Abstract

Abstract The intracellular NADPH insufficient supply is the main bottleneck to the synthesis of chiral alcohols by asymmetric reduction with whole-cell catalysis. Herein, we provide a novel strategy to strengthen intracellular NADPH supply through introducing an NADP + -dependent glyceraldehyde 3-phosphate dehydrogenase ( gap B from Bacillus subtilis 168) into the Embden-Meyerhof pathway and a NAD kinase ( yfj B from E. coli MG1655) to further enhance the NADP(H) pool. A recombinant E. coli ( E. coli BL21 (DE3)/pETDuet-1- gap B- yue D&pET28a- yfj B) was constructed to co-express gap B and yfj B with a carbonyl reductase gene yue D together. The result showed that the intracellular NADPH amount increased by 134.4% with the strategy. To the model reaction (asymmetric reduction of acetophenone to S -phenyl ethanol), the yield was 3.7-fold with this strategy compared to the control. This provides a technological route for strengthening the intracellular NADPH supply in E. coli for biocatalysis and biosynthesis. Graphic Abstract