A Microbial Factory for Lactate-based Polyesters Using a Lactate-polymerizing Enzyme

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2008 Nov 4

PMID 18978031

Citations 56

Authors

Seiichi Taguchi

Miwa Yamada

Kenichiro Matsumoto

Kenji Tajima

Yasuharu Satoh

Masanobu Munekata

Katsuhiro Ohno

Katsunori Kohda

Takashi Shimamura

Hiromi Kambe

Shusei Obata

Affiliations

Soon will be listed here.

Abstract

Polylactate (PLA) is synthesized as a representative bio-based polyester by the chemo-bio process on the basis of metal catalyst-mediated chemical polymerization of lactate (LA) supplied by microbial fermentation. To establish the one-step microbial process for synthesis of LA-based polyesters, we explored whether polyhydroxyalkanoate (PHA) synthase would exhibit polymerizing activity toward a LA-coenzyme A (CoA), based on the fact that PHA monomeric constituents, especially 3-hydroxybutyrate (3HB), are structurally analogous to LA. An engineered PHA synthase was discovered as a candidate by a two-phase in vitro polymerization system previously developed. An LA-CoA producing Escherichia coli strain with a CoA transferase gene was constructed, and the generation of LA-CoA was demonstrated by capillary electrophoresis/MS analysis. Next, when the engineered PHA synthase gene was introduced into the resultant recombinant strain, we confirmed the one-step biosynthesis of the LA-incorporated copolyester, P(6 mol% LA-co-94 mol% 3HB), with a number-average molecular weight of 1.9 x 10(5), as revealed by gel permeation chromatography, gas chromatography/MS, and NMR.

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References

Matsusaki H, Manji S, Taguchi K, Kato M, Fukui T, Doi Y . 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. 1998; 180(24):6459-67. PMC: 107745. DOI: 10.1128/JB.180.24.6459-6467.1998. View

Taguchi S, Doi Y . Evolution of polyhydroxyalkanoate (PHA) production system by "enzyme evolution": successful case studies of directed evolution. Macromol Biosci. 2004; 4(3):146-56. DOI: 10.1002/mabi.200300111. View

Yuan W, Jia Y, Tian J, Snell K, Muh U, Sinskey A . Class I and III polyhydroxyalkanoate synthases from Ralstonia eutropha and Allochromatium vinosum: characterization and substrate specificity studies. Arch Biochem Biophys. 2001; 394(1):87-98. DOI: 10.1006/abbi.2001.2522. View

Selmer T, Willanzheimer A, Hetzel M . Propionate CoA-transferase from Clostridium propionicum. Cloning of gene and identification of glutamate 324 at the active site. Eur J Biochem. 2002; 269(1):372-80. DOI: 10.1046/j.0014-2956.2001.02659.x. View

Steinbuchel A, Hein S . Biochemical and molecular basis of microbial synthesis of polyhydroxyalkanoates in microorganisms. Adv Biochem Eng Biotechnol. 2001; 71:81-123. DOI: 10.1007/3-540-40021-4_3. View

Matsumoto K, Nakae S, Taguchi K, Matsusaki H, Seki M, Doi Y . Biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyalkanoates) copolymer from sugars by recombinant Ralstonia eutropha harboring the phaC1Ps and the phaGPs genes of Pseudomonas sp. 61-3. Biomacromolecules. 2001; 2(3):934-9. DOI: 10.1021/bm0155367. View

Takase K, Matsumoto K, Taguchi S, Doi Y . 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. 2004; 5(2):480-5. DOI: 10.1021/bm034323+. View

Takase K, Taguchi S, Doi Y . Enhanced synthesis of poly(3-hydroxybutyrate) in recombinant Escherichia coli by means of error-prone PCR mutagenesis, saturation mutagenesis, and in vitro recombination of the type II polyhydroxyalkanoate synthase gene. J Biochem. 2003; 133(1):139-45. DOI: 10.1093/jb/mvg015. View

Peoples O, Sinskey A . Poly-beta-hydroxybutyrate biosynthesis in Alcaligenes eutrophus H16. Characterization of the genes encoding beta-ketothiolase and acetoacetyl-CoA reductase. J Biol Chem. 1989; 264(26):15293-7. View

10.

Soga T, Ueno Y, Naraoka H, Matsuda K, Tomita M, Nishioka T . Pressure-assisted capillary electrophoresis electrospray ionization mass spectrometry for analysis of multivalent anions. Anal Chem. 2003; 74(24):6224-9. DOI: 10.1021/ac0202684. View

11.

Rehm B . Polyester synthases: natural catalysts for plastics. Biochem J. 2003; 376(Pt 1):15-33. PMC: 1223765. DOI: 10.1042/BJ20031254. View

12.

Auras R, Harte B, Selke S . An overview of polylactides as packaging materials. Macromol Biosci. 2004; 4(9):835-64. DOI: 10.1002/mabi.200400043. View

13.

ELSDEN S, Gilchrist F, Lewis D, Volcani B . Properties of a fatty acid forming organism isolated from the rumen of sheep. J Bacteriol. 1956; 72(5):681-9. PMC: 357977. DOI: 10.1128/jb.72.5.681-689.1956. View

14.

Stubbe J, Tian J . Polyhydroxyalkanoate (PHA) hemeostasis: the role of PHA synthase. Nat Prod Rep. 2003; 20(5):445-57. DOI: 10.1039/b209687k. View

15.

Vieira J, Messing J . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985; 33(1):103-19. DOI: 10.1016/0378-1119(85)90120-9. View

16.

Nomura C, Taguchi S . PHA synthase engineering toward superbiocatalysts for custom-made biopolymers. Appl Microbiol Biotechnol. 2006; 73(5):969-79. DOI: 10.1007/s00253-006-0566-4. View

17.

Satoh Y, Tajima K, Tannai H, Munekata M . Enzyme-catalyzed poly(3-hydroxybutyrate) synthesis from acetate with CoA recycling and NADPH regeneration in Vitro. J Biosci Bioeng. 2005; 95(4):335-41. DOI: 10.1016/s1389-1723(03)80064-6. View

18.

Arai Y, Nakashita H, Suzuki Y, Kobayashi Y, Shimizu T, Yasuda M . Synthesis of a novel class of polyhydroxyalkanoates in Arabidopsis peroxisomes, and their use in monitoring short-chain-length intermediates of beta-oxidation. Plant Cell Physiol. 2002; 43(5):555-62. DOI: 10.1093/pcp/pcf068. View