Bio I Lab Report - Effects of Competitive Inhibitors on Enzyme/Substrate Reactions Christina Carr 2:10:08 October 9 2007 Ukpong Eyo For the

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Effects of Competitive Inhibitors on Enzyme/Substrate Reactions Christina Carr 2:10:08 October 9, 2007 Ukpong Eyo
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For the experiment regarding competitive inhibition we were to make four different mixtures using frozen beef heart. The four test tubes were to contain a certain mixture of beef heart, Malonate(inhibitor), Succinate(substrate), Phosphate buffer, water, and Methylene blue.(Frankel, Williams, Irish, and Stefaniak pg. 34). After creating the mixture we sealed the tube with paraffin oil and placed the tubes in a water bath at 37ºC for half an hour. After the half an hour we removed the test tubes and recorded our results. This experiment was to show a real example of competitive inhibition. When the beef heart is mixed with Succinate the substrate the protein is broken down into amino acids. When Malonate is added, it should inhibit the break down of the proteins into amino acids. Or in other words it binds to the enzyme blocking the substrate from attaching to the enzyme which triggers the breakdown of proteins into amino acids. For the test tube containing the substrate, enzyme and the inhibitor, tube number three, it was predicted that there should be some form of reaction, but not a complete reaction. The Malonate should inhibit some of the substrate from attaching to the enzyme but not all of them. When the protein is broken down into amino acids the reaction causes the Methyl blue in the solution to change from blue to colorless. Therefore when there is a substrate and an enzyme present without an inhibitor the solution should be colorless when removed from the water bath as in tube number two. If the substrate is not present or the enzyme is not present the solution should not change in color and therefore should be blue when taken out of the water bath. This is shown in tubes number one and four. When an inhibitor is present along with a substrate and an enzyme there should not be as
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obvious of a reaction as without an inhibitor but there should be a slight color change, somewhere between bright blue and colorless. The first step of the experiment was to collect the beef heart into the four test
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This note was uploaded on 03/30/2008 for the course BIO 002:010 taught by Professor Denburg during the Fall '08 term at University of Iowa.

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Bio I Lab Report - Effects of Competitive Inhibitors on Enzyme/Substrate Reactions Christina Carr 2:10:08 October 9 2007 Ukpong Eyo For the

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