Selective Production of 2,3-Butanediol and Acetoin by a Newly Isolated Bacterium Klebsiella oxytoca M1

Abstract

A newly isolated bacterium, designated as Klebsiella oxytoca M1, produced 2,3-butanediol (2,3-BDO) or acetoin selectively as a major product depending on temperature in a defined medium. K. oxytoca M1 produced 2,3-BDO mainly (0.32 similar to 0.34 g/g glucose) at 30 A degrees C while acetoin was a major product (0.32 similar to 0.38 g/g glucose) at 37 A degrees C. To investigate factors affecting product profiles according to temperature, the expression level of acetoin reductase (AR) that catalyzes the conversion of acetoin to 2,3-BDO was analyzed using crude protein extracted from K. oxytoca M1 grown at 30 and 37 A degrees C. The AR expression at 37 A degrees C was 12.8-fold lower than that at 30 A degrees C at the stationary phase and reverse transcription PCR (RT-PCR) analysis of the budC (encoding AR) was also in agreement with the AR expression results. When AR was overexpressed using K. oxytoca M1 harboring pUC18CM-budC, 2,3-BDO became a major product at 37 A degrees C, indicating that the AR expression level was a key factor determining the major product of K. oxytoca M1 at 37 A degrees C. The results in this study demonstrate the feasibility of using K. oxytoca M1 for the production of not only 2,3-BDO but also acetoin as a major product.