酸酶辅因子生物合成使用在Saccharomyces cerevisiae的线粒体中产生的蛋白质
Katarzyna Dobrzyńska1,2, Ana Pérez-González3, Carlos Echavarri-Erasun1,2
1Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA/CSIC), Pozuelo de Alarcón, Spain.
mBio
|December 21, 2023
概括
研究人员设计了酵母Saccharomyces cerevisiae,以产生生物固化的必需成分. 这一突破涉及7Fe-9S-C-Mo-R-homocitrate]-cofactor (FeMo-co) 组件,可能导致固定谷物并减少对肥料的依赖.
科学领域:
- 生物化学和分子生物学
- 微生物学 微生物学
- 合成生物学 合成生物学
背景情况:
- 生物固化将大气中的N2转化为氨 (NH3),这一过程对生命至关重要,但仅限于二氧化微生物.
- 基酶催化这种转化,其中7Fe-9S-C-Mo-R-同位素]-辅因子 (FeMo-co) 是Mo-依赖酶中的活性位点.
- 工程真核生物体的固需要成功的FeMo-co组装,一个复杂的过程涉及脚手架蛋白质,如NifEN.
研究的目的:
- 确定能够支持 FeMo-co 组装的功能性 NifEN 蛋白质变体在真核模型生物 Saccharomyces cerevisiae 中.
- 使用异构成分在S. cerevisiae线粒体内建立一个完整的FeMo-co生物合成途径.
- 探索结合跨物种化酶组件组件组合的潜力,以优化真核生物中的固定.
主要方法:
- 来自各种类阳菌的nifEN基因库在S. cerevisiae中得到表达,并向线粒体.
- 鉴定了可溶性NifEN蛋白质复合物,其支持FeMo-co形成的能力在体外和在二氧化A. vinelandii中进行了测试.
- FeMo-co生物合成途径在体外复制,使用NifEN,NifB和NifH蛋白质从S. cerevisiae线粒体中表达和净化.
主要成果:
- 在许多NifEN变体中,只有三种在S. cerevisiae线粒体中以可溶的形式积累.
- 其中两个可溶性NifEN变体在体外FeMo-co合成试验中表现出活性.
- 在体外成功建立了一个功能性的FeMo-co生物合成途径,使用来自不同物种的成分,所有这些都是S. cerevisiae产生的.
结论:
- 功能性NifEN蛋白质复合体可以在真核酵母S. cerevisiae的线粒体中被识别并产生.
- 结合多种跨物种的化酶FeMo-co组件组件是实现在真核生物中固化的可行策略.
- 这项工作代表了在真核生物体 (如谷物) 中设计固定能力的重要一步,可能减少对合成肥料的依赖.
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