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KMID : 0380020140290060443
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Korean Journal of Biotechnology and Bioengineering
2014 Volume.29 No. 6 p.443 ~ p.447
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Biosynthesis of Lactate-containing Polyhydroxyalkanoates in Recombinant Escherichia coli from Sucrose
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Oh Young-Hoon
Kang Kyoung-Hee
Shin Ji-Hoon
Song Bong-Keun
Lee Seung-Hwan
Lee Sang-Yup
Park Si-Jae
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Abstract
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Biosynthesis of lactate-containing polyhydroxyalkanoates (PHAs) was examined in recombinant Escherichia coli W strain from sucrose. The Pseudomonas sp. MBEL 6-19 phaC1437 gene and the Clostridium propionicum pct540 gene, which encode engineered Pseudomonas sp. MBEL 6-19 PHA synthase 1 (PhaC1Ps6-19) and engineered C. propionicum propionyl-CoA transferase (PctCp), respectively, were expressed in E. coli W to construct key metabolic pathway to produce poly(3-hydroxybutyrate-co-lactate) [P(3HB-co-LA)]. The recombinant E. coli W expressing the phaC1437 gene and the pct540 gene could synthesize P(3HB-co-13mol%LA) up to the polymer content of 31.3 wt% when it was cultured in chemically defined MR medium containing 20 g/L of sucrose and 2 g/L of sodium 3-hydroxybutyrate. When Ralstonia eutropha phaAB genes were additionally expressed to provide 3-hydroxybutyrate-CoA (3HB-CoA) from sucrose, P(3HB-co-16mol%LA) could be synthesized from sucrose as a sole carbon source without supplement of sodium 3-hydroxybutyrate in culture medium, but the PHA content was decreased to 12.2 wt%. The molecular weight of P(3HB-co-16mol%LA) synthesized in E. coli W using sucrose as carbon source were 1.53×104 (Mn) and 2.78×104 (Mw), respectively, which are not different from those that have previously been reported by other recombinant E. coli strains. Engineered E. coli strains developed in this study should be useful for the production of lactate-containing PHAs from sucrose, one of the most abundant and least expensive carbon sources.
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KEYWORD
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Escherichia coli, Sucrose, Polyhydroxyalkanoate (PHA), Lactate-containing PHA
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