A research group discovers a new version of happyase, which they call happyase*,

Question

A research group discovers a new version of happyase, which they call happyase*, that catalyzes the chemical reaction HAPPY SAD The researchers begin to characterize the enzyme (a) In the first experiment, with [EJ at 8 nM, they find(b) In another experiment, with [EJ at 3 nM and b In another experiment, with [Et] at 3 nM and that the Vmax is 3.08 IM s-1 Based on this experiment, what is the [HAPPY] at 40 M, the researchers find that Vo 774 nM s-1 What is the measured Km of happyase* for its substrate HAPPY? kcat for happyase*? Number Number cat UM Im c) Further research shows that the purified happyase"used in the first two experiments was actually contaminated with a reversible inhibitor called ANGER. When ANGER is carefully removed from the happyase* preparation, and the two experiments repeated, the measured Vmax in (a) is increased to 7.70 iM s-1 and the measured Km in (b) is now 22.4 M. For the inhibitor ANGER, calculate the values of ? and ?. Number Number

Explanation / Answer

a. Convert mM and nM into Moles
Kcat = Vmax / [Et] = 3.08 X 10-6 / 8 X 10-9

= 385 s-1

b. Vo = Vmax / 2

Vmax = 1548 X10-3

Calculating Km using michaelis menten eq.

Vo = Vmax [S] / Km + [S]

774 X 10-3 = [(1548 X 10-3) X 40] / Km + 40

Km = 40 uM

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