Question

6. You have been working with the jarase enzyme which is not well understood. The jarase enzyme catalyzes the reaction shown below between B-hydroxy-2-butanone (B-H2b) and NAD'. You have noticed that jarase cannot catalyze the reverse reaction: OH + NAD NADH + H You generated the following kinetic data for the jarase enzyme, all reactions are carried out under the same reaction conditions, which are: pH 7.0, 1 mM NAD' varying amounts of B-H2b (from 0.002 - 2 mm) and 0.1 MM jarase. You tested the activity of the jarase enzyme and generated the following results: Jarase enzyme Test molecule KM (mm) cat (min) Wild type 0.31 0.08 947 100 Wild type 2 mM U41 0.25 0.06 Wild type 2 mM U42 0.1 0.006 316 60 C132A 1.7 + 0.2 891 : 100 C132A 2 mM U41 2.0 10.5 39 12 Y35F 0.32 0.08 903 90 Y35F 2 mM U41 0.26 +0.08 723 + 150 Use this information to answer questions 6a - 6i. 6a. What class of enzymes would you place jarase into based on the information provided? (1 point) 66. Does the reaction carried out in these experiments adhere to the steady state assumption Yes or no, and explain why or why not. (3 points) 6. What is NAD considered to be in this reaction? (2 points)

Answer 1

Ans-6a the enzymes will be grouped as refuctase or tranferase as it was used 6o reduce and transfer a group.

Ans-6b no the reaction is not is steady state as the data is indicating a great variation in the Km and Kcat.

Ans-6c the NAD is used to accept the H ions from the substrate. It will form a by product.