Lactobacillus leichimanii D-Lactate Dehydrogenase 클로닝과 분리정제 및 Lactate Chirality 정성 정량 분석

Cloning and Purification of Recombinant Lactobacillus leichimanii D-Lactate Dehydrogenase and its Use in the Quantification of Lactate Chirality

The D-lactate level in body fluids is an important factor associated with various metabolic diseases. Nevertheless, it is difficult to quantify lactate isomers in clinical laboratories. Here, a simple enzymatic assay employing recombinant D-lactate dehydrogenase to assess the level of D(-)-lactate in plasma or serum has been described. The D-lactate dehydrogenase gene from Lactobacillus leichmannii was cloned and expressed using E. coli expression system. The purified enzyme showed enantioselective activity, and D-lactate concentration was measured by estimating the level of NADH, a by-product of the enzymatic reaction. This rapid and accurate enzymatic assay can be applied to evaluate patients suffering from metabolite-related diseases.