The embodiment of the application belongs to the technical field of
malate dehydrogenase preparation, and relates to a
thermus thermophilus malate dehydrogenase recombinant
gene and a preparation method and application thereof, comprising the following steps: adopting a
gene sequence of
thermus thermophilus malate dehydrogenase, optimizing the
gene sequence of the
thermus thermophilus malate
dehydrogenase by using the synonymous codon preference of
escherichia coli, obtaining a thermus thermophilus malate
dehydrogenase recombinant gene, constructing a thermus thermophilus malate
dehydrogenase recombinant
plasmid based on the thermus thermophilus malate dehydrogenase recombinant gene, transforming the thermus thermophilus malate dehydrogenase recombinant
plasmid into
escherichia coli and culturing, obtaining a
bacteria liquid for expressing malate dehydrogenase, purifying the
bacteria liquid for expressing malate dehydrogenase, and obtaining malate dehydrogenase. The malate dehydrogenase expressed by the application has high yield, short
production cycle and
high activity.