4. Penicillin is hydrolyzed and thereby rendered inactive by penicillinase (also

Question

4. Penicillin is hydrolyzed and thereby rendered inactive by penicillinase (also known as - lactamase), an enzyme present in some penicillin-resistant bacteria. The mass of this enzyme in S.aureus is 29.6kDa. The amount of penicillin hydrolyzed in 1 minute in a 10-mL solution containing 10 9g of purified penicillinase was measured as a function of the concentration of containing 103g of purified penicillinase was m penicillin. Assume that the concentration of penicillin does not change appreciably during the assay [Penicillin] uM Amount hydrolyzed (nmol) 0.11 0.25 0.34 0.45 0.58 0.61 3 30 50 a) Plot these data and determine the Km and Vmax, if the data appear to obey Michalis-Menten kinetics b) What is the turnover number of this enzyme? Assume there is only one active site per enzyme

Explanation / Answer

According to Lineweaver-burk plot

1/V= (KM/Vmax)*(1/S)+ 1/Vmax

where V= Rate of hydrolyzation = moles hydrolyzed/unit time, KM, Vmax are constants and Vmax= maximum rate

the intercept is 1/Vmax and slope is KM/Vmax

The data points along with the plot are shown below.

from the plot , intercept is 1/Vmax= 1*109, Vmax= 10-9 M/s amd slope KM/Vmax= 7651, KM= 7651*10-9 M

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