(2S,3S)-2,3-Butanediol ((2S,3S)-2,3-BD) is a crucial chiral compound that acts as an excellent building block in asymmetric synthesis of highly valuable chiral compounds. However, the low concentration and optical purity of (2S,3S)-2,3-BD produced in previous studies limited its applications. In the present work, the gene encoding 2,3-butanediol dehydrogenase from an Enterobacter cloacae ssp. dissolvens strain SDM was cloned and expressed in Escherichia coli. Whole cells of the recombinant E. coli was used to produce (2S,3S)-2,3-BD from diacetyl. Under optimal conditions, high-optical-purity (2S,3S)-2,3-BD (purity >99%) was obtained with concentrations of 16.1 g l(-1) and 26.8 g l(-1) in batch and fed-batch conversions, respectively. Thus, the process might be a promising alternative for the production of (2S,3S)-2,3-BD. (C) 2011 Elsevier Ltd. All rights reserved.