Table 3.

Kinetic Parameters of the Wild-Type and Mutant Enzymes

Enzyme	Km (mM)	Vmax (U/mg)	kcat (s−1)	kcat/Km (M−1 s−1)
TmBglA	0.38 ± 0.02	526.42 ± 9.42	452.27 ± 8.25	1.21 ± 0.14 × 106
N222F	0.20 ± 0.01	713.54 ± 13.57	613.43 ± 10.39	3.00 ± 0.50 × 106
N223C	0.40 ± 0.01	753.43 ± 12.03	647.16 ± 9.74	1.61 ± 0.06 × 106
N223Q	0.56 ± 0.04	895.67 ± 14.23	769.72 ± 12.14	1.40 ± 0.24 × 106
G224A	0.35 ± 0.03	718.34 ± 8.58	617.32 ± 6.75	1.77 ± 0.06 × 106
Y295F	0.61 ± 0.09	6.16 ± 0.23	5.29 ± 0.14	8.74 ± 0.51 × 103
F414S	2.29 ± 0.01	7.49 ± 0.01	6.42 ± 0.01	2.81 ± 0.01 × 103

Note. The kinetic parameters were determined at their optimal pH and temperature for the substrate concentrations ranging from 0.2 to 2.0 mM for pNPG using the standard assay as described in “Materials and Methods.”