-->

Pages

Thursday, March 15, 2012

The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant

If you order your cheap term papers from our custom writing service you will receive a perfectly written assignment on The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant. What we need from you is to provide us with your detailed paper instructions for our experienced writers to follow all of your specific writing requirements. Specify your order details, state the exact number of pages required and our custom writing professionals will deliver the best quality The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant paper right on time.

Out staff of freelance writers includes over 120 experts proficient in The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant, therefore you can rest assured that your assignment will be handled by only top rated specialists. Order your The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant paper at affordable prices with cheap essay writing service!



The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction Evaluation Of The Michaelis Constant


Chemical reactions in living organisms occur rapidly at moderate temperatures and under mild conditions primarily because of the catalytic action of specialized proteins called enzymes. Every step in the biological processes are controlled by Enzymes. The lack of even one enzyme can prove fatal. The actual catalytic site of an enzyme molecule is a small area where the enzymes shape is arranged to precisely accept its substrate into this receptor site. The substrate is the substance acted upon by the enzyme and changed to the product. The rate of an enzyme-catalyzed reaction depends partly on how quickly the enzyme can catalyse the substrate, and also on how quickly they are able to join together. Therefore an environmental factor (such as pH or high temperature) which might alter the shape of either enzyme or substrate could alter the rate of reaction. In addition the concentration of active enzyme and substrate molecules would be expected to have important effects on the rate. The rate of an enzyme-catalyzed reaction may be measured by (1) the disappearance of substrate, or () the appearance of product. The enzyme and substrate are mixed and allowed to react for a certain time period; the amount of either the substrate or the product is then measured. This amount gives the rate of activity of the enzyme per unit of time. Alkaline Phosphatase is the enzyme used in this experiment, and the results that will be taken will measure the amount of product produced by measuring its absorbance. Also in each test tube there will be different amounts of substrate so the Michaelis � Menton constant can be worked out.


The Michaelis constant is an approximate measure of the affinity of the enzyme to its substrate. It can be shown as a curve graph, as shown below.





Cheap Custom Essays on The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant




In most cases the curve is a rectangular hyperbola which can be described by the general equation-


Vmax = Km + 1 where Vmax and Km are constants


v S


The curve above is known as a Michaelis - Menton plot. The equation below it is the Michaelis - Menton equation, where Km is known as the Michaelis constant. It has a value that is indicidual for each and every enzyme with a particular substrate, and has units of substrate concentration, ie Molarity.


In general the results can be plotted more accurately in straight line form therefore linear derivations have been obtained for the Michaelis � Menton equation.





The enzyme used is Alkaline Phosphatase, which is usually found in the bone, liver, intestines and kidney. It acts upon phosphoric esters with the liberation of inorganic phosphate, as shown below in the diagram.





The substrate that will be used as the phosphoric ester is Disodium Phenyl Phosphate, which when acted upon by the enzyme is converted into Phenol, and Phosphoric Acid, as shown below in the diagram.





It is not possible to monitor the reactants directly, but if the phenol produced is reacted with 4-aminophenazone, to give a condensation product, which is then oxidized by potassium ferricyanide it gives a coloured quinone, whose absorbance can be measured. This process can be shown below.





In order to measure the rate of enzyme activity converting the substrate into product must be quantified. Some method of measuring the change is necessary and it is more accurate to measure the increase in product from 0, rather than to measure a decrease in substrate from a large amount. In order to keep the incubation time the same in this case, the enzyme was added to the substrate at timed intervals, incubated for 10 minutes and then stopped, with 0 seconds in between each test tube, so at time ) the enzyme is added to the first test tube, at 0.0 the enzyme is added to the second test tube, and so on. To stop the reaction sodium hydroxide was added, for instance at time 10.00 it was added to test tube one, and at 10.0 it was added to test tube , and so on till all test tubes have been started and stopped in this method, which is known as a discontinuous assay.





Results


1 4 5 6 7 8 10 11 1 1


cm


0.1M Buffer pH10 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0


Distilled Water 1. 1. 1.7 1.7 1.5 1.5 1. 1. 1.1 1.1 0. 0. 0.


0.015M Disodium Pheyl Phosphate 0 0 0. 0. 0.4 0.4 0.6 0.6 0.8 0.8 1.0 1.0 1.0


Equilibrate at 7°C for 10 minutes


0.4M NaOH - - - - - - - - - - - - 1.0


Alk. Phosphatase 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1


Incubate at 7°C for 10 minutes


0.4M NaOH 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 -


0.6M NaHCO 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0


6% Aminophenazone 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0


.4% Potassium Ferricyanide 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0


Read Absorbance At 510nm


Absorbance at 510nm 0.00 0.00 0.057 0.05 0.070 0.074 0.0 0.06 0.07 0.08 0.10 0.07 0.00








Tube Number Substrate (s) mM Reaction velocity (v) ยต









































Please note that this sample paper on The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant is for your review only. In order to eliminate any of the plagiarism issues, it is highly recommended that you do not use it for you own writing purposes. In case you experience difficulties with writing a well structured and accurately composed paper on The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant, we are here to assist you. Your cheap custom college papers on The Effect Of Substrate Concentration Upon The Rate Of An Enzyme Catalysed Reaction: Evaluation Of The Michaelis Constant will be written from scratch, so you do not have to worry about its originality.

Order your authentic assignment from cheap essay writing service and you will be amazed at how easy it is to complete a quality custom paper within the shortest time possible!



No comments:

Post a Comment

Note: Only a member of this blog may post a comment.