Efficient Autoinducible Expression of Recombinant Proteins via the DegSU Quorum Sensing System in a Robust Bacillus subtilis.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS ACS Synthetic Biology Pub Date : 2025-01-05 DOI:10.1021/acssynbio.4c00798

Hong Zong, Liya Zhang, Yiwen Cheng, Zhiying Sheng, Bin Zhuge, Xinyao Lu

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Abstract

DegSU quorum sensing (QS) system enables autoinducible expression of recombinant proteins in Bacillus subtilis . However, insufficient promoter strength and a complex regulatory circuit limit its practical application. Here, the QS-responsive promoter P_aprE was modified by core region mutation, upstream truncation, and addition of activating binding sites, yielding P_E742 with a 118.3% increase in strength. A mathematical model was developed to accurately quantify the regulatory process from a comprehensive perspective. Guided by this model, the DegSU QS system was further optimized in a robust B. subtilis by knocking out competitive target genes sacB and amyE, operons pgs and srfA, introducing variants degU^L113F and degQ36^Hy, and increasing regulatory strength by 84.0%. A 52.5% increase in acetoin titer and a 65.9% increase in extracellular carboxypeptidase activity validated the industrial value of this study. Overall, this study addresses the limitations of the DegSU QS system in practical application and demonstrates its potential for high-level recombinant protein production.

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.