用SAMDI质谱测定定向进化的高吞吐量选
Adam J Pluchinsky, Daniel J Wackelin1, Xiongyi Huang1
1Division of Chemistry and Chemical Engineering MC 210-41, California Institute of Technology, Pasadena, California 91125, United States.
Journal of the American Chemical Society
|November 11, 2020
概括
研究人员使用质谱测量开发了一种高通量查方法. 这种方法可以快速评估成千上万种酶催化剂以改善化学转化,加速酶的进化.
科学领域:
- 生物催化
- 酶工程
- 分析化学
背景情况:
- 定向进化促进了酶的发现,但往往使用低通量选.
- 基于色谱的方法限制了酶变体的评估规模和速度.
研究的目的:
- 为选酶变体提出一个高通量策略.
- 能够直接评估成千上万种酶变体以提高活性.
主要方法:
- 在自组合单层上固定未净化的反应产物.
- 固定产品的质谱分析.
- 用于随机突变的P411变体库.
主要成果:
- 在选酶变异方面表现出高通量策略.
- 使用P411细胞变异物成功鉴定出改善的C-H化催化剂.
- 在不到一个小时的时间里, 直接评估了成千上万的变体.
结论:
- 开发的方法显著增加了酶查的吞吐量.
- 这种技术可以适应各种化学转换的酶活性.
- 加快新型和改进的生物催化剂的发现.
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