Penicillin is hydrolyzed and thereby rendered inactive by penicillinase (also known as _lactamase),

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Penicillin is hydrolyzed and thereby rendered inactive by penicillinase (also known as _lactamase),

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Penicillin is hydrolyzed and thereby rendered inactive by
penicillinase (also known as _-lactamase), 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 a 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 appreciably during the assay. [Note that kilodalton is the unit
for MW for biological molecules. A dalton is another name for atomic mass unit.
For

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Penicillin is hydrolyzed and thereby rendered inactive by
penicillinase (also known as _-lactamase), 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 a 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 appreciably during the assay. [Note that kilodalton is the unit
for MW for biological molecules. A dalton is another name for atomic mass unit.
For example, a protein with a mass of 15 kd means that the MW is 15,000 g/mol.]

a. Plot V0 versus [S] for these data. Does penicillinase
appear to obey MichaelisMenten kinetics? If so, what is the value of Km?

b. What is the value of Vmax? Indicate on the plot how you
determined Km and Vmax.

c. Write the MichaelisMenten equation and show the
derivation of the LineweaverBurke equation.

d. Plot 1/V0 versus 1/[S].

e. Determine Km and Vmax using the LineweaverBurke plot.
Indicate on the plot how you determined Km and Vmax.

f. Assume there is one active site per enzyme molecule. What
is the turnover number of penicillinase under these experimental conditions?

g. How much time does it take for the penicillinase to
hydrolyze one penicillin molecule?

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