Improvement surfactin production by substitution of promoters in Bacillus subtilis TD7

Q2 Agricultural and Biological Sciences
Bozhong Mu, Fang Liu, Yi-Fan Liu, Yishu Qiao, Yu-Zhe Guo, Fang-Yue Kuang, Xiuying Lin, Jiang Ye, Jin-Feng Liu, Shi‐Zhong Yang, Huizhan Zhang, W. Sand
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引用次数: 4

Abstract

Surfactin is one of the most representative biosurfactants and exhibits excellent surface activity plus other biological effects. It has potential applications in microbial enhanced oil recovery, environmental bioremediation, agricultural bio-control, pharmacy, cosmetics and food industries. The low yield of the surfactant from wild strains is a key restriction for industrial applications. The construction of genetically engineered bacteria by promoter substitution is an effective method to enhance surfactin production, as the promoter is a key element in gene expression. This study focuses on constructing strains with efficient surfactin production by replacing the native srfA promoter by strong promoters. In this study, two different promoter patterns with different homology arm positions were used for srfA promoter substitution. The most efficient installation way was identified as the sequence between the transcriptions start site and ribosome binding site of srfA. Moreover, eight endogenous strong auto-inducible phase-dependent promoters were chosen to substitute the native promoter of srfA using an effective substitution by the CRISPR-Cas9 system. As a result, high surfactin yielding strains with potential application in industry were constructed. According to the results, three constructed strains with promoters P43, PspoVG, and PyvyD showed increased yields of 3.5, 2.8, and 2.3 times over the wild stain B. subtilis TD7.
通过替代枯草芽孢杆菌TD7启动子提高表面素产量
表面活性素是最具代表性的生物表面活性剂之一,具有优异的表面活性和其他生物效应。它在微生物采油、环境生物修复、农业生物控制、制药、化妆品和食品工业中具有潜在的应用前景。野生菌株表面活性剂的低产量是工业应用的一个关键限制。启动子取代构建基因工程菌是提高表面活性素产量的有效方法,因为启动子是基因表达的关键元件。本研究的重点是通过用强启动子取代天然srfA启动子来构建具有高效表面活性素生产的菌株。在本研究中,两种具有不同同源臂位置的不同启动子模式被用于srfA启动子取代。最有效的安装方式是srfA转录起始位点和核糖体结合位点之间的序列。此外,通过CRISPR-Cas9系统的有效取代,选择了8个内源性强自诱导相依赖启动子来取代srfA的天然启动子。从而构建了具有潜在工业应用前景的高表面活性素高产菌株。结果表明,三个启动子分别为P43、PspoVG和PyvyD的构建菌株的产量分别是野生菌株枯草芽孢杆菌TD7的3.5、2.8和2.3倍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Applied Environmental Biotechnology
Applied Environmental Biotechnology Agricultural and Biological Sciences-Agricultural and Biological Sciences (miscellaneous)
CiteScore
3.70
自引率
0.00%
发文量
2
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