Abstract: R-Ethyl-4-chloro-3-hydroxybutanoate（R-CHBE）is used as a chiral building block for the synthesis of pharmaceutical target compounds. R-CHBE can be produced by the asymmetric reduction of ethyl 4-chloro-3-oxobutanoate（COBE）. However，effective cofactor regeneration is a key step for the application of most oxidoreductases. In this research，the NADPH-dependent aldehyde reductase gene（alr）was cloned from the genome of Sporobolomyces salmonicolor ZJU010，and the recombinant E.coli BL21/pET-alr strain was constructed. Asymmetric reduction of COBE to CHBE was performed using the recombinant E.coli BL21/pET-alr coupling with E.coli BL21/pET-gdh. The highest reduction activity was obtained by recombinant resting cells at 6∶1 weight ratio of E.coli BL21/pET-alr with E.coli BL21/pET-gdh. Yield and the enantiomeric excess（ee）of R-CHBE reached to 95.88% and 96%，respectively，after 22h bioconversion without adding coenzyme NADPH or NADP+. R-CHBE could be effectly produced using the recombinant E.coli BL21/pET-alr coupling with E.coli BL21/pET-gdh for NADPH regenerantion．