You are the quality control engineer for an industrial enzyme company. For a bat

Question

You are the quality control engineer for an industrial enzyme company. For a batch of a new enzyme preparation you do some kinetics experiments to make sure that the activity is within specifications, by testing preparations of 1 mg/L of the protein in the presence of various concentrations of the substrate (data in table below). Assuming that the enzyme operates according to uninhibited Michaelis-Menten kinetics, are you at least 95% confident that this batch of enzyme meets specifications of 2000 units mg-1 of protein (where 1 “unit” is equivalent to the maximum conversion rate of 1 micromol of substrate per minute)?

[S] mM

0.8

1.6

3.2

6.4

12.8

25.6

50

v (mM min-1)

0.596

0.913

1.254

1.629

1.784

1.904

1.972

Explanation / Answer

OBSERVATIONS

[S] (uM)

V (mM min-1)

1

0.8

0.596

2

1.6

0.913

3

3.2

1.254

4

6.4

1.629

5

12.8

1.784

6

25.6

1.904

7

50

1.972

Estimated Vmax= Kcat X [Etotal] = (2000 unit mg-1 min-1) X (1 mg ) = 2000 unit min-1 = 2000 umoles min-1

From the table, it is evident that Vmax for the enzyme is approximately 1.972 mM min-1 or 1972 umoles min-1.

Hence, it can be concluded that this batch of enzyme meets specifications of 2000 units mg-1 of protein with 95% confidence.

OBSERVATIONS

[S] (uM)

V (mM min-1)

1

0.8

0.596

2

1.6

0.913

3

3.2

1.254

4

6.4

1.629

5

12.8

1.784

6

25.6

1.904

7

50

1.972

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