Optogenetic Fine-Tuning of Sus scrofa Basic Fibroblast Growth Factor Expression in Escherichia coli
文献类型： 外文期刊
作者： Meng, Fanqiang 1 ; Xu, Zhimin 2 ; Fan, Xia 1 ; Wang, Zhisheng 3 ; Zhou, Libang 1 ;
作者机构： 1.Nanjing Agr Univ, Coll Food Sci & Technol, Nanjing 210095, Peoples R China
2.Shandong Vocat Anim Sci & Vet Coll, Dept Food & Drug Engn, Weifang 261061, Peoples R China
3.Jiangsu Acad Agr Sci, Natl Res Ctr Engn & Technol Vet Biol, Nanjing 210014, Peoples R China
关键词：
optogenetic; EL222; bFGF; blue light;
期刊名称：FERMENTATION-BASEL （ 影响因子：3.3； 五年影响因子：3.5 ）
ISSN：
年卷期： 2024 年 10 卷 12 期
页码：
收录情况： SCI
摘要： Basic fibroblast growth factor (bFGF) is a crucial protein with diverse applications in biotechnology and medicine. This study aims to investigate the use of EL222-based optogenetic control systems to fine-tune the expression of porcine (Sus scrofa) bFGF in Escherichia coli. The bioactivity and the productivity of blue light-induced bFGF were demonstrated to be comparable to those achieved using a conventional T7-expression system. Secondly, through systematic optimization of regulatory elements, optimal expression of bFGF was achieved using a medium-strength promoter for EL222 expression, a strong RBS upstream of the bFGF gene, and an optimized C120 configuration within the blue light-inducible promoter. Moreover, various parameters of blue light illumination during fermentation were investigated, including initial cell density, light intensity, illumination duration, and pulsed illumination patterns. The results identified optimal conditions for maximizing bFGF yield in E. coli, specifically an initial OD600 of 0.6, 800 lux blue light intensity, and 8 h total illumination in a 2 h on/off pattern. Overall, this successful implementation of optogenetically controlled bFGF expression in E. coli serves as a proof-of-concept for light-responsive systems in industrial biotechnology, highlighting the potential of optogenetic control for biologically active protein production.
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