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编辑srfA操作子的多个活性位点的腺化域,以构建多样化的表面因子脂

Zhaoyang Wang¹, Li Liu¹, Jiaxun Wei¹

¹School of Chemistry and Chemical Engineering, Tianjin University of Technology, Tianjin, PR China.

Communications biology

|January 6, 2026

概括

研究人员使用CRISPR-Cas9基因编辑设计了表面素变体. 修改后的表面素分子对特定植物病原体具有增强的抗真菌活性,扩大了非核糖体合成酶的潜力.

科学领域:

生物化学生物化学
分子生物学分子生物学
合成生物学合成生物学

背景情况:

表面素是一种由Bacillus物种产生的脂,具有多种生物活性.
非核糖体合成酶 (NRPS),和表面因子合成酶 (SrfA) 一样,是负责脂生物合成的复杂酶.
工程NRPS腺化域对于生成具有改进功能的新型表面素变体至关重要.

研究的目的:

开发一种精确和有效的方法来设计Bacillus pumilus中的srfA操作子.
产生具有改变结构和增强生物活性的新型表面素变体.
为了研究工程表面素变体的抗真菌特性.

主要方法:

使用非特异导向RNA (UgRNA) 和Cas9系统进行srfA操作子的多位点编辑.
在SrfA腺化域中准保存的基因,以引入特定的氨基酸替代.
产生和特征两个工程菌株,WA1和WA2,产生不同的表面素变体:[Ser2]表面素和[Lys3,6]表面素.

主要成果:

成功生成了两种经过工程改造的Bacillus pumilus菌株,具有修改的表面活性蛋白结构.
WA1产生了具有针对Ustilaginoidea virens的强有力的活性[Ser2]表面素.
WA2产生了[lys3,6]surfactin,证明了对Pyricularia oryzae和U. virens的抑制作用.
母菌株对Rhizoctonia solani的有效性有限.

结论:

乌格RNA/Cas9系统能够高效多样化编辑NRPS腺化域.
与野生类型相比,工程化表面素变体表现出明显的和增强的抗真菌活性.
这种方法具有显著的潜力,可以扩大脂的结构和功能多样性,用于各种应用.

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