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Identification and characterization of a short-chain acyl dehydrogenase from Klebsiella pneumoniae and its application for high-level production of l-2,3-butanediol

  • Genetics and Molecular Biology of Industrial Organisms
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Klebsiella pneumoniae synthesize large amounts of l-2,3-butanediol (l-2,3-BD), but the underlying mechanism has been unknown. In this study, we provide the first identification and characterization of an l-2,3-BD dehydrogenase from K. pneumoniae, demonstrating its reductive activities toward diacetyl and acetoin, and oxidative activity toward l-2,3-BD. Optimum pH, temperature, and kinetics determined for reductive and oxidative reactions support the preferential production of 2,3-BD during cell growth. Synthesis of l-2,3-BD was remarkably enhanced by increasing gene dosage, reaching levels that, to the best of our knowledge, are the highest achieved to date.

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References

  1. Celińska E, Grajek W (2009) Biotechnological production of 2,3-butanediol-current state and prospects. Biotechnol Adv 27:715–725. doi:10.1016/j.biotechadv.2009.05.002

    Article  PubMed  Google Scholar 

  2. Cherepanov PP, Wackernagel W (1995) Gene disruption in Escherichia coli: TcR and KmR cassettes with the option of Flp-catalyzed excision of the antibiotic-resistance determinant. Gene 158:9–14. doi:10.1016/0378-1119(95)00193-A

    Article  CAS  PubMed  Google Scholar 

  3. Datsenko KA, Wanner BL (2000) One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97:6640–6645. doi:10.1073/pnas.120163297

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Fournet-Fayard S, Joly B, Forestier C (1995) Transformation of wild type Klebsiella pneumoniae with plasmid DNA by electroporation. J Microbiol Methods 24:49–54. doi:10.1016/0167-7012(95)00053-4

    Article  Google Scholar 

  5. Li L, Wang Y, Zhang L, Ma C, Wang A, Tao F, Xu P (2011) Biocatalytic production of (2S,3S)-2,3-butanediol from diacetyl using whole cells of engineered Escherichia coli. Bioresour Technol 115:111–116. doi:10.1016/j.biortech.2011.08.097

    Article  PubMed  Google Scholar 

  6. Luo LH, Seo JW, Baek JO, Oh BR, Heo SY, Hong WK, Kim DH, Kin CH (2011) Identificaiton and characterization of the propanediol utilization protein PduP of Lactobacillus reuteri for 3-hydroxypropionic acid production from glycerol. Appl Microbiol Biotechnol 89:697–703. doi:10.1007/s00253-010-2887-6

    Article  CAS  PubMed  Google Scholar 

  7. Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29:351–364. doi:10.1016/j.biotechadv.2011.01.007

    Article  CAS  PubMed  Google Scholar 

  8. Jurchescu IM, Hamann J, Zhou X, Ortmann T, Kuenz A, Pruße U, Lang S (2013) Enhanced 2,3-butanediol production in fed-batch cultures of free and immobilized Bacillus licheniformis DSM 8785. Appl Microbiol Biotechnol 97:6715–6723. doi:10.1007/s00253-013-4981-z

    Article  CAS  PubMed  Google Scholar 

  9. Ma C, Wang A, Qin J, Li L, Ai X, Jiang T, Tang H, Xu P (2009) Enhanced 2,3-butanediol production by Klebsiella pneumoniae SDM. Appl Microbiol Biotechnol 82:49–57. doi:10.1007/s00253-008-1732-7

    Article  CAS  PubMed  Google Scholar 

  10. Oh BR, Seo JW, Heo SY, Hong WK, Luo LH, Joe MH, Park DH, Kim CH (2011) Efficient production of ethanol from crude glycerol by a Klebsiella pneumoniae mutant strain. Bioresour Technol 102:3918–3922. doi:10.1016/j.biortech.2010.12.007

    Article  CAS  PubMed  Google Scholar 

  11. Oh BR, Seo JW, Heo SY, Hong WK, Luo LH, Kim SH, Park DH, Kim CH (2012) Optimization of culture conditions for 1,3-propanediol production from glycerol using a mutant strain of Klebsiella pneumoniae. Appl Biochem Biotechnol 166:127–137. doi:10.1007/s12010-011-9409-6

