s) In two experiments carried out in a research lab, an enzyme reaction is run with and without an inhibitor. Samples were taken over time to measure the concentration of product as the reaction proceeds, with raw data given below. Data were collected at 34 °C and pH 7.5, using [S] = 0.115 M (for both trials) & [E]. = 0.0024 M (for both trials): Product concentration (mol P/L) No inhibitor [I] = 0.019 M 0 0 Time min 0 3.5 9 13 17 26 43 69 104 156 208 311 415 519 0.0016 0.0038 0.0052 0.0078 0.0111 0.0180 0.0281 0.0412 0.0525 0.0587 0.0598 0.0600 0.0600 0.0011 0.0023 0.0031 0.0045 0.0074 0.0119 0.0195 0.0271 0.0410 0.0479 0.0550 0.0588 0.0600 A. Determine the initial reaction velocities for both reactions. 3. From these data, can you determine what type of inhibition is occurring? If so, show evidence of the type of inhibition occurring. If not, describe what experiment(s) should be done to determine the type of inhibition (competitive, non-competitive or uncompetitive), as well as how the data should be treated to find the inhibitor type.

From a kinetics experiment, kcat was determined to be 295sec-1.  For the kinetic assay, 0.3mL of a 0.25mg/mL solution of enzyme was used, and the enzyme has a molecular weight of 125,000 g/mole.  Assume a reaction volume of 3mL.  Calculate Vmax (µM∙min-1) for the enzyme and catalytic efficiency ( in M-1sec-1) for the enzyme. The Km for the enzyme was determined to be 2.55 x 10-2M.

The enzyme, fumarate, has the following kinetic constants: k 1 k 2 k -1 where k 1 = 10 9 M -1 s -1 k -1 =4.4 x 10 4 s -1 k 2 = 10 3 s -1 a. What is the value of the Michaelis constant for this enzyme? b. At an enzyme concentration of 10 -6 M, what will be the initial rate of product"

Draw the Michaelis-Menten graph you would predict for the enzyme using 50 nM and 100 nM of enzyme in your assays.