Table 1.

Kinetic parameters of NAD⁺-dependent oxidation reaction.

Enzymes	Substrates	Specific activitya (units/mg protein)	Km (mM)	kcat (min−1)	kcat/Km (min−1 mM−1)
Wild type	l-Alanine	0.0265 ± 0.0015	18.2 ± 5.0	2.64 ± 0.55	0.147 ± 0.012
	l-2-Aminobutyrate	0.136 ± 0.010	6.00 ± 0.29	6.71 ± 0.16	1.12 ± 0.03
	l-2-Aminovalerate	0.396 ± 0.030	0.306 ± 0.009	6.56 ± 0.08	21.5 ± 0.4
	l-Leucine	0.248 ± 0.010	1.32 ± 0.10	10.5 ± 0.4	8.00 ± 0.28
	N-Methyl-l-alanine	0.526 ± 0.025	0.406 ± 0.044	11.6 ± 0.7	28.6 ± 2.1
	l-Proline	0.384 ± 0.015 (0.0376 ± 0.0005)b	1.12 ± 0.12 (13.1 ± 1.2)b	13.8 ± 1.0 (3.27 ± 0.24)b	12.3 ± 0.4 (0.250 ± 0.006)b
	l-Pipecolate	0.219 ± 0.020	0.608 ± 0.115	12.2 ± 1.3	20.3 ± 1.8
V224D/A228K	l-Alanine	0.0133 ± 0.0010	36.9 ± 0.9	2.12 ± 0.02	0.0575 ± 0.0008
	l-2-Aminovalerate	0.666 ± 0.050	2.13 ± 0.30	7.72 ± 0.82	3.64 ± 0.12
	l-Proline	1.51 ± 0.10	0.444 ± 0.008	36.5 ± 0.9	82.2 ± 1.1

^aUnder standard assay conditions in “Experimental procedures”. All substrate concentrations were 10 mM.

^bNADP⁺-dependent activity.