FAD-dependent D-lactate dehydrogenase catalyzes the dehydrogenation of D-lactate to pyruvate,which is a key enzyme for metabolism of D-lactic acid.The coding gene dld of an FDA-dependent D-lactate dehydrogenase was cloned from the genome of Escherichia coli B0013 and expressed in E.coli JM109.The temperature and pH optima of the purified recombinant D-lactate dehydrogenase were 35 ℃ and 7.0,respectively,while good stability in 30-40 ℃ and pH 5.0-7.5 were revealed.The enzyme was significantly activated by Zn2+,while strongly inhibited by Mn2+,Cu2+,Mg2+ and SDS.It was highly specific for D-lactic acid,with Km of 0.057 mg/mL,Vmax of 0.054 μmol/(mL·min),kcat of 0.275 min-1,and kcat/Kmof 0.435 mL/(mg·min) at 35 ℃ and pH 7.0.The enzyme is potentially applicable in quantitative detection of D-lactic acid.
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