    Article  CAS  PubMed  Google Scholar 

  12. Oh BR, Hong WK, Heo SY, Joe MH, Seo JW, Kim CH (2013) The role of aldehyde/alcohol dehydrogenase (AdhE) in ethanol production from glycerol by Klebsiella pneumoniae. J Ind Microbiol Biotechnol 40:227–233. doi:10.1007/s10295-012-1224-8

    Article  CAS  PubMed  Google Scholar 

  13. Oh BR, Heo SY, Lee SM, Hong WK, Park JM, Jung YR, Kim DH, Sohn JH, Seo JW, Kim CH (2014) Production of isobutanol from crude glycerol by a genetically-engineered Klebsiella pneumoniae strain. Biotechnol Lett 36:397–402. doi:10.1007/s10529-013-1383-3

    Article  CAS  Google Scholar 

  14. Oliver JW, Machado IM, Yoneda H, Atsumi S (2013) Cyanobacterial conversion of carbon dioxide to 2,3-butanediol. Proc Natl Acad Sci 110:1249–1254. doi:10.1073/pnas.1213024110

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  15. Park JM, Oh BR, Seo JW, Hong WK, Yu A, Sohn JH, Kim CH (2013) Efficient production of ethanol from empty palm fruit bunch fibers by fed-batch simultaneous saccharification and fermentation using Saccharomyces cerevisiae. Appl Biochem Biotechnol 170:1807–1814. doi:10.1007/s12010-013-0314-z

    Article  CAS  PubMed  Google Scholar 

  16. Syu MJ (2001) Biological production of 2,3-butanediol. Appl Microbiol Biotechnol 55:10–18. doi:10.1007/s002530000486

    Article  CAS  PubMed  Google Scholar 

  17. Ui S, Matsuyama N, Masuda H, Muraki H (1984) Mechanism for the formation of 2,3-butanediol stereoisomers in Klebsiella pneumoniae. J Ferment Technol 62:551–559. doi:10.1007/s00253-014-5526-9

    CAS  Google Scholar 

  18. Ui S, Takusagawa Y, Ohtsuki T, Mimura A, Ohkuma M, Kudo T (2001) Stereochemical applications of the expression of the l-2,3-butanediol dehydrogenase gene in Escherichia coli. Lett Appl Microbiol 32:93–98. doi:10.1046/j.1472-765x.2001.00869

    Article  CAS  PubMed  Google Scholar 

  19. Wang Z, Song Q, Yu M, Wang Y, Xiong B, Zhang Y, Zheng J, Ying X (2014) Characterization of a stereospecific acetoin (diacetyl) reductase from Rhodococcus erythropolis WZ010 and its application for the synthesis of (2S,3S)-2,3-butanediol. Appl Microbiol Biotechnol 98:641–650. doi:10.1007/s00253-013-4870-5

    Article  CAS  PubMed  Google Scholar 

  20. Yan YJ, Lee CC, Liao JC (2009) Enantioselective synthesis of pure (R, R)-2,3-butanediol in Escherchia coli with stereospecific secondary alcohol dehydrogenase. Org Biomol Chem 7:3914–3917. doi:10.1039/b913501d

    Article  CAS  PubMed  Google Scholar 

  21. Zhang GL, Wang CW, Li C (2012) Cloning, expression and characterization of meso-2,3-butanediol dehydrogenase from Klebsiella pneumoniae. Biotechnol Lett 34:1519–1523. doi:10.1007/s10529-012-0933-4

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the Korea Research Foundation (KRF) for Basic Research Promotion funded by the Korea Government (MEST) and the Joint Degree and Research Center funded by the Korea Research Council of Fundamental Science & Technology.

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Authors and Affiliations

  1. Biorefinery Research Center, Jeonbuk Branch Institute, KRIBB, Jeongeup, Jeonbuk, 580-185, South Korea

    Jang Min Park, Won-Kyung Hong, Sung-Mok Lee, Sun-Yeon Heo, You Ree Jung, In Yeong Kang, Baek-Rock Oh, Jeong-Woo Seo & Chul Ho Kim

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  1. Jang Min Park
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  2. Won-Kyung Hong
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Correspondence to Jeong-Woo Seo.

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Park, J.M., Hong, WK., Lee, SM. et al. Identification and characterization of a short-chain acyl dehydrogenase from Klebsiella pneumoniae and its application for high-level production of l-2,3-butanediol. J Ind Microbiol Biotechnol 41, 1425–1433 (2014). https://doi.org/10.1007/s10295-014-1483-7

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  • DOI: https://doi.org/10.1007/s10295-014-1483-7

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