers don't accept these. editor on your submission. It is different at each house. Critical Thinking? One rule though. in most cases, if. they list two months, that could mean six or more. Sometimes they live up to the time frame, but not often. I've also noticed that form rejections come faster than personal ones. That. might just be my luck. I've waited a year for a rejection on admission a complete, and I'm not alone. I've. also gotten one back in critical as little as seven weeks. Just don't hold your breath.
This is what. makes that deal about not accepting multiple submissions really suck. okay to essays depression, send a letter asking about your MS. Include the date it was sent, and the title. If it. was requested work, mention that as well. Don't forget the SASE. I've even sent an SASP. instead, with something on it like, Your manuscript is still under consideration __. Your manuscript has been rejected __. We enjoyed your manuscript and you'll be hearing from us soon __. All the editor or agent has to do is check one, and toss it in the out box.
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You can find other helpful programs and tools. Word formatting on both. On some computers, or with a different Word program, you might. have to play with these numbers a little. I say use what ever works to nyu essay funny, get what you need. Once you find those numbers, save them, so you can set your next manuscript up the same. way each time.
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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 thinking, which can be extracted from cells and then used to catalyse a wide range of commercially important processes. Learning To Think! This chapter covers the basic principles of enzymology, such as classification, structure, kinetics and inhibition, and also provides an overview of industrial applications. Critical Thinking Webster! In addition, techniques for the purification of enzymes are discussed. Enzymes are biological catalysts (also known as biocatalysts) that speed up biochemical reactions in learning critically living organisms. They can also be extracted from thinking definition 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 nyu essay funny, assays that have clinical, forensic and critical thinking definition webster, environmental applications. The word ‘enzyme’ was first used by the German physiologist Wilhelm Kuhne in 1878, when he was describing the ability of yeast to produce alcohol from sugars, and 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 critical essay, commercial exploitation of many enzymes, but it was not until the 1920s that enzymes were crystallized, revealing that catalytic activity is associated with protein molecules.
For the 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 important role in gene expression. 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 classical enzymology, and the remainder of this essay, is 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 critical thinking webster catalysing the conversion of nyu essay funny substrate molecules into critical thinking, product molecules as follows: The enormous catalytic activity of enzymes can perhaps best be expressed by 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 a single enzyme molecule per unit time (usually per immigration argumentative essays, minute or per second). Examples of turnover rate values are listed in Table 1. For example, a single molecule of carbonic anhydrase can catalyse the critical thinking definition conversion of over half a million molecules of its substrates, carbon dioxide (CO 2 ) and water (H 2 O), into 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 college depression most a range of similar types) of webster substrate molecule into product molecules. Some enzymes demonstrate group specificity. For example, alkaline phosphatase (an enzyme that is depression, 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 webster, 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. This I Believe Essay By Albert Einstein! glucose) in a complex mixture (e.g. Definition Webster! 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 suffix - in (e.g. trypsin, chymotrypsin, papain). Often the trivial name also indicates the substrate on which the enzyme acts (e.g. glucose oxidase, alcohol dehydrogenase, pyruvate decarboxylase). Nyu Essay Funny! However, some trivial names (e.g. invertase, diastase, catalase) provide little information about the substrate, the product or the reaction involved.
Due to the growing complexity of and inconsistency in the naming of enzymes, the International Union of Biochemistry set up the critical definition Enzyme Commission to address this issue. The first Enzyme Commission Report was published in critical anylisis 1961, and provided a systematic approach to the naming of enzymes. The sixth edition, published in thinking webster 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. For example, the enzyme with the trivial name lactate dehydrogenase has the EC number 220.127.116.11, and is more correctly called l –lactate: NAD + oxidoreductase. The first part of the EC number refers to the reaction that the immigration argumentative essays enzyme catalyses (Table 2).
The remaining digits have different meanings according to the nature of the critical webster reaction identified by the first digit. For example, within the oxidoreductase category, the second digit denotes the college depression hydrogen donor (Table 3) and the third digit denotes the hydrogen acceptor (Table 4). Thus lactate dehydrogenase with the EC number 18.104.22.168 is an oxidoreductase (indicated by the first digit) with the alcohol group of the lactate molecule as the hydrogen donor (second digit) and NAD + as the thinking definition hydrogen acceptor (third digit), and is the 27th enzyme to be categorized within this group (fourth digit). Fortunately, it is now very easy to find this information for admission depression any individual enzyme using the Enzyme Nomenclature Database (available at http://enzyme.expasy.org). Enzyme structure and substrate binding. Amino acid-based enzymes are globular proteins that range in critical thinking definition webster size from this einstein less than 100 to critical definition webster, more than 2 000 amino acid residues. These amino acids can be arranged as one or more polypeptide chains that are folded and bent to 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 constituent amino acids. It is the shape and charge properties of the active site that enable it to bind to a single type of nyu essay funny substrate molecule, so that the enzyme is able to demonstrate considerable specificity in its catalytic activity.
The hypothesis that enzyme specificity results from the critical 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 lock (the enzyme). 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 nyu essay funny this finding, in 1958 Daniel Koshland extended Fischer's ideas and presented the ‘induced-fit model’ of substrate and enzyme binding, in which the enzyme molecule changes its shape slightly to accommodate the binding of the webster 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 i believe essay by albert order to provide a perfect match. Since it is the active site alone that binds to the substrate, it is critical thinking, logical to learning, 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 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 thinking definition webster the enzyme's catalytic activity. A cofactor may be another organic molecule, in to think critically which case it is called a coenzyme, or it may be an inorganic molecule, typically a metal ion such as iron, manganese, cobalt, copper or zinc. A coenzyme that binds tightly and permanently to the protein is generally referred to as the prosthetic group of the critical definition enzyme. When an enzyme requires a cofactor for its activity, the inactive protein component is generally referred to career essays, as an apoenzyme, and critical thinking definition, the apoenzyme plus the cofactor (i.e. the active enzyme) is called a holoenzyme (Figure 2).
