Table 3. Kinetic Parameters for Wild-Type and Mutant EIZS Enzymes.
enzyme | kcat (ms–1) | KM (nM) | kcat/KM (M–1 s–1) |
---|---|---|---|
WT | 84 ± 1 | 330 ± 20 | 260000 ± 10000 |
Aromatic Mutants | |||
F95A | 1.8 ± 0.2 | 672 ± 2 | 2700 ± 300 |
F95V | 7.4 ± 0.4 | 1700 ± 300 | 4400 ± 600 |
F95M | 3.16 ± 0.09 | 275 ± 4 | 11500 ± 500 |
F95H | 1.72 ± 0.02 | 650 ± 30 | 2600 ± 100 |
F96Aa | 0.24 ± 0.02 | 800 ± 100 | 310 ± 60 |
F96V | 1.15 ± 0.02 | 580 ± 30 | 199 ± 7 |
F96L | 2.57 ± 0.02 | 820 ± 60 | 3100 ± 200 |
F96Y | 0.19 ± 0.01 | 1800 ± 100 | 107 ± 2 |
F96W | 0.35 ± 0.02 | 310 ± 70 | 1200 ± 400 |
F198Aa | 0.30 ± 0.02 | 1200 ± 200 | 250 ± 50 |
F198V | 1.66 ± 0.09 | 2800 ± 400 | 600 ± 60 |
F198L | 4.75 ± 0.05 | 4200 ± 400 | 1100 ± 100 |
F198Y | 15.04 ± 0.06 | 4100 ± 100 | 3600 ± 100 |
W203Fa | 0.34 ± 0.03 | 1500 ± 200 | 230 ± 40 |
W325F | 22 ± 2 | 160 ± 60 | 14000 ± 5000 |
F332A | 0.105 ± 0.008 | 360 ± 90 | 290 ± 80 |
Aliphatic Mutants | |||
L72A | 8.81 ± 0.09 | 3000 ± 100 | 2900 ± 100 |
L72V | 44 ± 3 | 600 ± 100 | 70000 ± 20000 |
L72M | 11 ± 2 | 3200 ± 900 | 3400 ± 400 |
A236G | 66 ± 2 | 450 ± 50 | 150000 ± 20000 |
A236M | – | – | – |
A236F | 0.0424 ± 0.0007 | 230 ± 50 | 190 ± 40 |
V329A | 126 ± 7 | 470 ± 80 | 270000 ± 50000 |
V329G | 19.8 ± 0.1 | 1590 ± 60 | 12500 ± 500 |
V329I | 22.8 ± 0.3 | 770 ± 40 | 29000 ± 1000 |
V329M | 0.640 ± 0.004 | 6500 ± 300 | 98 ± 3 |
Mutants previously reported (ref (13)).