Microbial Secretion Platform for 3-Hydroxybutyrate Oligomer and Its End-Capped Forms Using Chain Transfer Reaction-Mediated Polyhydroxyalkanoate Synthases.,Biotechnology Journal

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Microbial Secretion Platform for 3-Hydroxybutyrate Oligomer and Its End-Capped Forms Using Chain Transfer Reaction-Mediated Polyhydroxyalkanoate Synthases.
Biotechnology Journal ( IF 3.2 ) Pub Date : 2019-11-21 , DOI: 10.1002/biot.201900201
Yuki Miyahara _{1,

2} , Ayaka Hiroe _{1,

2} , Takeharu Tsuge _{2,

3} , Seiichi Taguchi _{1,

4}

Affiliation

The biodegradable polyester 3-hydroxybutyrate (3HB) polymer [P(3HB)] is intracellularly synthesized and accumulated in recombinant Escherichia coli. In this study, native polyhydroxyalkanoate (PHA) synthases are used to attempt to microbially secrete 3HB homo-oligomers (3HBOs), which are widely distributed in nature as physiologically active substances. High secretory production is observed, especially for the two PHA synthases from Aeromonas caviae and Bacillus cereus YB4. Surprisingly, an ethyl ester at the carboxy terminus (ethyl ester form) of 3HBOs is identified for most of the PHA synthases tested. Next, 3HBOs with a functional carboxyl group (carboxyl form of 3HBO) are obtained by using the alcohol dehydrogenase gene (adhE)-deficient mutant strain, suggesting that the endogenous ethanol produced in E. coli acts as a chain transfer (CT) agent in the generation of 3HBOs. Furthermore, an in vitro polymerization assay reveals that CT agents such as ethanol and free 3HB are involved in the generation of ethyl ester and carboxyl form of 3HBO, respectively. The microbial platform established herein allows the secretion of 3HBOs with desirable end structures by supplementation with various CT agents. The obtained 3HBOs and their end-capped forms may be used as physiologically active substances and building blocks for polymeric materials.

中文翻译：

使用链转移反应介导的聚羟基链烷酸酯合成酶为3-羟基丁酸低聚物及其封端形式的微生物分泌平台。

可生物降解的聚酯3-羟基丁酸酯（3HB）聚合物[P（3HB）]在细胞内合成并积累在重组大肠杆菌中。在这项研究中，天然的聚羟基链烷酸酯（PHA）合成酶用于尝试分泌3HB均聚物（3HBOs），它们以生理活性物质的形式广泛分布于自然界。观察到高分泌产生，特别是对于来自鼠气单胞菌和蜡状芽孢杆菌YB4的两种PHA合酶。出乎意料的是，对于大多数测试的PHA合成酶，都鉴定出3HBOs羧基末端的乙酯（乙酯形式）。接下来，通过使用醇脱氢酶基因（adhE）缺陷型突变株获得具有功能性羧基的3HBO（羧基形式的3HBO），这表明E中产生了内源性乙醇。大肠杆菌在3HBO的产生中充当链转移（CT）剂。此外，体外聚合试验表明，CT试剂（例如乙醇和游离的3HB）分别参与了3HBO的乙酯和羧基形式的生成。本文建立的微生物平台可通过补充各种CT试剂来分泌具有所需末端结构的3HBO。所获得的3HBO及其封端形式可以用作生理活性物质和高分子材料的结构单元。本文建立的微生物平台可通过补充各种CT试剂来分泌具有所需末端结构的3HBO。所获得的3HBO及其封端形式可以用作生理活性物质和高分子材料的结构单元。本文建立的微生物平台可通过补充各种CT试剂来分泌具有所需末端结构的3HBO。所获得的3HBO及其封端形式可以用作生理活性物质和高分子材料的结构单元。

更新日期：2019-11-22

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