Characterization of d-succinylase from Cupriavidus sp. P4-10-C and its application in d-amino acid synthesis

d -Amino acids are important building blocks for various compounds, such as pharmaceuticals and agrochemicals. A more cost-effective enzymatic method for d -amino acid production is needed in the industry. We improved a one-pot enzymatic method for d -amino acid production by the dynamic kinetic resolution of N-succinyl amino acids using two enzymes: d -succinylase (DSA) from Cupriavidus sp. P4-10-C, which hydrolyzes N-succinyl- d -amino acids enantioselectively to their corresponding d -amino acid, and N-succinyl amino acid racemase (NSAR, EC.4.2.1.113) from Geobacillus stearothermophilus NCA1503. In this study, DSA and NSAR were purified and their properties were investigated. The optimum temperature of DSA was 50°C and it was stable up to 55°C. The optimum pH of DSA and NSAR was around 7.5. In d -phenylalanine production, the optical purity of product was improved to 91.6% ee from the examination about enzyme concentration. Moreover, 100 mM N-succinyl- dl -tryptophan was converted to d -tryptophan at 81.8% yield with 94.7% ee. This enzymatic method could be useful for the industrial production of various d -amino acids.