Hydrogenation of pyruvic acid by hydrogen with the combination of hydrogenase and lactate dehydrogenase

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The regeneration of nicotinamide-adenine dinucleotide (reduced form NADH) by enzymatic systems is of great advantage in producing useful compounds for the simplification of certain systems, as reported previously [l-6]. In this study, hydrogenase from Akal~ms eutrophus was used for the regeneration of NADH from NAD+ with hydrogen gas as a reducing agent; hydrogenation of pyruvic acid was attempted by a combination of this system with lactate dehydrogenase, as shown in Scheme 1. The hydrogenase from Alcaligenes eutrophus was partly purified according to the literature [ 11. The activity (1 unit) of hydrogenase was to reduce 1 pmol of NAD’ for 1 min in the system containing hydrogenase and NADc (5.0X lop4 mol drn3) in 4.0 ml of a 5.0X 10e3 mol dmP3 phosphate buffer (pH 7.0) under 300 torr hydrogen pressure at 30 “C. Lactate dehydrogenase (LDH) from rabbit muscles was obtained from Sigma Co.

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