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在Cupriavidus necator H16中用于全细胞生物催化剂的酶表达

Matteo Vajente¹, Mattia Ghirardi¹, Sandy Schmidt¹

¹Department of Chemical and Pharmaceutical Biology, Groningen Research Institute of Pharmacy, University of Groningen, Groningen, The Netherlands.

Methods in enzymology

|April 27, 2025

概括

复杂分子的可持续合成至关重要. 本研究提出了一种精简的方法,用于设计Cupriavidus necator H16用于体内生物催化,从而实现高效的辅因子再生和可持续的化学生产.

科学领域:

生物技术是生物技术.
合成生物学合成生物学
可持续化学可持续化学

背景情况:

生物催化提供可持续的合成途径,但通常需要辅助因子再生.
在体内生物催化利用微生物代谢进行辅因子再生,但菌株工程可能耗时.
库普里亚维杜斯 (Cupriavidus necator H16) 是一个有前途的微生物底盘,具有光自营代谢和O2耐受性可溶性酶.

研究的目的:

提出一个快速和精简的策略,用于设计Cupriavidus necator H16用于体内生物催化剂.
为了证明一种新的表达等离子体对C. necator H16.16中高效蛋白质表达的实用性.
促进未来使用C. necator H16.16进行体内生物催化工艺的开发.

主要方法:

开发了C. necator H16的基因工程管道,从等离子体克隆开始.
利用最近开发的表达等离子体来有效地传递基因和蛋白质表达.
工程设计的C. necator H16用于表达生物催化应用的模型氧化还原酶.

主要成果:

为C. necator H16.16建立了一个快速和精简的应变工程管道.
在工程化C. necator H16.16中成功证明了模型氧化还原酶的蛋白质表达.
开发的管道大大减少了C. necator H16遗传修饰所需的时间和精力.

结论:

提出的策略使得C. necator H16的高效和快速工程能够用于体内生物催化剂.
这种方法克服了与耗时的基因工程相关的先前限制.
促进使用C. necator H16.16的新型可持续生物催化工艺的发现和开发.

关键词:

自营型细菌是一种自营型细菌.生物催化剂是一种生物催化剂.辅助因子回收利用的回收利用这就是Cupriavidus necator.氧化降解酶是一种氧化降解酶.全细胞生物转化过程

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