Lab 3 - Enzyme Kinetics

Lab 3 - Enzyme Kinetics - Enzyme Kinetics INTRODUCTION:...

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Enzyme Kinetics INTRODUCTION: Enzymes can bind either competitively or uncompetitive, we well be finding the rate at which an uncompetitive inhibitor such as tyronsinase will bind to the substrate DOPA. PRINCIPLE: Through enzymes chemical reactions within the body occur at a certain rate. In order to find the rate at which an enzyme binds to a substrate the following formula has been given: E + S k 2 ← k 1 → ES k 4 ← k 3 → E + P . This formula tells the rate at which an enzyme binds to a substrate to give a enzyme substrate, and then tells the rate at which the enzyme substrate complex converts to the enzyme and product. In this experiment we will be finding the rate at which the enzyme binds to the substrate to form an enzyme substrate complex. In order to do that a UV-VIS spectrophotometer is being used. The UV-VIS is able to show the rate of the enzymatic reaction because it measures the amount of diffracted light going through a sample. The amount of diffracted light is monitored and the spectrophotometer shows the amount of diffracted light in the form of absorbance. Using the absorbance researchers can calculate the concentration of the sample and the rate at which the enzyme is binding to a substrate. This data is plotted through a Michealis-Menton plot, this plot is important because it shows the initial reaction velocity versus the substrate concentration. In addition, this plot shows the point at which all the enzyme active sites are filled with substrate molecules, this portion is called V max . Moreover, this plot shows information about the enzyme’s rate of forward and reverse rate or the Michaelis constant (K m ). This constant shows the concentration that leads to the initial reaction velocity and the dissociation constant of the ES complex. 1 From the Michealis-Menton plot, the inverse values of the substrate concentration and initial velocity are taken, also known as Lineweaver-Burk, this plot shows what kind of inhibition occurs with the addition of these enzymes. From our experiment, we expect to see that the oxidation L-DOPA and tyrosine is a production of non-competitive inhibition.
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Lab 3 - Enzyme Kinetics - Enzyme Kinetics INTRODUCTION:...

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