The need for minerals and immigration essays, vitamins in the human diet is partly attributable to critical thinking, their roles within metabolism as cofactors and coenzymes. How do enzymes work? The broad answer to this question is that they do not alter the anylisis equilibrium (i.e. the thermodynamics) of a reaction. This is because enzymes do not fundamentally change the critical definition webster 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 chemical equilibrium. In many cases the equilibrium of immigration argumentative essays a reaction is far ‘to the right’—that is, virtually all of the substrate (S) is 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 look at a reaction where the thinking webster equilibrium point is quite central. In this reaction, if we start with a solution of 1 mol l ?1 glucose and essay by albert, add the enzyme, then upon completion we will have a mixture of 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 solution and critical webster, waited many months for the reaction to occur in the absence of the enzyme. Interestingly, we could also have started this reaction with a 1 mol l ?1 fructose solution, and critically, it would have proceeded in critical definition webster the opposite direction until the same equilibrium point had been reached. The equilibrium point for this reaction is learning critically, expressed by the equilibrium constant K eq as follows: Thus for a reaction with central equilibrium, K eq = 1, for 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 critical definition 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 learning critically 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 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 thinking definition, proceed to their equilibrium position. If they did, the reaction would essentially stop (i.e. the career choice forward and critical, reverse reactions would balance each other), and there would be no net flux through the pathway. However, in many complex biochemical pathways some of the individual reaction steps are close to equilibrium, whereas others are far from equilibrium, the argumentative essays latter (catalysed by regulatory enzymes) having the greatest capacity to control the overall flux of materials through the definition webster pathway. Enzymes form complexes with their substrates. We often describe an 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 essay reaction (whatever it might be) is made more favourable. Definition Webster! As soon as the nyu essay funny 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 critical thinking definition webster substrate is converted into an intermediate form (often called the transition state) and then into the product. The exact mechanism whereby the i believe essay by albert enzyme acts to increase the rate of the reaction differs from one system to critical thinking definition webster, another. However, the general principle is that by binding of the substrate to the enzyme, the 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 college admission depression an exergonic reaction a small amount of energy, termed the thinking definition webster activation energy, is needed to learning critically, 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 critical definition, potassium chlorate). When a match burns it releases substantial amounts of choice essays light and heat energy (exergonically reacting with O 2 in the air). However, and perhaps fortunately, a match will not spontaneously ignite, but rather a small input of energy in the form of thinking webster 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 i believe by albert einstein, lower the activation energy of a system by making it energetically easier for the transition state to form.
In the presence of an enzyme catalyst, the formation of the critical thinking transition state is i believe essay by albert einstein, energetically more favourable (i.e. it requires less energy for the ‘kick start’), thereby accelerating the rate at which the reaction will proceed, but not fundamentally changing the energy levels of either the reactant or the critical thinking webster product. Properties and mechanisms of enzyme action. Enzyme kinetics is the career choice study of factors that determine the speed of thinking webster enzyme-catalysed reactions. Career Choice Essays! It utilizes some mathematical equations that can be confusing to students when they first encounter them. However, the critical definition theory of kinetics is both logical and simple, and it is essential to develop an understanding of this subject in order to be able to 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 quick to argumentative essays, perform. In general we would use such discontinuous assays when we know little about the system (and are making preliminary investigations), or alternatively when we know a great deal about the system and critical thinking definition, are certain that the this i believe by albert einstein time interval we are choosing is appropriate. In continuous enzyme assays we would generally study the rate of an thinking, enzyme-catalysed reaction by mixing the enzyme with the substrate and continuously measuring the appearance of immigration essays product over time. Of course we could equally well measure the rate of the thinking definition webster reaction by measuring the disappearance of substrate over time. Apart from the actual direction (one increasing and 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 reaction easy to nyu essay funny, follow using a colorimeter or a spectrophotometer. However, we could in fact use any available analytical equipment that has the capacity to measure the concentration of either the product or the substrate. In almost all cases we would also add a buffer solution to the mixture. Thinking! As we shall see, enzyme activity is nyu essay funny, strongly influenced by pH, so it is 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 thinking definition, 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 immigration, slow down, as shown in Figure 4. A common reason for this slowing down of the speed (rate) of the 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 thinking definition, is denaturing over the course of the experiment, or it could be that the pH of the mixture is changing, as many reactions either consume or release protons.
For these reasons, when we are asked to specify the rate of essays a reaction we do so early on, as soon as the enzyme has been added, and when none of the 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 also quite straightforward, as the critical thinking definition rate is effectively linear, so we can simply draw a straight line and measure the gradient (by dividing the concentration change by the time interval) in order to essays, evaluate the reaction rate over this period. We may now perform a range of similar enzyme assays to evaluate how the initial velocity changes when the substrate or enzyme concentration is altered, or when the pH is changed. These studies will help us to characterize the critical definition properties of the enzyme under study. The relationship between enzyme concentration and the rate of the reaction is essays, 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. Thus there is critical thinking definition webster, 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. Nyu Essay Funny! Initially, when the substrate concentration is increased, the rate of reaction increases considerably. However, as the substrate concentration is increased further the effects on 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 critical thinking enzyme is considered to be coming close to saturation with substrate, and demonstrating its maximal velocity ( V max ). Note that this maximal velocity is in this by albert einstein 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 describe this hyperbolic relationship, just as with a linear relationship ( y = mx + c ), which can be expressed by the two constants m (the slope) and c (the intercept).
We have in fact already defined the constant a — it is V max . Critical Webster! 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 defined as the substrate concentration that gives half-maximal velocity. Our final equation, usually called the nyu essay funny Michaelis–Menten equation, therefore becomes: In 1913, Leonor Michaelis and Maud Menten first showed that it was in fact possible to critical thinking webster, derive this equation mathematically from immigration essays 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 reaction takes place via the formation of an definition, 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 essays 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 product concentration will be negligibly small (i.e. [S] ? [P]), such that we can ignore the thinking definition possibility of argumentative essays any product reverting to webster, 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 rate of formation of product, as the rate at which the immigration argumentative ES complex dissociates to form product.
This is based upon the rate constant k 2 and webster, the concentration of the ES complex, as follows: 1. Since ES is an intermediate, its concentration is unknown, but we can express it in terms of known values. In a steady-state approximation we can assume that although the concentration of admission depression substrate and product changes, the concentration of the ES complex itself remains constant. The rate of formation of the critical ES complex and the rate of 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 substrate greatly exceeds the concentration of critical essay enzyme (i.e. [S] ? [E]), the definition concentration of uncombined substrate [S] is almost equal to the total concentration of substrate. The concentration of uncombined enzyme [E] is equal to the total enzyme concentration [E] T minus that combined with substrate [ES].
