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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >A Microbial Factory For Lactate-based Polyesters Using A Lactate-polymerizing Enzyme
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A Microbial Factory For Lactate-based Polyesters Using A Lactate-polymerizing Enzyme

机译：使用乳酸聚合酶的乳酸基聚酯微生物工厂

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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 poly-hydroxyalkanoate (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 electrophore-sis/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 copoly-ester, P(6 mol% LA-co-94 mol% 3HB), with a number-average molecular weight of 1.9 × 10~5, as revealed by gel permeation chromatography, gas chromatography/MS, and NMR.
机译：聚乳酸（PLA）是通过化学生物过程，在微生物发酵提供的乳酸（LA）的金属催化剂介导的化学聚合的基础上，通过化学生物过程合成的，作为具有代表性的生物基聚酯。为了建立用于合成LA基聚酯的一步法微生物工艺，我们基于PHA单体成分（尤其是PHA单体成分）这一事实，探讨了聚羟基链烷酸酯（PHA）合酶是否对LA辅酶A（CoA）表现出聚合活性。 3-羟基丁酸酯（3HB）在结构上类似于LA。通过先前开发的两阶段体外聚合系统，发现一种工程化的PHA合酶可作为候选物。构建了具有CoA转移酶基因的产LA-CoA的大肠杆菌菌株，并通过毛细管电泳-MS / MS分析证明了LA-CoA的产生。接下来，当将工程改造的PHA合酶基因引入所得重组菌株中时，我们证实了掺入LA的共聚酯P（6 mol％LA-co-94 mol％3HB）的一步合成。通过凝胶渗透色谱法，气相色谱/ MS和NMR显示的平均分子量为1.9×10-5。

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