초록
(S)-N-Boc-3-hydroxypiperidine ((S)-NBHP) is an important chiral intermediate for the synthesis of ibrutinib, an anticancer drug targeting B-cell malignancies. An NADPH-dependent reductase (YDR541C) from Saccharomyces cerevisiae was isolated and found to show excellent catalytic activity in the production of (S)-NBHP. The reductase YDR541C was cloned and overexpressed in Escherichia coli BL21 (DE3), purified to homogeneity, and its catalytic properties were studied. Furthermore, an ethyl caprylate-water (1:1, v/v) biphasic system was introduced to alleviate product inhibition. After optimization of the reaction, as much as 1200mM N-Boc-piperidin-3-one (NBPO) (240g/L) was asymmetrically reduced to (S)-NBHP within 6h, resulting in a yield of 99%, an enantioselectivity of >99.5% ee, and a total turnover number (TTN) of 8000. These results indicate great potential for industrial application.