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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

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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.

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References

  • Aldor IS, Keasling JD (2003) Process design for microbial plastic factories: metabolic engineering of polyhydroxyalkanoates. Curr Opin Biotechnol 14:475–483

    Article  CAS  Google Scholar 

  • Anderson AJ, Dawes EA (1990) Occurrence, metabolism, metabolic role, and industrial uses of bacterial polyhydroxyalkanoates. Microbiol Rev 54:450–472

    CAS  Google Scholar 

  • Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006) Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection. Mol Syst Biol 2:1–11

    Article  Google Scholar 

  • Blank L, Green J, Guest JR (2002) AcnC of Escherichia coli is a 2-methylcitrate dehydratase (PrpD) that can use citrate and isocitrate as substrates. Microbiology 148:133–146

    CAS  Google Scholar 

  • Doi Y, Kunioka M, Nakamura Y, Soga K (1986) Nuclear magnetic resonance studies on poly(beta-hydroxybutyrate) and a copolyester of beta-hydroxybutyrate and beta-hydroxyvalerate isolated from Alcaligenes eutrophus H16. Macromolecules 19:2860–2864

    Article  CAS  Google Scholar 

  • Elsden SR, Gilchrist FM, Lewis D, Volcani BE (1956) Properties of a fatty acid forming organism isolated from the rumen of sheep. J Bacteriol 72:681–689

    Article  CAS  Google Scholar 

  • Holmes PA (1985) Applications of PHB—a microbially produced biodegradable thermoplastic. Phys Technol 16:32–36

    Article  CAS  Google Scholar 

  • Jia Z, Zhang K, Tan J, Han C, Dong L, Yang Y (2009) Crystallization behavior and mechanical properties of crosslinked plasticized poly(L-lactic acid). J Appl Polym Sci 111:1530–1539

    Article  CAS  Google Scholar 

  • Kato M, Bao HJ, Kang CK, Fukui T, Doi Y (1996) Production of a novel copolyester of 3-hydroxybutyric acid and medium-chain-length 3-hydroxyalkanoic acids by Pseudomonas sp. 61–3 from sugars. Appl Microbiol Biotechnol 45:363–370

    Article  CAS  Google Scholar 

  • Kulinski Z, Piorkowska E, Gadzinowska K, Stasiak M (2006) Plasticization of poly(L-lactide) with poly(propylene glycol). Biomacromolecules 7:2128–2135

    Article  CAS  Google Scholar 

  • Matsumoto K, Murata T, Nagao R, Nomura CT, Arai S, Arai Y, Takase K, Nakashita H, Taguchi S, Shimada H (2009) Production of short-chain-length/medium-chain-length polyhydroxyalkanoate (PHA) copolymer in the plastid of Arabidopsis thaliana using an engineered 3-ketoacyl-acyl carrier protein synthase III. Biomacromolecules 10:686–690

    Article  CAS  Google Scholar 

  • Matsusaki H, Abe H, Doi Y (2000) Biosynthesis and properties of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) by recombinant strains of Pseudomonas sp. 61–3. Biomacromolecules 1:17–22

    Article  CAS  Google Scholar 

  • Matsusaki H, Manji S, Taguchi K, Kato M, Fukui T, Doi Y (1998) Cloning and molecular analysis of the poly(3-hydroxybutyrate) and poly(3-hydroxybutyrate-co-3-hydroxyalkanoate) biosynthesis genes in Pseudomonas sp. strain 61–3. J Bacteriol 180:6459–6467

    CAS  Google Scholar 

  • Schubert P, Steinbüchel A, Schlegel HG (1988) Cloning of the Alcaligenes eutrophus genes for synthesis of poly-beta-hydroxybutyric acid (PHB) and synthesis of PHB in Escherichia coli. J Bacteriol 170:5837–5847

    CAS  Google Scholar 

  • Schweiger G, Buckel W (1984) On the dehydration of (R)-lactate in the fermentation of alanine to propionate by Clostridium propionicum. FEBS letters 171:79–84

    Article  CAS  Google Scholar 

  • Shirahama H, Tanaka A, Yasuda H (2002) Highly biodegradable copolymers composed of chiral depsipeptide and L-lactide units with favorable physical properties. J Polym Sci A Polym Chem 40:302–316

    Article  CAS  Google Scholar 

  • Slater S, Gallaher T, Dennis D (1992) Production of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) in a recombinant Escherichia coli strain. Appl Environ Microbiol 58:1089–1094

    CAS  Google Scholar 

  • Sudesh K, Abe H, Doi Y (2000) Synthesis, structure and properties of polyhydroxyalkanoates: biological polyester. Prog Polym Sci 25:1503–1555

    Article  CAS  Google Scholar 

  • Taguchi S, Yamada M, Matsumoto K, Tajima K, Satoh Y, Munekata M, Ohno K, Kohda K, Shimamura T, Kambe H, Obata S (2008) A microbial factory for lactate-based polyesters using a lactate-polymerizing enzyme. Proc Natl Acad Sci U S A 105:17323–17327

    Article  CAS  Google Scholar 

  • Takase K, Matsumoto K, Taguchi S, Doi Y (2004) Alteration of substrate chain-length specificity of type II synthase for polyhydroxyalkanoate biosynthesis by in vitro evolution: in vivo and in vitro enzyme assays. Biomacromolecules 5:480–485

    Article  CAS  Google Scholar 

  • Tsuji H (2002) Polylactide. In: Doi Y, Steinbüchel A (eds) Biopolymers. Wiley-VCH, Weinheim, pp 129–177

    Google Scholar 

  • Yamada M, Matsumoto K, Nakai T, Taguchi S (2009) Microbial production of lactate-enriched poly[(R)-lactate-co-(R)-3-hydroxybutyrate] with novel thermal properties. Biomacromolecules 10:677–681

    Article  CAS  Google Scholar 

  • Yim KS, Lee SY, Chang HN (1996) Synthesis of poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) by recombinant Escherichia coli. Biotechnol Bioeng 49:495–503

    Article  CAS  Google Scholar 

  • Zhu J, Shimizu K (2005) Effect of a single-gene knockout on the metabolic regulation in Escherichia coli for D-lactate production under microaerobic condition. Metab Eng 7:104–115

    Article  CAS  Google Scholar 

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Acknowledgements

We thank NBRP (NIG, Japan): E. coli for providing E. coli JW0885. This work was supported by funding from the following sources: Grant-in-Aid for Scientific Research of Japan (no. 70216828) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (to S. Taguchi) and the Global Center of Excellence Program (project no. B01: Catalysis as the Basis for Innovation in Materials Science) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan.

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  1. Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, N13W8, Kita-ku, Sapporo, 060-8628, Japan

    Fumi Shozui, Ken’ichiro Matsumoto, Takanori Nakai, Miwa Yamada & Seiichi Taguchi

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  1. Fumi Shozui
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  2. Ken’ichiro Matsumoto
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  3. Takanori Nakai
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  4. Miwa Yamada
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  5. Seiichi Taguchi
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Correspondence to Seiichi Taguchi.

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Shozui, F., Matsumoto, K., Nakai, T. et al. 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. Appl Microbiol Biotechnol 85, 949–954 (2010). https://doi.org/10.1007/s00253-009-2100-y

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