人工金属酶在体内转化过程中的定向演化
Markus Jeschek1, Raphael Reuter2, Tillmann Heinisch2
1Department of Biosystems Science and Engineering, ETH Zurich, Basel CH-4058, Switzerland.
Nature
|August 30, 2016
概括
研究人员在大肠杆菌周等离子体内设计了一种人工金属酶,用于olefin转化,使得体内进化和扩展生物催化用于非自然反应.
科学领域:
- 生物催化和合成生物学
- 蛋白质工程和定向进化研究
- 有机金属化学 有机金属化学
背景情况:
- 生物催化剂传统上使用天然酶,但定向进化和人工金属酶扩大了新反应的能力.
- 人工金属酶将过渡金属与酶支架相结合,实现多功能,工程化催化.
- 现有的人工金属酶方法通常需要蛋白质净化,限制了体内应用.
研究的目的:
- 开发一种用于非生物反应的人工金属酶体体内进化的方法.
- 为了设计一个人造的金属酶来进行olefin转化,这种反应在自然界中不存在.
- 克服蛋白质净化的局限性,用于人工金属酶的开发和应用.
主要方法:
- 在大肠杆菌 (Escherichia coli) 的周周等离子体内对人工金属酶进行细分.
- 人工金属酶的定向进化,以增强体内活性和稳定性.
- 利用周等离子体作为反应来缓解像谷氨这样的细胞成分的抑制.
主要成果:
- 在体内成功组装和演化一种功能性人工金属酶 ("甲酶") 用于烯酸甲酶.
- 与使用纯化蛋白质的传统方法相比,实现了针对性进化的显著更高的吞吐量.
- 进化的转基因酶与商业催化剂的比较良好,并与各种基质的活性有很好的比较.
结论:
- 周等离子体分离策略使人工金属酶在无生物反应中能够在体内高效地进化.
- 这种方法扩大了人工金属酶在非自然代谢和生物催化剂中的应用潜力.
- 开发的工作流提供了一个强大的平台,用于进一步设计用于各种催化功能的人工金属酶.
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