Directed evolution strategies for improved enzymatic performance.
Directed evolution strategies for improved enzymatic performance.
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DOI：
10.1186/1475-2859-4-29
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发表时间：
2005-10-07
影响因子：
6.4
通讯作者：
Dalby, PA
中科院分区：
文献类型：
--
作者：
Hibbert, EG;Dalby, PA
利用定向进化技术模拟实验室时间尺度上的进化，利用改变酶的活性、特异性和稳定性的工程技术，现在已经得到了很好的确立。然而，要普遍接受这些方法作为获得用于有机合成的新生物催化剂的途径，需要进一步改进这些方法，使其易于使用，并获得比目前可能的更显著的酶性质变化。文库设计和随机突变方法的最新进展，与新的筛选和选择工具相结合，继续推动定向进化的潜力。例如，蛋白质工程师现在开始将对蛋白质结构和功能的大量知识和理解应用到集中定向进化库的设计中，与以前喜欢的整个基因的随机突变和重组相比，结果令人震惊。在模拟文库筛选实验过程的计算设计技术方面取得的重大进展，现在也使人们能够在更大的序列空间区域搜索那些被广泛理解并因此可能建模的催化反应。有机合成的生物催化除了使用分离的酶外，还经常使用全细胞，要么用于单一反应，要么用于通过整个代谢途径的转化。由于新陈代谢工程正在开发许多新的全细胞生物催化剂，定向进化改善这些初始设计的潜力也开始实现。
The engineering of enzymes with altered activity, specificity and stability, using directed evolution techniques that mimic evolution on a laboratory timescale, is now well established. However, the general acceptance of these methods as a route to new biocatalysts for organic synthesis requires further improvement of the methods for both ease-of-use and also for obtaining more significant changes in enzyme properties than is currently possible. Recent advances in library design, and methods of random mutagenesis, combined with new screening and selection tools, continue to push forward the potential of directed evolution. For example, protein engineers are now beginning to apply the vast body of knowledge and understanding of protein structure and function, to the design of focussed directed evolution libraries, with striking results compared to the previously favoured random mutagenesis and recombination of entire genes. Significant progress in computational design techniques which mimic the experimental process of library screening is also now enabling searches of much greater regions of sequence-space for those catalytic reactions that are broadly understood and, therefore, possible to model. Biocatalysis for organic synthesis frequently makes use of whole-cells, in addition to isolated enzymes, either for a single reaction or for transformations via entire metabolic pathways. As many new whole-cell biocatalysts are being developed by metabolic engineering, the potential of directed evolution to improve these initial designs is also beginning to be realised.
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影响因子：
56.9
作者：
Korkegian, A;Black, ME;Stoddard, BL
通讯作者：
Stoddard, BL
影响因子：
15
作者：
Amstutz, P;Pelletier, JN;Plückthun, A
通讯作者：
Plückthun, A
DOI：
10.1073/pnas.212627499
发表时间：
2002-12-10
影响因子：
11.1
作者：
Hayes, RJ;Bentzien, J;Dahiyat, BI
通讯作者：
Dahiyat, BI
DOI：
10.1073/pnas.251555398
发表时间：
2001-12-04
影响因子：
11.1
作者：
Bolon, DN;Mayo, SL
通讯作者：
Mayo, SL
影响因子：
6.4
作者：
Hibbert, EG;Dalby, PA
通讯作者：
Dalby, PA