探索林烯合成酶的自然多样性和涉及Escherichia coli中基质特异性的分子决定因素
Clement P M Scipion1,2, Jérémy Esque3, Shreyash Borkar2
1CNRS@CREATE, 1 Create Way, #08-01 Create Tower, 138602 Singapore.
Journal of agricultural and food chemistry
|May 21, 2025
概括
优化烯合成酶 (LS) 是微生物生产这种有价值的单烯的关键. 来自Agastache rugosa的工程LS显著提高了烯的产量,提高了其在各种行业中的使用.
科学领域:
- 生物技术是生物技术.
- 代谢工程是代谢工程.
- 酵素工程是什么意思 酵素工程
背景情况:
- 利蒙是一种高需求的性单烯,具有广泛的工业应用.
- 微生物生物合成氨酸通常受到低效的氨酸合成酶 (LS) 活动的限制.
- 优化LS对于增强微生物细胞工厂中的利蒙生产至关重要.
研究的目的:
- 为了识别和表征新的烯合成酶 (LS),以改善烯生物合成.
- 为增强活动和改变基质特异性设计选定的LS.
- 增加从可再生资源中生物生产的利蒙反体.
主要方法:
- 序列数据挖掘以识别十个LS同类.
- 在微生物宿主中对LS同类的体内测试,这些宿主积累了geranyl pyrophosphate (GPP) 或neryl pyrophosphate (NPP).
- 在基分析和局部定向突变发生的工程师LS,专注于基质的特异性和活性.
主要成果:
- 来自Agastache rugosa的一种利蒙合成酶在体内显示了对NPP而不是GPP的偏好.
- 工程改造的Agastache rugosa LS突变体表现出改变的基质特异性.
- 一种特定的突变组合 (S8K/I265V/E276P/P277R/A281K/N282T/I285Q/I286L) 导致GPP的烯产量增加了4.8倍,NPP的烯产量增加了1.9倍.
- 该工程酶主要产生 (+) - 烯,在使用NPP时大约有85-90%的反体过剩.
结论:
- 氨酸合成酶的工程可以显著提高微生物系统中的氨酸产量.
- 已识别和设计的LS变体为生产利蒙反体提供了改进的途径.
- 这项工作支持从可持续的碳来源提升有价值的单二烯的生物生产.
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