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一个双重的CRISPR-Cas/Cre-loxP基因组工程策略,用于稳定的尿酶表达在食品级益生菌中

Xiaoyuan Tang¹, Dianwen Ju¹, Haifeng Hu²

¹School of Pharmaceutical Sciences, Shanghai Engineering Research Center of Immunotherapeutics, Fudan University, Shanghai 201203, PR China.

ACS synthetic biology

|December 23, 2025

概括

我们使用CRISPR-Cas9和Cre/loxP系统设计了一个强大的食品级微生物底盘,Lactococcus lactis NZ9000. 这种增强菌株有效降解酸盐,为益生菌和生物疗法提供稳定的平台.

科学领域:

合成生物学合成生物学
微生物工程微生物工程
益生菌益生菌益生菌

背景情况:

开发具有特定代谢功能的食品级微生物底盘对于健康和营养应用至关重要.
乳球菌 (Lactococcus lactis) 是一种广泛使用的食品级细菌,具有益生菌发展的潜力.

研究的目的:

为增强异质基因表达设计一个强大且基因稳定的Lactococcus lactis底盘.
创建一个具有量身定制的代谢活性的食品级益生菌平台,专门用于尿酸降解.

主要方法:

采用了双基因组工程策略,结合了CRISPR-Cas9敲进和Cre/loxP驱动的基因组减少.
乳球菌乳酸菌NZ9000的基因组被简化,使其能够稳定地表达一种高活性尿酶变体.

主要成果:

工程菌株NZ9000::UA^T-ΔD6在体外表现出增强的尿酶活性 (2.34 U/mL).
大约500μM尿酸的完全降解在20小时内实现.
由此产生的底盘在小鼠中表现出遗传稳定性,生物安全性质和适度的肠道殖民能力.

结论:

双重的CRISPR-Cas和Cre/loxP系统为设计食品级益生菌和活生物疗法提供了一个多功能平台.
这种改造的Lactococcus lactis菌株作为目标代谢活动的稳定底盘,在健康和营养方面推进了合成生物学.

关键词:

这就是CRISPR/Cas9的作用.克雷洛克斯 (Cre/loxPP) 是一个植物.基因编辑基因编辑超血症是什么意思乳酸细菌是一种乳酸细菌.益生菌是一种益生菌.

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