Penicillin is hydrolyzed and thereby rendered inactive by penicillinase, an enzyme present in some resistant bacteria.  The mass of this enzyme in Staphylococcus aureus is 29.6 kd.  The amount of penicillin hydrolyzed in 1 minute in 10 ml solution containing 10–9 g of purified penicillinase was measured as a function of the concentration of penicillin. Assume that the concentration of penicillin does not change during the assay. [Penicillin] µM            Amount hydrolyzed (nanomoles) 1                                  0.11 3                                  0.25 5                                  0.34 10                                0.45 30                                0.58 50                                0.61   a) Plot V0 versus [S] and 1/V0 1/[S]. Does penicillinase appear to obey Michaelis-Menten kinetics?   b) What is the value of KM?   c) What is the value of Vmax?

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Penicillin is hydrolyzed and thereby rendered inactive by penicillinase, an enzyme present in some resistant bacteria.  The mass of this enzyme in Staphylococcus aureus is 29.6 kd.  The amount of penicillin hydrolyzed in 1 minute in 10 ml solution containing 10–9 g of purified penicillinase was measured as a function of the concentration of penicillin. Assume that the concentration of penicillin does not change during the assay.

[Penicillin] µM            Amount hydrolyzed (nanomoles)

1                                  0.11

3                                  0.25

5                                  0.34

10                                0.45

30                                0.58

50                                0.61

 

a) Plot V0 versus [S] and 1/V1/[S]. Does penicillinase appear to obey Michaelis-Menten kinetics?

 

b) What is the value of KM?

 

c) What is the value of Vmax?

 

d) What is the turnover number of penicillinase under these conditions?

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