当前位置：
X-MOL 学术
›
J. Am. Chem. Soc.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
High Throughput Screening with SAMDI Mass Spectrometry for Directed Evolution
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-11-11 , DOI: 10.1021/jacs.0c07828 Adam J. Pluchinsky , Daniel J. Wackelin 1 , Xiongyi Huang 1 , Frances H. Arnold 1 , Milan Mrksich
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-11-11 , DOI: 10.1021/jacs.0c07828 Adam J. Pluchinsky , Daniel J. Wackelin 1 , Xiongyi Huang 1 , Frances H. Arnold 1 , Milan Mrksich
Affiliation
|
Advances in directed evolution have led to an exploration of new and important chemical transformations; however, many of these efforts still rely on the use of low-throughput chromatography-based screening methods. We present a high-throughput strategy for screening libraries of enzyme variants for improved activity. Unpurified reaction products are immobilized to a self-assembled monolayer and analyzed by mass spectrometry, allowing for direct evaluation of thousands of variants in under an hour. The method was demonstrated with libraries of randomly mutated cytochrome P411 variants to identify improved catalysts for C-H alkylation. The technique may be tailored to evolve enzymatic activity for a variety of transformations where higher throughput is needed.
中文翻译：
使用 SAMDI 质谱进行定向进化的高通量筛选
定向进化的进步导致了对新的重要化学转化的探索；然而，许多这些努力仍然依赖于使用基于低通量色谱的筛选方法。我们提出了一种高通量策略，用于筛选酶变体文库以提高活性。未纯化的反应产物被固定在自组装单层上，并通过质谱分析，允许在一小时内直接评估数千个变体。该方法用随机突变的细胞色素 P411 变体文库进行了证明，以鉴定用于 CH 烷基化的改进催化剂。该技术可以被定制以针对需要更高通量的各种转化来进化酶活性。
更新日期：2020-11-11
中文翻译：
使用 SAMDI 质谱进行定向进化的高通量筛选
定向进化的进步导致了对新的重要化学转化的探索；然而，许多这些努力仍然依赖于使用基于低通量色谱的筛选方法。我们提出了一种高通量策略，用于筛选酶变体文库以提高活性。未纯化的反应产物被固定在自组装单层上，并通过质谱分析，允许在一小时内直接评估数千个变体。该方法用随机突变的细胞色素 P411 变体文库进行了证明，以鉴定用于 CH 烷基化的改进催化剂。该技术可以被定制以针对需要更高通量的各种转化来进化酶活性。
京公网安备 11010802027423号