TABLE 1.

Kinetic data for the asymmetric amination of LA with EcGDH^K116C and EcGDH^K116Q/N348M.

Substrate	Enzyme	V max (U/mg)	K m (mM)	k cat (s−1)	k cat/K m (mM−1 s−1)
LA a	EcGDHK116C	0.109	1,340 ± 24.3	0.088 ± 0.0013	6.6 × 10–5
	EcGDHK116Q/N348M	2.82	824.0 ± 13.6	2.28 ± 0.19	2.77 × 10–3
NADPH b	EcGDHK116C	0.0834	0.0988 ± 0.003	0.067 ± 0.0037	0.68
	EcGDHK116Q/N348M	1.87	0.28 ± 0.07	1.51 ± 0.23	5.39

^aThe mixture composed of Tris-HCl (100 mM, pH 8.5), NH₄Cl-NH₄OH (0.8 M, pH 8.5), 0.2 mM NADPH, and different concentrations of LA (0–1,600 mM) was incubated at 30°C for 1 min before adding purified enzyme.

^bThe mixture containing Tris-HCl (100 mM, pH 8.5), NH₄Cl-NH₄OH (0.8 M, pH 8.5) different concentrations of NADPH (0–0.8 mM), and 80 mM LA was incubated at 30°C for 1 min before adding purified enzyme.