A microbial factory for lactate-based polyesters using a lactate-polymerizing enzyme

被引：237

作者：

Taguchi, Seiichi ^{[1
]}

Yamadaa, Miwa ^{[1
]}

Matsumoto, Ken'ichiro ^{[1
]}

Tajima, Kenji ^{[1
]}

Satoh, Yasuharu ^{[1
]}

Munekata, Masanobu ^{[1
]}

Ohno, Katsuhiro ^{[2
]}

Kohda, Katsunori ^{[2
]}

Shimamura, Takashi ^{[2
]}

Kambe, Hiromi

Obata, Shusei ^{[3
]}

机构：

[1] Hokkaido Univ, Grad Sch Engn, Div Biotechnol & Mol Chem, Kita Ku, Sapporo, Hokkaido 0608628, Japan

[2] Toyota Cent Res & Dev Labs Inc, Dept Mat, Sustainable Mat Div, Biotechnol Lab, Aichi 4801192, Japan

[3] Toyota Motor Co Ltd, Future Project Div, Bio Res Lab, Aichi 4718572, Japan

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2008年 / 105卷 / 45期

关键词：

lactate coenzyme A; polyhydroxyalkanoate synthase; substrate specificity; CoA transferase; enzyme engineering;

D O I：

10.1073/pnas.0805653105

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：17323 / 17327

页数：5

共 22 条

[1] Physical properties and enzymatic degradability of copolymers of (R)-3-hydroxybutyric acid and (S,S)-lactide [J].