新Yarrowia lipolytica底盘菌株用于工业酶生产
Djamila Onésime1, Esteban Lebrun2, Goran Stanajic Petrovic2
1INRAE, AgroParisTech, Micalis Institute, UMR1319, Université Paris-Saclay, Domaine de Vilvert, 78350, Jouy-en-Josas, France. djamila.onesime@inrae.fr.
Microbial cell factories
|July 10, 2025
概括
优化的酵母菌株增强了重组蛋白质 (rProtein) 的生产. 在Yarrowia lipolytica底盘菌株中的广泛的蛋白酶删除改善了r 蛋白质产量而不需要多个基因拷贝.
科学领域:
- 微生物学 微生物学
- 生物技术是生物技术.
- 合成生物学 合成生物学
背景情况:
- Yarrowia lipolytica是一个强大的微生物工厂,用于生产生物分子和重组蛋白质 (rProtein).
- 菌株开发的重点是辅助性标记物,分泌途径优化和可诱导的表达.
- 需要进一步的生理修饰来满足对蛋白质合成的工业需求.
研究的目的:
- 为了进一步优化Yarrowia lipolytica作为rProtein生产的平台.
- 开发具有增强基因工程能力和减少蛋白质分解活性的新底盘菌株.
- 为了评估辅助性,载体元素去除和蛋白酶缺失对rProtein产量的影响.
主要方法:
- 为先进的基因工程引入第三种辅食性.
- 移除细菌载体元素,包括抗生素耐药性基因.
- 广泛删除细胞外蛋白酶和过氧化酶基因,以创建缺乏蛋白酶的菌株.
- 在工程菌株中评估不同基因拷贝数 (一个,两个或三个) 的蛋白质生产效率.
主要成果:
- 新建的底盘菌株 (JMY9438,JMY9451/9452) 具有三重辅性.
- 菌株JMY9451/9452由于五个蛋白酶基因被删除,其蛋白质分解活性显著降低.
- 蛋白质表达增加,基因拷贝数增加到两个;三个拷贝没有进一步改善.
- 蛋白酶被删除的菌株具有单个基因副本,实现了每细胞最高的rProtein;蛋白酶和两个副本的菌株产生了最高的绝对rProtein水平.
结论:
- 一个新一代的Yarrowia lipolytica底盘菌株已被开发用于优化rProtein生产.
- 广泛的蛋白酶删除为高水平的r蛋白质合成创造了高性能遗传背景.
- 有效的r蛋白质生产可以在不依赖多拷贝表达策略的情况下实现.
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