You run an enzyme assay designed to determine the alpha-glucosidase activity aft

Question

You run an enzyme assay designed to determine the alpha-glucosidase activity after 1 hour (60 min) by determining the concentration of p-nitrophenol. The results you collected are presented below. Enzyme Dry Weight Absorbance at 580 nm: 0.220 The Required dilution factor: 60 p-nitrophenol absorbance at 400 nm: 0.592 You now need to analyze the results and calculate the following: (Answer each question to 3 significant figures) Calculate the Dry Weight (DW) of your enzyme using the following equation. DW_enzyme = 0.35. A_580. D_F = g If the molar absorptivity of the p-nitrophenol is 7.28 mM^-1 cm^-1 and the path length is 1.0 cm, what is the concentration of p-nitrophenol in the sample? mM Using the formula below, what is the activity of your enzyme? Activity = A_400 * 1000/0.00728 * DW_enzyme * 60 = (mu mol/min.g)

Explanation / Answer

Solution:

DWenzyme = 0.35 * 0.220 * 60 = 4.62 gm

Cp-nitrophenol = A400/e*l = 0.592 / (7.28*1) = 0.081318 mM

Activity = (0.592 * 1000) / (0.00728 * 4.62 * 60) = 293.3574

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