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 kDa. 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 kDa means that the MW is 15,000 g/mo
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 kDa. 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
Penicillin (M) Amount hydrolyzed (nanomoles)
1 0.11
3 0.25
5 0.34
10 0.45
30 0.58
50 0.61
4. Plot 1/Vo versus 1/[S].
5. Determine Km and Vmax using the Lineweaver-Burke Plot. Indicate on the plot how you determined Km and Vmax.
6. Assume there is one active site per enzyme molecule, what is the turnover
number of penicillinase under these experimental conditions?
7. How much time does it take for the penicillinase to hydrolyze one penicillin molecule
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