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You have an enzyme that uses two substrates in a random reaction (the order of binding of the substrates is random).
You have an enzyme that uses two substrates in a random reaction (the order of binding of the substrates is random). And the reaction A + B à P is catalyzed by the enzyme bringtogetherase
You have carried out five sets of reaction rates Keeping B constant and varying A and then increasing B and Vary ing A again until all combinations of A and B are measured. In the reaction you used 0.02 mg of bringtogetherase which is a 120,000 Dalton protein.
The set of data is as follows
[A] mM Vo nmoles per min [B] mM
0.05 363.64 0.5
0.03333 322.58 0.5
0.0200 263.16 0.5
0.0125 208.33 0.5
0.01 181.81 0.5
0.0050 625 1
0.03333 555.56 1
0.0200 454.54 1
0.0125 357.14 1
0.01 312.5 1
0.05 952.38 2
0.03333 847.45 2
0.0200 714.29 2
0.0125 555.56 2
0.01 487.80 2
0.05 1250 5
0.03333 1111.11 5
0.0200 909.1 5
0.0125 724.64 5
0.01 632.91 5
0.05 1515.15 10
0.03333 1351.35 10
0.0200 1086.96 10
0.0125 847.46 10
0.01 740.74 10
Determine the Km for A, the Km for B the Vmax and the turnover number for the enzyme.
To get the Vmax you need to replot the intercepts [Vmax apparent] vs the other substrate.