Introducing these terms to Equation 3 and solving for ES gives us the 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 maximal velocity. Career! Thus Michaelis and Menten were able to webster, derive their final equation as: A more detailed derivation of the critical Michaelis–Menten equation can be found in 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 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 thinking each substrate.
A low K m value indicates that the enzyme requires only a small amount of substrate in order to become saturated. Therefore the nyu essay funny maximum velocity is 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. Critical Definition! Thus we generally refer to K m as a measure of the affinity of the 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 i believe a particular enzyme. An enzyme with a low K m value relative to the physiological concentration of thinking substrate will probably always be saturated with substrate, and will therefore act at a constant rate, regardless of career choice essays variations in the concentration of substrate within the physiological range. An enzyme with a high K m value relative to 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 definition rate of nyu essay funny formation of product will depend on the availability of substrate. If an thinking webster, 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 essays all cases. If two enzymes (with similar V max ) in different metabolic pathways compete for the same substrate, then if we know the definition webster K m values for learning critically the two enzymes we can predict the relative activity of the critical thinking two pathways.
Essentially the pathway that has the to think enzyme with the critical thinking definition webster lower K m value is likely to be the essay by albert einstein ‘preferred pathway’, and more substrate will flow through that pathway under most conditions. For example, phosphofructokinase (PFK) is the enzyme that catalyses the critical thinking first committed step in college admission depression the glycolytic pathway, which generates energy in critical thinking definition the form of critical 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 critical definition webster K m of PFK for its substrate is lower than that of GUT for its substrate. This Essay! Thus at critical thinking definition webster lower cellular hexose phosphate concentrations, PFK will be active and learning to think critically, 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 the pathway of ATP production, which is the more essential function. Very often it is 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 estimating maximal velocity, and therefore we cannot evaluate half-maximal velocity and thus K m . Fortunately, we can plot our experimental data in critical thinking webster a slightly different way in order to admission essays, obtain these values. Critical Thinking Definition Webster! The most commonly used alternative is the Lineweaver–Burk plot (often called the double-reciprocal plot). This plot linearizes the depression hyperbolic curved relationship, and the line produced is webster, easy to extrapolate, allowing evaluation of V max and K m . For example, if we obtained only the first seven data points in Figure 6, we would have difficulty estimating V max from a direct plot as shown in essays depression 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. a double-reciprocal plot), the thinking webster data are linearized, and essays, the line can be easily extrapolated to the left to provide intercepts on thinking both the y -axis and critical anylisis, 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 critical thinking definition webster the lowest substrate concentrations. Nyu Essay Funny! These concentrations might well be the most prone to error (due to difficulties in 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 thinking definition webster the line of best fit estimated from the data, and therefore on the extrapolated values of both V max and K m . The two sets of 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 a very low substrate concentration and a low reaction rate. However, this single point can have an enormous impact on the line of best fit and the accompanying estimates of 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 the majority of college essays enzymology textbooks, and 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 definition the protein structure. Essay Einstein! The effects of pH and temperature are generally well understood.
Most enzymes have a characteristic optimum pH at which the velocity of the catalysed reaction is maximal, and above and critical thinking definition webster, below which the velocity declines (Figure 9). The pH profile is dependent on a number of factors. As the career pH changes, the thinking definition webster ionization of groups both at the enzyme's active site and on the substrate can alter, influencing the rate of binding of the substrate to the active site. These effects are often reversible. For example, if we take an enzyme with an optimal pH (pH opt ) of this i believe 7.0 and definition webster, 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 enzyme are often restored. However, if we place the enzyme in a more extreme acidic or alkaline environment (e.g. at this i believe einstein pH 1 or 14), although these conditions may not actually lead to changes in the very stable covalent structure of the critical 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 original conformation and nyu essay funny, 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 thinking, 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 career essays opposite directions. Critical Thinking! As the temperature is raised, the rate of molecular movement and hence the rate of reaction increases, but at the same time there is a progressive inactivation caused by denaturation of the admission enzyme protein.
This becomes more pronounced as the temperature increases, so that an critical, 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 college admission protein to a range of temperatures for a fixed period of time, and subsequently measuring its activity at critical webster one favourable temperature (e.g. This By Albert Einstein! 25°C). The temperature at which denaturation becomes important varies from one enzyme to another. Normally it is negligible below 30°C, and starts to 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 critical thinking extremely thermophilic microorganisms, such as thermolysin (a protease from 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 100°C. Enzymes are sensitive to inhibitors. Substances that reduce the anylisis essay activity of an enzyme-catalysed reaction are known as inhibitors.
They act by either directly or indirectly influencing the critical definition catalytic properties of the active site. Inhibitors can be foreign to career choice essays, the cell or natural components of it. Those in the latter category can represent an definition, important element of the regulation of cell metabolism. Many toxins and also many pharmacologically active agents (both illegal drugs and prescription and over-the-counter medicines) act by inhibiting specific enzyme-catalysed processes. Inhibitors are classified as reversible inhibitors when they bind reversibly to an enzyme. A molecule that is this i believe essay by albert einstein, structurally similar to the normal substrate may be able to bind reversibly to critical thinking, the enzyme's active site and therefore act as a competitive inhibitor.
For example, malonate is 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 the enzyme's natural substrate, succinate (see below). When malonate occupies the active site of 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 choice essays reaction). One of the characteristics of 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 thinking certain enzymes, high concentrations of either the substrate or the choice essays product can be inhibitory. Critical Thinking Definition Webster! For example, invertase activity is considerably reduced in the presence of immigration argumentative essays high concentrations of sucrose (its substrate), whereas the ?-galactosidase of Aspergillus niger is 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. Webster! Non-competitive inhibitors react with the enzyme at a site distinct from the active site. Therefore the learning to think 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 their inhibition cannot be overcome by critical definition webster, increasing the substrate concentration. Such inhibitors in effect reduce the career choice concentration of the critical definition webster active enzyme in solution, thereby reducing the V max of the reaction. Critical Anylisis! However, they do not change the value of critical thinking webster K m . Uncompetitive inhibition is rather rare, occurring when the inhibitor is only able to bind to admission essays depression, the enzyme once a substrate molecule has itself bound. As such, inhibition is thinking webster, most significant at high substrate concentrations, and results in a reduction in the V max of the essays reaction. Uncompetitive inhibition also causes a reduction in K m , which seems somewhat counterintuitive as this means that the affinity of the enzyme for critical definition its substrate is actually increased when the essays inhibitor is present.
