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The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli by Employing New CoA Transferases

재조합 대장균에서 새로운 코엔자임 에이 트랜스퍼레이즈를 이용한 젖산을 모노머로 함유한 폴리하이드록시알칸산 생산 연구

  • Kim, You Jin (Department of Environmental Engineering and Energy, Myongji University) ;
  • Chae, Cheol Gi (Department of Environmental Engineering and Energy, Myongji University) ;
  • Kang, Kyoung Hee (Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology) ;
  • Oh, Young Hoon (Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology) ;
  • Joo, Jeong Chan (Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology) ;
  • Song, Bong Keun (Center for Bio-based Chemistry, Division of Convergence Chemistry, Korea Research Institute of Chemical Technology) ;
  • Lee, Sang Yup (Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical and Biomolecular Engineering (BK21 Program), Center for Systems and Synthetic Biotechnology, and Institute for the BioCentury, KAIST) ;
  • Park, Si Jae (Department of Environmental Engineering and Energy, Myongji University)
  • 김유진 (명지대학교 환경에너지공학과) ;
  • 채철기 (명지대학교 환경에너지공학과) ;
  • 강경희 (한국화학연구원 바이오화학연구센터) ;
  • 오영훈 (한국화학연구원 바이오화학연구센터) ;
  • 주정찬 (한국화학연구원 바이오화학연구센터) ;
  • 송봉근 (한국화학연구원 바이오화학연구센터) ;
  • 이상엽 (한국과학기술원 생명화학공학과) ;
  • 박시재 (명지대학교 환경에너지공학과)
  • Received : 2016.01.03
  • Accepted : 2016.01.29
  • Published : 2016.03.31
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Abstract

Several CoA transferases from Clostridium beijerinckii, C. perfringens and Klebsiella pneumoniae were examined for biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) in recombinant Escherichia coli XL1-Blue strain. The CB3819 gene and the CB4543 gene from C. beijerinckii, the pct gene from C. perfringens and the pct gene from K. pneumoniae, which encodes putative CoA transferase gene, respectively, was co-expressed with the Pseudomonas sp. MBEL 6-19 phaC1437 gene encoding engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 ($PhaC1_{Ps6-19}$) to examine its activity for the construction of key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli XL1-Blue expressing the phaC1437 gene and CB3819 gene synthesized poly(3-hydroxybutyrate) [P(3HB)] homopolymer to the P(3HB) content of 60.5 wt% when it was cultured in a chemically defined medium containing 20 g/L of glucose and 2 g/L of sodium 3-hydroxybutyrate. Expression of the phaC1437 gene and CB4543 gene in recombinant E. coli XL1-Blue also produced P(3HB) homopolymer to the P(3HB) content of 51.2 wt% in the same culture condition. Expression of the phaC1437 gene and the K. pneumoniae pct gene in recombinant E. coli XL1-Blue could not result in the production of PHAs in the same culture condition. However, the recombinant E. coli XL1-Blue expressing the phaC1437 gene and the C. perfringens gene could produce poly(3-hydroxybutyrate-co-lactate [P(86.4mol%3HB-co-13.7 mol%LA) up to the PHA content of 10.6 wt% in the same culture condition. Newly examined CoA transfereases in this study may be useful for the construction of engineered E. coli strains to produce PHA containing novel monomer such lactate.

Keywords

References

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  1. Biosynthesis of 2-Hydroxyacid-Containing Polyhydroxyalkanoates by Employing butyryl-CoA Transferases in Metabolically Engineered Escherichia coli 2017, https://doi.org/10.1002/biot.201700116
  2. A Review of the Recent Developments in the Bioproduction of Polylactic Acid and Its Precursors Optically Pure Lactic Acids vol.26, pp.21, 2016, https://doi.org/10.3390/molecules26216446