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在Candida viswanthii中开发基于CRISPR的合成生物学工具包,并对应激响应的Ena1样蛋白的功能分析

Xin-Yue Li¹, Kai Li¹, Feng-Li Zhang¹

¹Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240, China.

Synthetic and systems biotechnology

|November 10, 2025

概括

研究人员开发了一种新的CRISPR/Cas9和Cre/loxP基因组编辑系统,用于*Candida viswanathii*. 这种高效的工具可以实现快速,代的基因改造,以改善工业酵母菌株的生物生产.

科学领域:

微生物学微生物学
合成生物学合成生物学
生物技术是生物技术.

背景情况:

工业 *Candida* 种类对于生产酶,生物化学物质和单细胞蛋白质非常有价值.
现有的合成生物学工具用于操纵*Candida*物种是有限的,阻碍了它们的生物技术应用.

研究的目的:

为*Candida viswanathii*建立一个高效的基因组编辑策略.
为了实现增强生物生产的快速,代和多重基因组编辑.

主要方法:

结合CRISPR/Cas9和Cre/loxP系统用于*C. viswanathii*的基因组编辑.
开发了一个快速代编辑周期 (6天内) 的平台.
应用了多重编辑和多重复制基因集成系统 (最多7个副本).

主要成果:

在C.viswanathii*中实现了100%的编辑效率.
功能性特征的g144,一种类似Ena1的蛋白质,揭示了它在pH和Na+应激反应和DDA产生中的作用.
证明了g144的破坏或过度表达会降低 dodecanedioic 酸的产生.

结论:

开发的基因组编辑工具包显著提升了对*Candida*物种的合成生物学工具.
该平台促进了高效的基因改造,以优化可持续生物生产中的工业酵母菌株.
对g144的功能性特征提供了对生物工艺工程相关的 *Candida *生理学的见解.

关键词:

这就是CRISPR/Cas9的作用.在Candida viswanathii中发现的类似Ena1的蛋白质g144 代编辑代编辑多拷贝集成的整合.多重编辑多重编辑.

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