Eenzyme reaction where substrate reduced to ethanol

Reference no: EM1396732

You are following an enzyme reaction where a substrate (Acetaldehyde) is reduced to Ethanol using NADH as a source of reductant, and releasing NAD+. The loss of NADH can be followed by tracking the loss of absorbance at 340 nm based on the extinction coefficient of ? = 6220 M-1 cm-1. You place the following materials in a cuvette; 5 ?g of substrate, 50 ?L of enzyme (1:10 dilution of 0.4 mg/mL stock enzyme), and NADH to bring the concentration to 250 ?M all in a final volume of 1 mL with a cuvette with a pathlength of 1 cm, and find an initial loss in absorbance at 340 nm of 0.063 per minute. Assuming the substrate is well above the KM of this enzyme, what is the specific activity of the enzyme (in mols min-1 (mg protein)-1) based on your results?

Reference no: EM1396732

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