酶定向进化的研究策略

定点突变是酶工程中研究酶的结构与功能的常规手段,并广泛用于改变酶的性能.酶的体外定向进化是改造生物催化剂的一种有效的新策略,主要是模拟自然进化进程,通过容错PCR等技术对编码酶的基因进行随机突变,再由DNA改组、交错延伸过程、随机引导重组、递增截短技术等进行突变基因的体外重组,最终经筛选获得所需的酶.本文综述了酶定向进化的研究与应用.

Strategies for Study on Directed Evolution of Enzyme

Site-directed mutagenesis has become the conventional means to study the structure and function ofenzymes. It has been widely used to change the characteristics of enzyme. In vitro molecular directed evolution of enzymeis a new strategy in the engineering of novel biocatalysts, which mainly mimics the process of natural evolution to generaterandom mutation in the genes coding for useful enzymes by the techniques of error-prone PCR, and then in vitro recombinespositive mutations through various methods such as DNA shuffling, staggered extension process, random-primingrecombination, and incremental truncation. Finally, the desired enzymes may be obtained by selection and screening. Recentadvances and applications in the directed evolution of enzyme are reviewed.