Zong Hong, Zhang Liya, Cheng Yiwen, Sheng Zhiying, Zhuge Bin, Lu Xinyao
School of Biotechnology and Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, China.
Lab of Industrial Microorganism & Research and Design Center for Polyols, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
ACS Synth Biol. 2025 Jan 17;14(1):273-284. doi: 10.1021/acssynbio.4c00798. Epub 2025 Jan 5.
DegSU quorum sensing (QS) system enables autoinducible expression of recombinant proteins in . However, insufficient promoter strength and a complex regulatory circuit limit its practical application. Here, the QS-responsive promoter P was modified by core region mutation, upstream truncation, and addition of activating binding sites, yielding P 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 by knocking out competitive target genes and , operons and , introducing variants and 36, 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.
DegSU群体感应(QS)系统能够在……中实现重组蛋白的自诱导表达。然而,启动子强度不足和复杂的调控回路限制了其实际应用。在此,通过核心区域突变、上游截短和添加激活结合位点对QS响应启动子P进行了修饰,得到强度提高了118.3%的P。开发了一个数学模型,从全面的角度准确量化调控过程。在该模型的指导下,通过敲除竞争性靶基因……和……、操纵子……和……、引入变体……和36,并将调控强度提高84.0%,在一个稳健的……中对DegSU QS系统进行了进一步优化。乙偶姻滴度提高了52.5%,细胞外羧肽酶活性提高了65.9%,验证了本研究的工业价值。总体而言,本研究解决了DegSU QS系统在实际应用中的局限性,并证明了其在高水平重组蛋白生产方面的潜力。