This effect occurs because the thinking binding of the nyu essay funny inhibitor to definition, 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 observed reaction rates with inhibitor present are always lower than those in the absence of the uncompetitive inhibitor. Irreversible inhibitors and poisons. If an i believe by albert einstein, inhibitor binds permanently to an enzyme it is known as an irreversible inhibitor. Definition! Many irreversible inhibitors are therefore potent toxins. Organophosphorus compounds such as diisopropyl fluorophosphate (DFP) inhibit acetylcholinesterase activity by nyu essay funny, reacting covalently with an important serine residue found within the active site of the enzyme. The physiological effect of this inactivation is interference with neurotransmitter inactivation at the synapses of definition nerves, resulting in the constant propagation of nerve impulses, which can lead to death.
DFP was originally evaluated by learning to think critically, 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 critical thinking webster, the Michaelis–Menten relationship, it is often quite disconcerting to critically, 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 metabolic pathways by responding to inhibitors and activators. These enzymes in fact show a sigmoidal (S-shaped) relationship between reaction rate and 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 area where activity is higher than that of an equivalent ‘normal’ enzyme, with a rapid transition between these two phases. This is 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. Critical Webster! They also have multiple active sites where the immigration argumentative substrate can bind. Much of our understanding of the function of allosteric enzymes comes from critical definition webster studies of haemoglobin which, although it is learning to think critically, not an enzyme, binds oxygen in 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 enzyme and thereby change the definition webster shape of the protein such that the essay remaining active sites are able to definition webster, 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. not at the active site). Molecules that stabilize the protein in 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 argumentative DNA and RNA. The reaction is critical thinking definition webster, 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 nyu essay funny further pyrimidine nucleotides.
As the thinking webster concentration of CTP in nyu essay funny the cell decreases then so does the inhibition of definition ATCase, and argumentative, the resulting increase in enzyme activity leads to critical thinking definition webster, the production of more pyrimidine nucleotides. This negative feedback inhibition is an important element of biochemical homeostasis within the cell. However, in order to synthesize DNA and RNA, the cell requires not only pyrimidine nucleotides but also purine nucleotides, and these are needed in roughly equal proportions. Purine synthesis occurs through a different pathway, but interestingly the final product, the purine nucleotide adenosine triphosphate (ATP), is immigration argumentative, 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 thinking definition, form the polymers DNA and RNA. Thus ATCase is nyu essay funny, able to regulate the production of pyrimidine nucleotides within the cell according to cellular demand, and also to ensure that pyrimidine nucleotide synthesis is synchronized with purine nucleotide synthesis—an elegant biochemical mechanism for critical thinking definition the regulation of an extremely important metabolic process.
There are some rare, although important, cases of monomeric enzymes that have only one substrate-binding site but are capable of critical anylisis essay demonstrating the sigmoidal reaction kinetics characteristic of allosteric enzymes. Particularly noteworthy in this context is the monomeric enzyme glucokinase (also called hexokinase IV), which catalyses the phosphorylation of glucose to critical thinking webster, glucose-6-phosphate (which may then either be metabolized by the glycolytic pathway or be used in glycogen synthesis). Immigration Argumentative Essays! It has been postulated that this kinetic behaviour is a result of individual glucokinase molecules existing in one of critical thinking definition two forms—a low-affinity form and a high-affinity form. Anylisis! The low-affinity form of the enzyme reacts with its substrate (glucose), is then turned into the high-affinity form, and remains in that state for webster a short time before slowly returning to its original low-affinity form (demonstrating a so-called slow transition). 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. Critically! 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 the fact that they catalyse and co-ordinate the complex reactions of cellular metabolism.
Up until the critical 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 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 immigration essays 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 production of rennin (a milk-coagulating enzyme used in cheese manufacture) the traditional approach is to use the critical 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 choice essays a calf's stomach is 10 kg, and it takes several months of critical webster 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. Thus the microbial product is clearly preferable economically, and is free from the ethical issues that surround the use of animals. Indeed, most of the cheese now sold in supermarkets is made from milk coagulated with microbial enzymes (so is suitable for vegetarians).
A further advantage of using microbial enzymes is immigration, their ease of extraction. Many of the webster microbial enzymes used in biotechnological processes are secreted extracellularly, which greatly simplifies their extraction and purification. Critically! Microbial intracellular enzymes are also often easier to obtain than the equivalent animal or plant enzymes, as they generally require fewer extraction and critical thinking definition, 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 career choice essays, 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 critical definition webster which is required.
Finally, enzymes from plant and animal sources show wide variation in yield, and nyu essay funny, may only be available at certain times of critical webster 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 nyu essay funny, plant sources, the stability of microbial enzymes is usually high. For example, the critical thinking definition webster high temperature stability of 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 plants is technically much more difficult, is i believe essay, more expensive and is still the subject of significant ethical concern, especially in the U.K. Enzymes may be intracellular or extracellular. Although many enzymes are retained within the cell, and may be located in critical definition webster specific subcellular compartments, others are released into career choice, the surrounding environment. The majority of enzymes in 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 much smaller amounts by definition, the cell. Examples of 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 depression specific activity) can be determined and used as a measure of enzyme purity. Webster! A variety of methods can be used to remove contaminating material in by albert order to purify the enzyme and increase its specific activity. Definition Webster! Enzymes that are used as diagnostic reagents and in clinical therapeutics are normally prepared to learning to think critically, a high degree of purity, because great emphasis is placed on definition the specificity of the reaction that is being catalysed. Clearly the higher the choice level of purification, the 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 culture broth containing the growth medium, organisms (whole or fragmented) and enzymes of interest.
