Zhang Jinfang, Zhu Baoyue, Li Xinyue, Xu Xiaojian, Li Dengke, Zeng Fang, Zhou Cuixia, Liu Yihan, Li Yu, Lu Fuping
Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, the College of Biotechnology, Tianjin University of Science and Technology, Tianjin, China.
School of Biology and Brewing Engineering, Taishan University, Taian, China.
Front Bioeng Biotechnol. 2022 Apr 14;10:866066. doi: 10.3389/fbioe.2022.866066. eCollection 2022.
is a generally recognized as safe (GRAS) microorganism that presents great potential for the production of heterologous proteins. In this study, we performed genomic and comparative transcriptome to investigate the critical modular in on the production of heterologous alkaline proteases (AprE). After investigation, it was concluded that the key modules affecting the production of alkaline protease were the sporulation germination module (Module I), extracellular protease synthesis module (Module II), and extracellular polysaccharide synthesis module (Module III) in . In Module I, AprE yield for mutant BA Δ was 25.3% greater than that of BA Δ. Combining Module I synergistically with mutation of extracellular proteases in Module II significantly increased AprE production by 36.1% compared with production by BA Δ. In Module III, the mutation of genes controlling extracellular polysaccharides reduced the viscosity and the accumulation of sediment, and increased the rate of dissolved oxygen in fermentation. Moreover, AprE production was 39.6% higher than in BA Δ when Modules I, II and III were engineered in combination. This study provides modular engineering strategies for the modification of for the production of alkaline proteases.
是一种普遍认为安全（GRAS）的微生物，在生产异源蛋白方面具有巨大潜力。在本研究中，我们进行了基因组和比较转录组分析，以研究在生产异源碱性蛋白酶（AprE）方面的关键模块。经过研究得出结论，影响碱性蛋白酶生产的关键模块是中的孢子形成萌发模块（模块I）、细胞外蛋白酶合成模块（模块II）和细胞外多糖合成模块（模块III）。在模块I中，突变体BA Δ的AprE产量比BA Δ高25.3%。将模块I与模块II中细胞外蛋白酶的突变协同结合，与BA Δ的产量相比，AprE产量显著提高了36.1%。在模块III中，控制细胞外多糖的基因突变降低了粘度和沉淀物的积累，并提高了发酵中的溶解氧速率。此外，当模块I、II和III组合改造时，AprE产量比BA Δ高39.6%。本研究为改造用于生产碱性蛋白酶提供了模块工程策略。
