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

IF 1.4 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Bioscience, Biotechnology, and Biochemistry Pub Date : 2024-09-20 DOI:10.1093/bbb/zbae093

Kazuhisa Sawada, Hiroshi Hagihara, Yasushi Takimura, Masakazu Kataoka

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引用次数: 0

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。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Bioscience, Biotechnology, and Biochemistry 生物-生化与分子生物学

CiteScore

3.50

自引率

0.00%

发文量

183

审稿时长

1 months

期刊介绍： Bioscience, Biotechnology, and Biochemistry publishes high-quality papers providing chemical and biological analyses of vital phenomena exhibited by animals, plants, and microorganisms, the chemical structures and functions of their products, and related matters. The Journal plays a major role in communicating to a global audience outstanding basic and applied research in all fields subsumed by the Japan Society for Bioscience, Biotechnology, and Agrochemistry (JSBBA).