However, even when the cheapest bulk enzymes are utilized (e.g. proteases for use in washing powders), the enzyme cost can contribute around 5–10% of the critical thinking webster 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 career, stabilize the enzyme product. The pH may also be adjusted to optimize enzyme stability. If the enzyme-producing organism is critical thinking webster, 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 choice, clump and thus come out of critical thinking definition suspension. Extracellular enzymes are found in 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 critically cell wall. • liquid shearing through a small orifice at high pressure (e.g. within a French press) Separation of critical enzymes from the 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 this essay by albert non-protein components by a process of salting out. 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 thinking definition webster solvent is increased by adding an agent such as ammonium sulfate, some of the water molecules will interact with the salt ions, thereby decreasing the this i believe by albert 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 the form of a precipitate. This precipitate (containing the enzyme of interest and other proteins) can then be filtered or centrifuged, and thinking definition, separated from the supernatant. Since different proteins vary in i believe essay by albert einstein the extent to thinking webster, which they interact with water, it is possible to perform this process using a series of additions of ammonium sulfate, increasing the ionic strength in a stepwise fashion and removing the precipitate at each stage. Thus such fractional precipitation is not only admission depression, capable of critical webster separating protein from immigration argumentative non-protein components, but can also enable separation of the 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 critical thinking webster, often effective during the career essays 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 type of thinking webster mobile ion (i.e. cation or anion) it attracts.
Proteins whose net charge is admission, opposite to that of the ion-exchange material will bind to it, whereas all other proteins will pass through the 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 a purification protocol to critical webster, separate molecules on the basis of 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. This By Albert! Separation occurs because the larger protein molecules follow a path down the column between the 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 which a ligand molecule such as a substrate analogue, inhibitor or cofactor of the critical thinking definition 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. Subsequently a solution of the ligand is introduced to the column to release (elute) and immigration essays, thereby recover the bound enzyme from the column in thinking webster 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. Advances in molecular biology enable us to purify recombinant proteins, including enzymes, through affinity tagging. In a typical approach the gene for the enzyme of interest would be modified to code for college admission essays depression a further short amino acid sequence at either the critical thinking webster 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. When a mixture containing the tagged protein of essays 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 column. Elution of the bound protein can then be accomplished by adding imidazole to the column, or by critical thinking definition webster, reducing the pH to critical anylisis, 5-6 to displace the His-tagged protein from the nickel ions. Such techniques are therefore capable of rapidly and highly effectively isolating an enzyme from a complex mixture in only one step, and typically provide protein purities of up to 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 isoelectric focusing.
Enzymes are antigenic, and critical thinking definition webster, since problems occurred in the late 1960s when manufacturing workers exhibited severe allergic responses after breathing enzyme dusts, procedures have now been implemented to choice essays, 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 enzyme with polyethylene glycol and preparing small spheres by atomization) or marumerizing (mixing the thinking 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 college depression much of thinking webster 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 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 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 nyu essay funny malt or grain. The enzyme was primarily used within the textile industry for critical thinking 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 to think the preparation of critical thinking definition 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 critical anylisis essay fermentation industry underwent rapid development as methods for thinking the production of antibiotics were developed. These methods were soon adapted for critical anylisis the production of enzymes. In the 1960s, glucoamylase was introduced as a means of critical thinking hydrolysing starch, replacing acid hydrolysis. Subsequently, in the 1960s and 1970s, proteases were incorporated into essays depression, detergents and then glucose isomerase was introduced to produce sweetening agents in 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 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 enzymes, about critical webster 95% are available from suppliers in essays quantities ranging from critical thinking definition ?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 career choice essays, tonnes per annum).
The global enzyme market is critical definition, currently dominated by the hydrolases, especially the learning critically proteases, together with amylases, cellulases and lipases supplied either as liquid concentrates or as powders or granules that release the soluble enzyme on critical thinking definition webster dissolution. Global production is dominated by two companies, which between them supply more than two-thirds of the global enzyme market, namely the Danish company Novozymes, with a market share of 47%, and the U.S. company DuPont (which recently acquired Genencor), with 21%. The value of the world enzyme market has increased steadily from ?110 million in 1960 to ?200 million in 1970, ?270 million in 1980, ?1 000 million in 1990 and over ?2 000 million in 2010. Food and beverage enzymes represented the 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 essays a further ?700 million. Estimates of future demand are in critical the range of ?4 000–5 000 million between 2015 and 2016, growing at a rate of 6–7% annually. The developing economies of the Asia-Pacific Region, the essays Middle East and critical thinking, Africa are now seen to nyu essay funny, be emerging as the critical thinking definition 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 critically optimum enzyme yield for the process falling somewhere between the optimum biomass yield and the point of maximal enzyme activity within the cells. Relatively small fermenters with a volume of critical thinking definition webster 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 essay which substrates are gradually fed into critical definition webster, the reactor over the course of the fermentation, rather than being provided all at once at 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 the end of the batch run the product must be separated from any unused substrate, and also from the enzyme catalyst. Nyu Essay Funny! 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. Critical Thinking! These processes represent a costly downstream processing stage and generally render the enzyme inactive, so when a new batch run is to be started a fresh batch of enzyme is required. Immobilized enzyme systems, in contrast, ‘fix’ the 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 learning to think critically, mixed with a solution of warm (but not too hot) agar and this is allowed to set, the enzyme will be entrapped (for the purposes of this example let us ignore the fact that the enzyme will gradually leak out of this gel). Critical Definition Webster! 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 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 the next batch run. 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 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 enhanced biocatalyst stability, which may be manifested as: • an increased rate of catalysis.
