Applied Microbiology and Biotechnology

, Volume 85, Issue 4, pp 949–954

Biosynthesis of novel terpolymers poly(lactate-co-3-hydroxybutyrate-co-3-hydroxyvalerate)s in lactate-overproducing mutant Escherichia coli JW0885 by feeding propionate as a precursor of 3-hydroxyvalerate

  • Fumi Shozui
  • Ken’ichiro Matsumoto
  • Takanori Nakai
  • Miwa Yamada
  • Seiichi Taguchi
Biotechnological Products and Process Engineering

Abstract

Novel lactate (LA)-based terpolymers, P[LA-co-3-hydroxybutyrate(3HB)-co-3-hydroxyvalerate(3HV)]s (PLBVs), were produced in LA-overproducing mutant, Escherichia coli JW0885, which was found to be a superior host for the efficient production of LA-based polyesters. Recombinant E. coli JW0885 harboring the genes encoding LA-polymerizing enzyme (Ser325Thr/Gln481Lys mutant of polyhydroxyalkanoate synthase from Pseudomonas sp. 61-3) and three monomer supplying enzymes [propionyl-CoA transferase, β-ketothiolase, and nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH)-dependent acetoacetyl-CoA reductase] was aerobically grown on glucose with feeding of propionate as a precursor of 3-hydroxyvaleryl-CoA (3HV-CoA). Gas chromatography and nuclear magnetic resonance (NMR) analyses revealed that polymers accumulated in the cells were composed of LA, 3HB, and 3HV units, thus being identified as terpolymers, PLBVs. In addition, 1H-NMR analysis suggested the existence of LA-3HV sequence in the terpolymer. When 100 mg/l of sodium propionate was added into the medium, 3HV fraction in the terpolymer linearly reached up to 7.2 mol%, while LA fraction was inversely decreased. This phenomenon could be due to the change in metabolic fluxes of lactyl-CoA (LA-CoA) and 3HV-CoA depending on the concentration of propionate fed into the medium.

Keywords

Microbial polymer production Copolymer Poly(LA-co-3HB-co-3HV) Substrate specificity Metabolic engineering PhaC 

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Fumi Shozui
    • 1
  • Ken’ichiro Matsumoto
    • 1
  • Takanori Nakai
    • 1
  • Miwa Yamada
    • 1
  • Seiichi Taguchi
    • 1
  1. 1.Division of Biotechnology and Macromolecular Chemistry, Graduate School of EngineeringHokkaido UniversitySapporoJapan

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