PURPOSE: To efficiently produce a useful substance in high enzyme activity even at an alkali side in the presence of a dehydrogenase by using a culture mixture of NADH oxidase producing bacterium having optimum growth pH in an alkali range.
CONSTITUTION: A bacterium such as Escherichia coli K-12 base transformed with a plasmid having at least a DNA range containing a gene of managing biosynthesis of alkali-philic, anaerobic DADN oxidase is cultured by using an ordinarily useful medium under an aerobic condition. Then, the cultured cells are ground by ultrasonic treatment, etc., the prepared cell-free extracted solution is separated and purified by salting-out, etc., so that NADH oxidase can be collected. Then a culture mixture of the above-mentioned transformed bacterium or a treated material containing NADS is conjugated with a dehydrogenase such as malate dehydrogenase having NAD+ as a coenzyme so that the resulting substance can be used for dehydrogenation reaction of substrate and is utilized for production of various useful substances and biochemical reactions.
YAMAGATA HISASHI
KURUSU YASUROU
TERASAWA MASATO
YUGAWA HIDEAKI
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