IV. Experiment 2 - Effects of Varying Substrate Concentration In this experiment
ID: 207803 • Letter: I
Question
IV. Experiment 2 - Effects of Varying Substrate Concentration In this experiment, the amount of enzyme will not be altered but the substrate concentration will be varied. (10). Hypothesis: Propose a hypothesis about the effects of varying substrate concentration on product formation when the amount of enzyme is held constant a. Ho: b. Ha: Procedure: 1. For this experiment, choose the optimal enzyme dilution from experiment 1 (choose dilution that did not turn yellow too quickly but eventually did turn a bright yellow in the time frame). This enzyme dilution will be used for the following experiment. It may be necessary to prepare more of the dilution. (11). How will you go about preparing more of the chosen dilution? 2. Prepare a serial dilution of substrate (ONPG) by obtaining four cuvettes and following the procedure in the table below. The stock solution is 5mM. Fill in the % dilutions.Explanation / Answer
The enzymes are the molecules that are used to catalyze a reaction in various different biological processes. In the experiment, this enzyme used must not be changed and the substrate concentration only must be altered. The effects of varying substrate concentration are to be shown. The substrate concentration is quite important for determining the rate of a reaction.
As we know that the enzyme is kept constant, the substrate concentration is changed and the effects on the product formation are noted.
a. Ho (The hypothesis which is generally assumed to be true): The substrate concentration is started from a small value and increased gradually. The enzyme is kept constant. As the reaction goes on, the rate also remains same and the product formation is not much effect at different concentrations. There can be a linear relationship between the rate of reaction or the rate of product formation and the substrate concentration.
b. Ha (The hypothesis, which contradicts the null hypothesis): If the enzyme is kept constant, it will be abundantly available in low concentrations of the substrate. However, as we increase the substrate concentration, the enzyme gets used up and then it will not be available for all the substrate molecules together.
Therefore, as we increase the substrate concentration, initially the rate of reaction increases. But, as the concentration is increased beyond a point, the enzyme gets decreased and the rate of reaction attains a constant. So, there is a hyperbolic relationship between the rate of reaction and the substrate concentration.
So, the product formation is quick initially and the rate is faster and then decreases gradually as the concentration of the substrate is increased.
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