Flux optimization using multiple promoters in Halomonas bluephagenesis as a model chassis of the next generation industrial biotechnology.

Yueyuan Ma, Jian-Wen Ye, Yina Lin, Xueqing Yi, Xuan Wang, Huan Wang, Ruiyan Huang, Fuqing Wu, Qiong Wu, Xu Liu, Guo-Qiang Chen

Metabolic engineering 2024 Jan

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Predictability and robustness are challenges for bioproduction because of the unstable intracellular synthetic activities. With the deeper understanding of the gene expression process, fine-tuning has become a meaningful tool for biosynthesis optimization. This study characterized several gene expression elements and constructed a multiple inducible system that responds to ten different small chemical inducers in halophile bacterium Halomonas bluephagenesis. Genome insertion of regulators was conducted for the purpose of gene cluster stabilization and regulatory plasmid simplification. Additionally, dynamic ranges of the multiple inducible systems were tuned by promoter sequence mutations to achieve diverse scopes for high-resolution gene expression control. The multiple inducible system was successfully employed to precisely control chromoprotein expression, lycopene and poly-3-hydroxybutyrate (PHB) biosynthesis, resulting in colorful bacterial pictures, optimized cell growth, lycopene and PHB accumulation. This study demonstrates a desirable approach for fine-tuning of rational and efficient gene expressions, displaying the significance for metabolic pathway optimization. Copyright © 2023. Published by Elsevier Inc.

Citation

Yueyuan Ma, Jian-Wen Ye, Yina Lin, Xueqing Yi, Xuan Wang, Huan Wang, Ruiyan Huang, Fuqing Wu, Qiong Wu, Xu Liu, Guo-Qiang Chen. Flux optimization using multiple promoters in Halomonas bluephagenesis as a model chassis of the next generation industrial biotechnology. Metabolic engineering. 2024 Jan;81:249-261