• prolonged duration of catalysis. • greater operational stability to argumentative essays, extremes of pH, temperature, etc. The particular advantage(s) conferred by critical, immobilization will therefore differ from one system to another. It should be noted that often there may be no biochemical advantage at all, and the simple physical advantage of essays ease of separation of the biocatalyst from the product may be sufficient to favour the thinking webster commercial development of an immobilized process. At this point one problem that will immediately spring to mind for nyu essay funny most students is thinking webster, that they have always been taught to fully mix all of the reagents of a reaction, yet the basic principle of immobilization is to 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 critically, 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 critical thinking definition 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 to think critically, immobilized system has an effectiveness factor of 0.1 (i.e. 10%) and we can reuse the biocatalyst 10 times, we essentially achieve the thinking same overall catalytic activity with both the non-immobilized system and immigration, the immobilized one. However, if we are able to critical thinking definition, reuse the biocatalyst 100 times we in 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 continuous-flow reactors, enabling a continuous supply of substrate to be turned into product as it passes through the to think reactor. The control of 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 held within the critical thinking reactor by a coarse mesh, and this i believe einstein, the product continuously flows out of the reactor as substrate is pumped in. It is thinking definition webster, also possible to i believe essay by albert, produce a packed-bed reactor (PBR) (Figure 13b), in which the agar cubes are packed into a column and the substrate is critical thinking definition, pumped through the bed without any need for stirring. CSTRs and PBRs enable the nyu essay funny enzyme to be reused many times before it needs to be replaced. Critical Definition! For example, in the production of high-fructose syrups, the immobilized glucose isomerase enzyme would typically be used continuously for between 2 and 4 months, and only after this time (when its activity would have dropped to essays, 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 webster 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. Anylisis! They may also result in thinking webster 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 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 critical anylisis equivalent batch processes using soluble enzymes. Critical Thinking Definition Webster! 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 nyu essay funny, not a particularly effective form of critical thinking definition immobilization. The high temperature required to prevent the agar from setting may lead to thermal inactivation of the enzyme, and the agar gel itself is very porous and will allow the enzyme to leak out into nyu essay funny, 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 thinking, a supporting matrix is the oldest method of nyu essay funny immobilization. Critical Thinking Webster! As early as 1916, J.M. Nelson and college, Edward G. Definition Webster! 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 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 critical anylisis, enzyme are simply stirred together for critical definition a time (typically minutes to hours). The binding forces that immobilize the anylisis 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 critical thinking, can lead to desorption of the enzyme at to think a critical time.
However, despite these limitations, such a method was used in critical the first commercial immobilized enzyme application, namely DEAE–Sephadex-immobilized l -amino acid acylase, in 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 career choice essays this material remain positively charged at critical thinking definition pH 6–8 (see Figure 15a) and thus bind strongly to proteins, which are generally negatively charged in nyu essay funny this pH range. If the pH is kept constant, the thinking definition webster 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 process significant economic benefits. Clearly DEAE–Sephadex immobilization is only of value for enzymes that have a neutral-to-alkaline pH optimum. 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 argumentative essays, a more alkaline value. Due to the simplicity and controllability of this immobilization procedure, combined with the economic benefits of reuse of the support, ion-exchange materials are now widely used as the webster method of choice in many industrial settings. Immobilization of enzymes by covalent bonding to activated polymers is nyu essay funny, 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. 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 thinking webster, immobilize a variety of proteins. Activated cellulose supports continue to be popular due to their inherent advantages of high hydrophilicity, ready availability, potential for career choice essays 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 critical thinking reactive group into to think critically, 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 enzyme. An example of thinking webster such a bridging molecule is glutaraldehyde, which contains two aldehyde groups, one at either end of nyu essay funny its (CH 2 ) 3 moiety. At neutral pH values the aldehyde groups will react with free amino groups. Thus one end of the glutaraldehyde molecule may be attached to thinking definition webster, the support, and the other to the enzyme. Covalently immobilized enzymes are strongly bound to their support, so when the proteins denature they are difficult to remove (in contrast to adsorption, as described earlier). Learning To Think! 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 critical thinking definition webster which the immigration support may be reused.
The entrapment of an 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 entrapment techniques is that the enzyme is not in fact attached to anything. Consequently there are none of the steric problems associated with covalent or adsorption methods (i.e. the thinking webster possibility of the enzyme binding in such a way that its active site is obstructed by part of the essays supporting polymer matrix). The example of an enzyme retained in agar, described earlier, is a useful illustration of entrapment. Thinking Definition! A preferable alternative involves mixing the catalyst with sodium alginate gel and extruding this into immigration essays, a solution of calcium chloride to produce solid calcium alginate particles. This technique has the advantage of critical not requiring the use of career essays high temperatures. Critical Webster! However, although it is a popular activity in teaching laboratories, outside that setting it is this i believe essay, 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 thinking definition webster enzyme properties. Earlier in this essay it was suggested that immobilization might change the properties of an enzyme to enhance its stability. Initially it was believed that such enhanced stability resulted from the formation of bonds between the college enzyme and 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 the immobilized state, this is most probably explained by the fact that such proteases in free solution are prone to autodigestion (i.e. Webster! enzyme molecules cleave the peptide bonds of adjacent enzyme molecules), a process that is 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. Consider, for example, immobilization of the enzyme by nyu essay funny, adsorption on definition webster to a polyanionic (negatively charged) support such as cellulose. If the nyu essay funny substrate is a cation (i.e. positively charged), it will be attracted to the support and thus to critical definition webster, the enzyme. In this case the nyu essay funny 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. Thus the microenvironment would also be enriched in critical thinking definition webster H + ions, so the pH surrounding the enzyme would be lower than the career choice pH of the bulk phase. 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 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 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 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 thinking enzyme in career choice the outer part of the particle means that substrate will now diffuse deeper into thinking definition, the particle to learning, reach the previously unused core enzyme molecules. In effect this inner reserve of enzyme will offset the loss of enzyme activity through denaturation, so the system will show little or no overall loss of thinking 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 college essays, natural cell membranes (phospholipid bilayers) within living cells will also probably demonstrate these effects, and immobilized systems thus provide useful models for definition the study of such membrane-bound proteins in living cells. The major industrial processes that utilize immobilized enzymes are listed in Table 7. Sales of immobilized enzymes peaked in 1990, when they accounted for about 20% of all industrial enzyme sales, almost entirely due to immigration argumentative essays, the use of glucose isomerase for the production of sweetening agents. Other commercial applications utilize penicillin acylase, fumarase, ?–galactosidase and amino acid acylase. 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 the use of immobilized enzymes on 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 general public believe that sucrose is responsible for the ‘sweetness’ of food and critical thinking webster, 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 anylisis essay corn (maize). Thinking! Developments in HFCS production have been most prominent in immigration argumentative countries such as the U.S.A., which have a high capacity to produce starch in the form of thinking definition corn, but which do not cultivate significant amounts of sugar cane or sugar beet, and must therefore import either the raw products (for processing) or the 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. These enzymes are cheap and can be used in a soluble form. Since starch has to be extracted from corn at high temperatures (because starch has poor solubility at low temperatures and forms very viscous solutions), the admission essays depression process utilizes enzymes from thermophilic organisms, which have very high temperature optima. Simple corn syrup is therefore composed predominantly of thinking definition glucose, which unfortunately has only this essay, 75% of the sweetness of sucrose. However, in order to make the syrup sweeter the enzyme glucose isomerase, which catalyses the critical thinking following reaction, can be employed: This enzyme (described previously in the section on properties and mechanisms of enzyme action) will produce a roughly 50:50 mixture of glucose and fructose at this by albert einstein 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 critical webster, 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 species of Pseudomona s in 1957, and nyu essay funny, more useful enzymes were isolated throughout the 1960s from species of Bacillus and thinking webster, Streptomyces. Argumentative! 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 critical thinking definition a DEAE ion-exchange support. During the late 1970s, advances in enzymology, process engineering and fractionation technology led to to think, the production of syrups with a higher fructose content, and today HFCS containing 55% fructose is critical definition webster, 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. production of learning HFCS was approximately 8 million metric tons, accounting for 37% of the definition webster U.S. caloric sweetener market, and it is estimated that today about 5% of the entire corn crop in the U.S.A. is used to produce HFCS. Within the anylisis essay dairy industry the production of 1 kg of cheese requires about 10 litres of milk, and produces about 9 litres of whey as a waste product. Whey is critical definition, a yellowish liquid containing 6% dry matter, of which nearly 80% is immigration argumentative essays, lactose. The enzyme lactase (?-galactosidase) may be used to break down lactose to its constituent monosaccharides, namely glucose and definition webster, galactose, which are more soluble than lactose, and 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 most successful commercial process for the treatment of whey.
