Four strains of yeast with reduction activity of chloroacetophenones were screened, in which Saccharomyces cerevisiae B5 showed best reduction activity and stereoselectivity. High optical purity (R)-2'-chloro-1-phenylethanol can be obtained with Saccharomyces cerevisiae B5 as biocatalyst. The influence of several co-substrates on the enantiometric excess (ee%) and yield of (R)-2'-chloro-1-phenylethanol was evaluated. 5% (v/v) ethanol is optimal cosubstrate for (R)-2'-chloro-1-phenylethanol formation. The optimal bioconversion conditions of 2'-chloroacetophenone catalyzed by Saccharomyces cerevisiae B5 are as follows: pH 8.0, 25 oC and 24 h. The yield and the enantiometric excess of (R)-2'-chloro-1-phenylethanol can both reach more than 99% with 10.75 g/l Saccharomyces cerevisiae B5 (the cell dry weight) and 1 g/l 2'-chloroacetophenone used in the biotransformation.