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  • Enhanced 2,3-butanediol production in recombinant Klebsiella pneumoniae via overexpression of synthesis-related genes.

Enhanced 2,3-butanediol production in recombinant Klebsiella pneumoniae via overexpression of synthesis-related genes.

Journal of microbiology and biotechnology (2012-07-21)
Borim Kim, Soojin Lee, Joohong Park, Mingshou Lu, Minkyu Oh, Youngrok Kim, Jinwon Lee
ABSTRACT

2,3-Butanediol (2,3-BD) is a major metabolite produced by Klebsiella pneumoniae KCTC2242, which is a important chemical with wide applications. Three genes important for 2,3-BD biosynthesis acetolactate decarboxylase (budA), acetolactate synthase (budB), and alcohol dehydrogenase (budC) were identified in K. pneumoniae genomic DNA. With the goal of enhancing 2,3-BD production, these genes were cloned into pUC18K expression vectors containing the lacZ promoter and the kanamycin resistance gene to generate plasmids pSB1-7. The plasmids were then introduced into K. pneumoniae using electroporation. All strains were incubated in flask experiments and 2,3-BD production was increased by 60% in recombinant bacteria harboring pSB04 (budA and budB genes), compared with the parental strain K. pneumoniae KCTC2242. The maximum 2,3-BD production level achieved through fedbatch fermentation with K. pneumoniae SGJSB04 was 101.53 g/l over 40 h with a productivity of 2.54 g/l.h. These results suggest that overexpression of 2,3-BD synthesisrelated genes can enhance 2,3-BD production in K. pneumoniae by fermentation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(2S,3S)-(+)-2,3-Butanediol, 97%
Sigma-Aldrich
meso-2,3-Butanediol, 99%
Sigma-Aldrich
(2R,3R)-(−)-2,3-Butanediol, 97%
Sigma-Aldrich
2,3-Butanediol, 98%