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Mechanism of 2,3-butanediol stereoisomer formation in Klebsiella pneumoniae

  • Applied microbial and cell physiology
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Abstract

Klebsiella pneumoniae is known to produce meso-2,3-butanediol and 2S,3S-butanediol, whereas 2R,3R-butanediol was detected in the culture broth of K. pneumoniae CGMCC 1.6366. The ratio of 2R,3R-butanediol to all isomers obtained using glycerol as the carbon source was higher than that obtained using glucose as the carbon source. Therefore, enzymes involved in glycerol metabolism are likely related to 2R,3R-butanediol formation. In vitro reactions show that glycerol dehydrogenase catalyzes the stereospecific conversion of R-acetoin to 2R,3R-butanediol and S-acetoin to meso-2,3-butanediol. Butanediol dehydrogenase exhibits high (S)-enantioselectivity in ketone reduction. Genes encoding glycerol dehydrogenase, α-acetolactate decarboxylase, and butanediol dehydrogenase were individually disrupted in K. pneumoniae CGMCC 1.6366, and the 2,3-butanediol synthesis characteristics of these mutants were investigated. K. pneumoniae ΔdhaD lost the ability to synthesize 2R,3R-butanediol. K. pneumoniae ΔbudA showed reduced 2R,3R-butanediol synthesis. However, K. pneumoniae ΔbudC produced a high level of 2R,3R-butanediol, and R-acetoin was accumulated in the broth. The metabolic characteristics of these mutants and in vitro experiment results demonstrated the mechanism of the 2,3-butanediol stereoisomer synthesis pathway. Glycerol dehydrogenase, encoded by dhaD, exhibited 2R,3R-butanediol dehydrogenase activity and was responsible for 2R,3R-butanediol synthesis from R-acetoin. This enzyme also contributed to meso-2,3-butanediol synthesis from S-acetoin. Butanediol dehydrogenase, encoded by budC, was the only enzyme that catalyzed the conversion of diacetyl to S-acetoin and further to 2S,3S-butanediol.

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Acknowledgments

This work was financially supported by the National Natural Science Foundation of China (grant no. 20906076).

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Authors and Affiliations

  1. Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science & Technology), Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People’s Republic of China

    Chuan Chen & Min Wang

  2. Lab of Biorefinery, Shanghai Advanced Research Institute, Chinese Academy of Sciences, No. 99 Haike Road, Pudong, Shanghai, 201210, People’s Republic of China

    Chuan Chen, Dong Wei, Jiping Shi & Jian Hao

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  1. Chuan Chen
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  2. Dong Wei
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  3. Jiping Shi
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  4. Min Wang
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Correspondence to Min Wang or Jian Hao.

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Chen, C., Wei, D., Shi, J. et al. Mechanism of 2,3-butanediol stereoisomer formation in Klebsiella pneumoniae . Appl Microbiol Biotechnol 98, 4603–4613 (2014). https://doi.org/10.1007/s00253-014-5526-9

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  • DOI: https://doi.org/10.1007/s00253-014-5526-9

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