Table 1.

Kinetic parameters of CbADH and its mutants^a

ADH	Km [mM]	Kcat [min−1]	Kcat/Km [min−1 • mM−1]
CbADHb	7.2	8703	1209
TbADHb	5.8	2619	452
CbADH mutants
V224E-	7.1	7844	1105
S254K-	4.8	6069	1264
V224E/S254K-	6.2	5152	831
Q165E-	7.4	8144	1101
M304R-	6.3	7345	1166
Q165E/M304R-	7.1	15840	2231
Q165E/M304R/V224E/S254K	5.75	12542	2181
R238A/Q165E/M304R/V224E/S254K	7.47	15665	2097

^a Enzymatic activity was measured by following the formation of NADPH at 340 nm. Km values for 2-propanol were determined with varying concentrations of 2-propanol (0.5–100 mM), enzyme (5–120 nM), and 0.5 mM NADP in 100 mM Tris-HCl (pH 8.8) in a total volume of 1 ml in triplicate. Values are the averages of three experiments, and the individual measurements were within 10% of the quoted mean.

^b Values are from Bogin et al. (1998b).