Directed enzyme evolution: beyond the low-hanging fruit
Directed enzyme evolution: beyond the low-hanging fruit
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DOI：
10.1016/j.sbi.2012.03.010
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发表时间：
2012-08-01
影响因子：
6.8
通讯作者：
Tawfik, Dan S.
中科院分区：
文献类型：
--
作者：
Goldsmith, Moshe;Tawfik, Dan S.
定向进化领域已经发展到可以针对非天然底物和反应设计酶，使其具有可与天然酶相媲美的催化效率和区域特异性或立体特异性。在这里，我们描述了促成这一进展的概念和方法的进步。我们提出基于小型文库的方法，该文库富含改进的变体并携带补偿性稳定突变。此类文库可以与低通量筛选相结合，提供高精度并直接靶向所需的底物和反应条件，从而提供高度改进的变体。
The field of directed evolution has progressed to the point where it is feasible to engineer enzymes for unnatural substrates and reactions with catalytic efficiencies and regio-specificity or stereo-specificity that rival those of natural enzymes. Here, we describe the conceptual and methodological advances that have enabled this progress. We address methodologies based on small libraries enriched with improved variants and carrying compensatory stabilizing mutations. Such libraries can be combined with low-throughput screens that provide high accuracy and directly target the desired substrate and reaction conditions, and thereby provide highly improved variants.