Recombinant plasmids containing D-arabino-1, 4-lactone oxidase gene, transformants, and methods for mass production of D-erythroascholic acid and L-ascholic acid using the same

摘要

The products of two parts of D-arabino-1,4-lactone oxidase involved in D-erythroaskolbinic acid biosynthesis in Saccharomyces cerevisiae synthesized by gene amplification using primers were introduced into pGEM-T Easy vector. Intact D-arabino-1,4-lactone oxidase was obtained through Sac I, a restriction enzyme within. They were overexpressed by the expression vector pET-3a from E. coli, the most well-known expression system of prokaryotes. Among them, remarkable enzymatic activity was observed in the water-soluble portion of D-arabino-1,4-lactone oxidase. It can be used to produce excess (or large quantities) of D-erythroascorbic acid and L-ascorbic acid.