Using a lactase enzyme obtained from nyu essay funny Aspergillus , immobilized by adsorption and critical definition, cross-linked on to a support resin, whey syrups are produced that have been utilized as an ingredient in drinks, ice cream and this i believe einstein, confectionery products. The Aspergillus enzyme has an acid pH optimum of critical thinking definition webster 3–5, and by operating at low pH the process avoids excessive microbial contamination. Treatment plants that utilize 600-litre columns have been built in Finland, and these are used to treat 80 000 litres of whey per day. Career Choice Essays! This technology has also been used to webster, produce whey syrups in England (by Dairy Crest) and in Norway. Similar technology can also be used to remove lactose from milk. Lactose-free milk is produced for consumption by 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 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 argumentative treating 10 000 litres of milk per day, and was operated at low temperature to 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 webster the years many microbial pathogens have become resistant to depression, natural penicillins, and are now only treatable with semi-synthetic derivatives. These are produced through cleavage of natural penicillin, such that the thinking definition webster G or V side chain is removed from the 6-aminopenicillanic acid (6-APA) nucleus of the essay einstein 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 critical thinking definition, yield 7-aminodesacetoxycephalosporanic acid (7-ADCA), which can then be used to generate a number of immigration argumentative essays important cephalosporin antibiotics (e.g. cephalexin, cephradine, cefadroxil). The development of immobilized penicillin G acylase dates back to research conducted in 1969 by University College London and Beecham Pharmaceuticals in critical webster 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 i believe einstein 1980s and 1990s, world production of penicillins was dominated by European manufacturers, which accounted for definition webster production of around 30 000 tonnes of penicillin per annum, 75% of which was used for career essays the manufacture of semi-synthetic penicillins and cephalosporins. However, over the past 10 years, due to increasing costs of thinking webster labour, energy and raw materials, more bulk manufacturing has moved to immigration, 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. Thinking Definition Webster! However, penicillins and cephalosporins still represent one of the world's major biotechnology markets, with annual sales of nyu essay funny about ?10 000 million, accounting for 65% of the entire global antibiotics market. Enzymes have a wide variety of uses in 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 thinking substances, including drugs, are widespread. Enzymes also play a key role in immunodiagnostics, often being used as the essay agent to amplify the signal—for example, in enzyme-linked immunosorbent assays (ELISAs). Within DNA-fingerprinting technology, the critical webster enzyme DNA polymerase plays a key role in argumentative the amplification of webster DNA molecules in the polymerase chain reaction. However, ‘wet chemistry’ analytical methods are increasingly being replaced by career essays, 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 World Health Organization estimated that over 170 million people had diabetes, and critical thinking, predicted that this figure will rise to over 360 million by 2030. In view of this, many companies have made significant investments in RD programmes that have led to career choice essays, the availability of a wide variety of critical thinking 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 nyu essay funny Yellow Springs Instruments (YSI) model 23A glucose analyser. This device incorporated glucose oxidase together with a peroxide-sensitive electrode to critical thinking webster, measure the hydrogen peroxide (H 2 O 2 ) produced during the following reaction: In this device, the rate of H 2 O 2 formation is a measure of the rate of the reaction, which depends on the concentration of glucose in solution, thus allowing the latter to career choice essays, 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 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 critical definition webster 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 college admission depression, the concentration of glucose in critical definition solution. Over the years the 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 be run, with the essays 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 critical thinking definition sample per analysis. The membrane-bound glucose oxidase typically only needs to be replaced every 3 weeks, thereby reducing the cost of 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 other substances of nyu essay funny biological interest, simply by 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 definition fact technologically more advanced because the career choice enzyme and critical, transducer are more intimately linked on the sensor surface.
The first device of this type was launched in 1986 by Medisense, and was based on technology developed in admission essays the U.K. at definition webster Cranfield and essays, Oxford Universities. The ExacTech blood glucose meter was the size and shape of a pen, and definition webster, used disposable electrode strips. This device was followed by a credit card-style meter in career choice essays 1989. Such devices again rely on glucose oxidase as the biological component, but do not measure the reaction rate via the production (and detection) of thinking definition H 2 O 2 . Instead they rely on direct measurement of the critical anylisis rate of electron flow from glucose to critical thinking definition, 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 essays depression FAD redox centre of glucose oxidase in its oxidized form, and GO x -FADH 2 represents the thinking definition reduced form.
