Production and molecular weight variation of poly-γ-glutamic acid using a recombinant Bacillus subtilis with various Pgs-component ratios.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY Accounts of Chemical Research Pub Date : 2024-09-20 DOI:10.1093/bbb/zbae093

Kazuhisa Sawada, Hiroshi Hagihara, Yasushi Takimura, Masakazu Kataoka

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Abstract

Poly-γ-glutamic acid (PGA) has been of interest as a sustainable biopolymer in industrial applications. PGA biosynthesis in Bacillus subtilis is catalyzed by a transmembrane protein complex comprising PgsB, PgsC, and PgsA. To determine the Pgs component responsible for PGA overproduction, we constructed recombinants in which the promoter of the host-derived pgs gene was replaced with another host-derived gene promoter. These recombinants were then transformed using high-copy-number plasmids with various pgs-gene combinations to enhance Pgs component in different ratios. Subsequently, PGA production was investigated in batch cultures with l-glutamate supplemented medium. The recombinant strain enhanced with pgsB alone significantly overproduced PGA (maximum production 35.8 g/L) than either the pgsC- or pgsA-enhanced strain. The molecular weight of the PGA produced with the pgsB-enhanced strain was also greater than that for the pgsC- or pgsA-enhanced strain (approximately 10-fold). Hence, PgsB enhancement alone contributes to PGA overproduction with increased molecular weight.

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利用重组枯草芽孢杆菌以不同的 pgs-组分比例生产聚-γ-谷氨酸并改变其分子量。

聚γ-谷氨酸（PGA）作为一种可持续生物聚合物，在工业应用中一直备受关注。枯草芽孢杆菌中的 PGA 生物合成是由 PgsB、PgsC 和 PgsA 组成的跨膜蛋白复合物催化的。为了确定导致 PGA 过量产生的 Pgs 成分，我们构建了重组子，其中宿主衍生 pgs 基因的启动子被另一个宿主衍生基因的启动子取代。然后，使用带有不同 pgs 基因组合的高拷贝数质粒转化这些重组子，以不同比例增强 Pgs 成分。随后，在补充了 l-谷氨酸的培养基中进行批量培养，研究 PGA 的生产情况。与 pgsC 或 pgsA 增强菌株相比，仅用 pgsB 增强的重组菌株明显多产 PGA（最大产量为 35.8 gL-1）。pgsB 增强菌株产生的 PGA 的分子量也比 pgsC 或 pgsA 增强菌株大（约 10 倍）。因此，仅 PgsB 增强就会导致 PGA 生产过剩并增加分子量。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.