2,3-Butanediol is an attractive product for biorefinery due to its various applications and industrial potential. Enterobacter aerogenes is also considered as a major 2,3-butanediol producer. This study is intended to enhance the production of 2,3-butanediol with addition of acetate into the culture media using E. aerogenes ATCC 29007 and to determine the mechanism of improved 2,3-butanediol production. 2,3-Butanediol production was increased by 34.2% with the addition of acetate in batch culture compared to that without acetate in 130 of glucose. This increase was caused by buffering effects and upregulation of the transcription levels of genes involved in 2,3-butanediol production through acetate addition. (C) 2016 Elsevier Ltd. All rights reserved.