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http://dx.doi.org/10.7841/ksbbj.2016.31.1.27

Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli by Employing New CoA Transferases  

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)
Publication Information
KSBB Journal / v.31, no.1, 2016 , pp. 27-32 More about this Journal
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
Escherichia coli; Polyhydroxyalkanoate (PHA); Lactate-containing PHA; Propionyl-CoA transferase;
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