S. a) Consider the enzymes in the table below and answer the following questions
ID: 1031508 • Letter: S
Question
S. a) Consider the enzymes in the table below and answer the following questions. Answers should be brief (1-3 sentences (6+4 pts) Enzyme Km (M)Kse 95x10 1.4x10 3.5x102 1.0x107 4.5x10 7.5x102 i) Which enzyme has the highest affinity for substrate? How do you know? ii) Which enzyme converts the most substrate to product in a given period of time? How do you know? iii) Which enzyme has the highest catalytic efficiency? How do you know? b) You want to load 15 g of protein in 10 L into one of the 12% polyacrylamide gel well The protein needs to be in lX buffer and in a total volume of 0.100 ml. You are given a 5 mg/ml protein solution, a 20X sample buffer, and distilled water. How much of each would you mix together to make required volume?Explanation / Answer
1) Km also known as Michaelis Menten constant is the measure of the substrate concentration at half maximal velocity. The lower the Km of an enzyme the lower the is the substrate concentration required to saturate the enzyme completely. This means the enzyme which has lower Km has higher affinity toward substrate and can bind the substrate strongly even at low concentration.
Thus the Enzyme C has the highest affinity for substrate as it has lowest Km.
2) Kcat is also known as turnover number, and it is the measure of number of substrate molecules converted to product per enzyme site per unit time. Higher the Kcat higher is the number of substrate converted to product per unit time.
so accordingly Enzyme B has highest Kcat and it will convert the most substrate to product in a given period of time.
3). Catalytic efficiency is calculated by the following formula.
Efficiency = Kcat/Km .
Efficiency of enzyme A = 1.4* 10^4/ 9.5* 10^-5
Efficiency of enzyme B = 1.0* 10^7/ 3.5 * 10^-2
Efficiency of enzyme C = 7.5* 10^2/ 4.5* 10^-6 Enzyme c is the highest catalytic efficiency
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