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由有机体依赖的多蛋白质设计使得固化组件能够在线粒体上得到体质表达,这些固化组件的目标是线粒体

Jianguo Yang¹, Nan Xiang¹, Yiheng Liu¹

¹State Key Laboratory of Protein and Plant Gene Research, School of Advanced Agricultural Sciences and School of Life Sciences, Peking University, Beijing 100871, China.

Proceedings of the National Academy of Sciences of the United States of America

|August 16, 2023

概括

在真核生物中,通过将组件向有机细胞来实现工程固 (nif) 基因. 一个新的多蛋白系统使用内源性酵母线粒体蛋白酶进行稳定表达,使线粒体中酶活性成为可能.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
植物科学植物科学

背景情况:

在真核生物体中工程固定 (nif) 基因对于开发固定作物至关重要.
在有机体内,Nif成分的稳定和稳定表达是酶活性的先决条件.
之前使用烟草蚀刻病毒蛋白酶 (TEVp) 的多蛋白系统在酵母中对线粒体向面临挑战.

研究的目的:

开发一种基于多蛋白质的新酶系统,用于稳定的Nif成分在酵母线粒体中的表达.
为了克服TEVp在真核细胞器官的分裂的局限性.
为了使真核细胞中的固定成为可能,并设计线粒体代谢途径.

主要方法:

设计了一个Nif多蛋白系统,用最小的序列取代TEVp裂变部位.
利用基于知识的工程来识别由内源性线粒体处理酶分裂的序列.
在大肠杆菌和随后在酵母线粒体中测试了该系统.

主要成果:

修改后的多蛋白系统在大肠杆菌中实现了与基于TEVp的系统相当的酶活性.
在酵母线粒体中成功实现了Nif成分的稳定和平衡表达.
新系统减少了基因需求,并防止过早的多蛋白质分裂.

结论:

使用内源性线粒体蛋白酶的新型多蛋白系统使酵母线粒体中稳定的Nif成分表达成为可能.
这一策略是有效的工程固定在真核细胞.
这种方法对代谢途径工程具有更广泛的应用,需要线粒体细分.

关键词:

进行线粒体加工的化酶.化酶是一种化酶.这是一种多蛋白质多蛋白质.合成生物学合成生物学

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