Altering coenzyme specificity of Pichia stipitis xylose reductase by the semi-rational approach CASTing

Table 4 Comparison of the degree of alteration of coenzyme specificity for various mutants.

Enzyme	K_m[mM]		K_m^NADPH/K_m^NADH	k_cat [Min^-1]		k_cat^NADH/k_cat^NADPH	(k_cat/K_m^NADH)/(k_cat/K_m^NADPH)
	NADH	NADPH		NADH	NADPH
Wild-type NT-XR	0.0106	0.0062	0.58	923	1650	0.56	0.31
2-2C12	0.147	0.472	2.90	720	158	4.56	13.2
NT-K270R/N272D	0.058	0.142	2.45	1075	955	1.13	2.93
Wild-type^[a]	0.021	0.009	0.43	NA^[d]	NA^[d]	NA^[d]	NA^[d]
Cys130Ser^[a]	0.021	0.032	1.52	NA^[d]	NA^[d]	NA^[d]	NA^[d]
Wild-type^[b]	0.028	0.026	0.93	NA^[d]	NA^[d]	NA^[d]	NA^[d]
Lys270Met^[b]	0.031	0.131	4.30	NA^[d]	NA^[d]	NA^[d]	NA^[d]
Wild-type^[c]	0.0305	0.0025	0.082	415	630	0.66	0.05
R276H^[c]	0.017	0.0017	0.1	408	16	25.5	2.62
K270R/N272D^[c]	0.138	2.81	20.4	706	1850	0.65	7.29

[a] Data were quoted from Y. Y. Zhang et al [2]. [b] Data were quoted from M. Kostrzynska et al, and the substrate used in the reaction mixture was DL-glyceraldehyde instead of xylose [3]. [c] Data were quoted from S. Watanabe et al [21]. [d] NA, not available.

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