模拟指导工程使得Selinadiene合成酶的功能转换成为向氧化转化的功能转换
Prabhakar L Srivastava1, Sam T Johns2, Angus Voice2
1School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, U.K.
概括
研究人员设计了一种甲合成酶,以产生一种特定的氧化甲. 这种以模拟为指导的方法实现了48%的产量,证明了创造复杂分子的潜力.
科学领域:
- 生物催化和代谢工程 生物催化和代谢工程
- 合成生物学 合成生物学
- 酶工程是什么? 酶工程是什么?
背景情况:
- 由于复杂的机制和对酶结构功能关系的有限理解,为特定产品设计合成酶是很困难的.
- 甲合成酶 (SdS) 呈现出多样化的循环路径,对向产品形成构成挑战.
研究的目的:
- 为了设计一个氨酸-4(15),7(11) - - 合成酶 (SdS),以产生预定义的化.
- 利用原子模拟和现场定向突变发生来指导酶工程的努力.
- 通过微生物发酵来证明通过微生物发酵生产工程性甲的可行性.
主要方法:
- 采用原子模拟来理解酶-碳酸相互作用,并指导突变发生.
- 利用位点定向的突变发生来产生SdS G305E变种.
- 优化反应条件,包括pH值,以提高产品产量.
- 整合了工程酶和美酸通路基因到BL21(DE3) 细胞中进行发酵.
主要成果:
- 这种SdS G305E变种最初产生了20%的-7(11) -en-4-ol.
- 将pH值优化为6.0增加了单-7-en-4-ol的产量,达到48%.
- 在批量发酵中实现了10 mg/L的林-7(11) -en-4-ol的生产规模.
结论:
- 模拟引导的工程是有效的创建特尔合成酶,产生特定的氧化二二.
- 工程设计的SdS G305E变种显示出生物催化生产复杂基烯的潜力.
- 这种方法为可持续生产有价值的自然产品开辟了道路.
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