Basically electrons are removed from the glucose molecules and passed via the enzyme to the ferrocene mediator, which then donates them to the working electrode surface, resulting in the generation of an electrical current that is directly proportional to 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 argumentative essays next generation of definition webster sensing technology, and integrated the critical anylisis essay enzyme even more closely with the electrode. Originally developed by Adam Heller at the University of Texas in the 1990s, wired-enzyme electrodes do not rely on a soluble mediator such as the ferrocene used in the Medisense devices. Critical Webster! Instead the enzyme is immobilized in an osmium-based polyvinyl imidazole hydrogel in which the electrons are passed from enzyme to electrode by a series of 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. Learning To Think! Devices of this type are highly amenable to miniaturization. Continuous measuring devices are becoming increasingly available, and may well revolutionize the control of 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 a thin hypodermic needle. This is inserted approximately 5 mm under the skin to measure the thinking definition webster glucose level in the interstitial fluid that flows between the cells. The unit is 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 user to a falling or rising glucose level in time for them to 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 college essays, the intermittent administration of insulin. Therefore, looking to the future, we may confidently expect to see the critical definition 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 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. It is admission essays, 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 thinking cell. Genetic engineering is thus predominantly concerned with modifying the proteins that a cell contains, and genetic defects (in medicine) generally relate to choice essays, the abnormalities that occur in definition the proteins within cells. Much of the molecular age of biochemistry is therefore very much focused on the study of the cell, its enzymes and admission essays depression, other proteins, and 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 critical definition webster, 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 career Technology.
For further information and to provide feedback on critical thinking webster 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 open access article published by Portland Press Limited and distributed under the Creative Commons Attribution Licence 3.0.
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Critical thinking Definition in the Cambridge English Dictionary
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How the problem is perceived and understood greatly affects how it can be solved or minimized. Solutions are significantly dependent on the level of understanding of the this i believe essay, problem. To cut it short, if the people do not know how to thinking, define quality life and how to maintain it then it will be hard for them to suggest for practical solutions to the problem. Thus; it is reasonable to say that solutions and policies must be based on the people's perception or suggestion meeting what experts think is the this essay by albert, best. So, policy-makers must consult both suggesting bodies before imposing a policy to critical, solve the problem. Murder and everyday traffic are just two of the most common social problems alive today. From those social problems you can already write the best college essays. Nyu Essay Funny. But, there are other conditions that few people consider as social problems but to some are not. One example is critical definition webster sound pollution among teenagers who play loud party music in essay by albert einstein public places.
To some, this is an undesirable condition but to critical thinking definition, young people, it is their way of critical anylisis expressing themselves. Critical. For nonsmokers, smoking is a bad social condition and should be banned, especially in nyu essay funny public places. Almost all newspapers are filled with different stories and essay on social problems categorized by people as social problems. Drug abuse, crime, abortion, and environmental problems are simply the many conditions. Social problems are always there. Definition. In every state, national or international levels undesirable social conditions are always present. Essays. You may check on the Public Policy Analyst to see some of the existing social problems in the community. Take the specific community location. A typical community usually composed of thinking webster school, school district, town, village and city (some even have best resume writing service agencies). Some have the county.
But, there are social problems which can exist on all the said community locations. Say, for example, there is a rampant case of theft in the whole school district. Learning To Think Critically. So police agencies make surveys and statistics to monitor the crime rate or the existence of a social problem for a specified location under their jurisdiction. Describing your social problem can be done by considering the geographical setting before going into courseworks. Or, you may state the name of the place itself or schools like Bronx High School or New York City. However, PPA is only good for local problems and not on the national level. However, what makes essays on social problems controversial is how we address the word public or people.
When can we say that it is the public that states the critical webster, social condition? How many can be considered as public or the majority and, how few is the minority? This can be traced by einstein using a specific methodology for determining social problems. It is like doing a case study method. Besides the question of how to identify the public, we may also want to consider how to thinking webster, determine the overall social problem. Anylisis Essay. It is not reasonable to critical definition, simply conduct a survey and leave all the responsibility to the people. Critical Anylisis Essay. It is also unreasonable to put the entire load to critical, the experts. Learning Critically. One medium to help us examine the current social problems in the society is mass media, which is also a big help in a research proposal.
The television for example, through coverage, the issue it chooses to present gives a very powerful impact to the public. A majority of the stereotypes that societies live by are brought or influenced by the media today. The effect of critical webster mass media to the society is remarkable (this can be well noted if you examine an essay on social problems in to think India). This explains why the media itself is a good source of determining the major social problem that society has. How the presented information is gladly taken by the public because it is by far the most influential and most effective medium of sharing information to people of all ages. This is why it is both helpful and critical thinking webster, dangerous because children or minor age can already grasp or see contents only i believe essay einstein, intended for adults. Getting heavy and harsh contents for a young age is quite dangerous to the normal development of thinking a child’s psychological health.
Media may not only argumentative essays, comprise of television and social networks. Print ads and magazines are two of the most influential media outlets that you should consider too. If you are not convinced about the power of mass media, check the critical thinking definition, other essay on social problem. Essay On Social Problems: Giving solution to the problem. So, in to think critically order to answer questions on the society’s social problems, we can refer it on how we define the quality of life every individual should have and how it should be deployed. Both public opinion and experts' point of view can be used to carefully define social problems. So, the quality of life according to public opinion should coincide with how professionals and experts identify it.
At what condition that the quality of life is put to critical definition webster, a risk? Up to what extent such condition is critical essay tolerable so as not to decrease the quality of life? How people perceive a situation can be very different from how experts see it. So, it is of great importance that these two bodies coincide with each other. In short, there is a need to study and to understand people's perception as well as their desires and thinking definition webster, needs. How people perceive a situation is greatly influenced by their judgments and their present condition or how they are affected from what they see, hear and essay, feel. So, they normally perceive a situation as threatening if they feel that their necessities are put to risk.
Necessities do include their personal security, health and family’s well-being. Quite simply, if the situation or condition affects the quality of life they perceive and the condition deprives them of webster their necessities then we have a social problem. By Albert. How the problem is perceived and understood greatly affects how it can be solved or minimized. Solutions are significantly dependent on the level of understanding of the problem. To cut it short, if the thinking, people do not know how to define quality life and how to maintain it then it will be hard for them to suggest for practical solutions to learning to think, the problem. Thus; it is reasonable to say that solutions and policies must be based on the people's perception or suggestion meeting what experts think is the critical thinking definition, best.
So, policy-makers must consult both suggesting bodies before imposing a policy to solve the problem.