Design-build-test of recombinant Bacillus subtilis chassis cell by lifespan engineering for robust bioprocesses

IF 4.4 2区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Synthetic and Systems Biotechnology Pub Date : 2024-04-11 DOI:10.1016/j.synbio.2024.04.004
Kexin Ren , Qiang Wang , Jianghua Chen , Hengwei Zhang , Zhoule Guo , Meijuan Xu , Zhiming Rao , Xian Zhang
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Abstract

Microbial cell factories utilize renewable raw materials for industrial chemical production, providing a promising path for sustainable development. Bacillus subtilis is widely used in industry for its food safety properties, but challenges remain in the limitations of microbial fermentation. This study proposes a novel strategy based on lifespan engineering to design robust B. subtilis chassis cells to supplement traditional metabolic modification strategies that can alleviate cell autolysis, tolerate toxic substrates, and get a higher mass transfer efficiency. The modified chassis cells could produce high levels of l-glutaminase, and tolerate hydroquinone to produce α-arbutin efficiently. In a 5 L bioreactor, the l-glutaminase enzyme activity of the final strain CRE15TG was increased to 2817.4 ± 21.7 U mL−1, about 1.98-fold compared with that of the wild type. The α-arbutin yield of strain CRE15A was increased to 134.7 g L−1, about 1.34-fold compared with that of the WT. To our knowledge, both of the products in this study performed the highest yields reported so far. The chassis modification strategy described in this study can Improve the utilization efficiency of chassis cells, mitigate the possible adverse effects caused by excessive metabolic modification of engineered strains, and provide a new idea for the future design of microbial cell factories.

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通过寿命工程设计-构建-测试重组枯草芽孢杆菌底盘细胞,实现稳健的生物过程
微生物细胞工厂利用可再生原料生产工业化学品,为可持续发展提供了一条前景广阔的道路。枯草芽孢杆菌因其食品安全特性被广泛应用于工业领域,但微生物发酵的局限性仍是挑战。本研究提出了一种基于生命周期工程学的新策略,以设计稳健的枯草芽孢杆菌底盘细胞来补充传统的代谢改造策略,从而减轻细胞自溶、耐受有毒底物并获得更高的传质效率。改造后的底盘细胞能产生高水平的 l-谷氨酰胺酶,并能耐受对苯二酚,从而高效生产 α-熊果苷。在 5 L 生物反应器中,最终菌株 CRE15TG 的 l-谷氨酰胺酶活性增至 2817.4 ± 21.7 U mL-1,是野生型的 1.98 倍。菌株 CRE15A 的 α-arbutin 产量增至 134.7 g L-1,约为 WT 的 1.34 倍。据我们所知,本研究中的两种产品都是迄今为止所报道的产量最高的。本研究中描述的底盘改造策略可以提高底盘细胞的利用效率,减轻工程菌株过度代谢改造可能带来的不良影响,并为未来微生物细胞工厂的设计提供了新思路。
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来源期刊
Synthetic and Systems Biotechnology
Synthetic and Systems Biotechnology BIOTECHNOLOGY & APPLIED MICROBIOLOGY-
CiteScore
6.90
自引率
12.50%
发文量
90
审稿时长
67 days
期刊介绍: Synthetic and Systems Biotechnology aims to promote the communication of original research in synthetic and systems biology, with strong emphasis on applications towards biotechnology. This journal is a quarterly peer-reviewed journal led by Editor-in-Chief Lixin Zhang. The journal publishes high-quality research; focusing on integrative approaches to enable the understanding and design of biological systems, and research to develop the application of systems and synthetic biology to natural systems. This journal will publish Articles, Short notes, Methods, Mini Reviews, Commentary and Conference reviews.
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