2020-1 Enzyme Engineering     Problem set #3 (by May 21, 5pm)

1. The following data were recorded for the enzyme-catalyzed reaction.

Substrate concentration (M)	Reaction velocity (nM/min)
6.25 x 10-6	15
7.50 x 10-5	56
1.00 x 10-4	60
1.00 x 10-3	75

(1) Estimate Km and Vmax.

(2) What would V be at S=2.5 x 10-5 ?

2. The following results were obtained from an enzyme reaction. Calculate the kinetic parameters (Km and Vmax) in this reaction. You can determine initial reaction velocity for each substrate concentration by the plots, time (X-axis) vs. product formed (Y-axis).

Substrate concentration (mM)	Product formed (mM) at each time (t, min)
	t=0.5	t=1	t=1.5	t=2	t=2.5	t=3
1.5	3.3	5.6	7.9	10.2	12.2	13.8
2	3.6	6.3	8.9	11.6	13.8	15.4
2.5	4.0	6.9	9.9	12.8	15.0	16.8
3.33	4.3	7.5	10.8	14.0	16.3	17.5
5	4.7	8.4	12.1	15.8	17.7	18.4
10	5.2	9.5	13.7	18.0	19.2	20.0