AbstractChiral alcohols are useful as intermediates for the synthesis of drugs. In the production of chiral alcohols, microbial enzymes are promising since high optical purity is required. Under these conditions, the reaction by resting cells is more convenient and inexpensive than by an enzyme reaction. Chiral 1,2‐propanediol and 2,3‐butanediol were obtained using cells expressing the enzyme which demonstrated high stereospecificity. By means of recombinant cells expressing the glycerol dehydrogenase of Hansenula polymorpha DL‐1, the medium was enriched with (S)‐1,2‐propanediol (98 % enantiometric excess, e.e.) during a 24‐h incubation, whereas the (R)‐form was removed from 100 mM of the racemate (R:S = 1:1). For an asymmetric reduction, a recombinant was constructed which also expressed the glucose dehydrogenase gene of Bacillus subtilis origin as an NADH reproducer. In the resting cell reaction, the pH control at 7.5 promoted the conversion from ketone to alcohol. (2R,3R)‐2,3‐butanediol (e.e. > 99.9 %; 308 mM) was produced from 800 mM acetoin (R:S = 3:4); (R)‐1,2‐propanediol (e.e. > 99.9 %; 550 mM) from 800 mM acetol in a 33‐h incubation by the addition of glucose and ketone with pH control. In this reaction, (S)‐forms of both 2,3‐butanediol and 1,2‐propanediol were not produced. The pH control and feeding of the substrates were uncomplicated. The production process of the chiral alcohol by the recombinants which expresses glycerol dehydrogenase proved convenient.