蛋白质定向进化及其在微生物代谢调控中的应用

Directed Protein Evolution and Its Applications to Metabolic Regulation in Microbes

蛋白质定向进化是非理性改造蛋白质的一种有效方法。利用蛋白质定向进化技术可以改变代谢流,扩展或构建新的代谢途径,弱化或消除不必要或有害的代谢途径,从而达到提高某种代谢产物产率或降解有害物质的目的。蛋白质定向进化技术在代谢调控中的应用有效拓宽了代谢工程的应用范围。本文介绍了主要的蛋白质定向进化技术如易错PCR技术、DNA改组技术、交错延伸技术和临时模板随机嵌合技术等,评述了蛋白质定向进化技术对微生物细胞代谢中的关键蛋白进行定向改造,从而改善其代谢能力,调控微生物代谢等的应用。

Directed protein evolution is an effective and powerful irrational approach for protein engineering.The use of directed protein evolution technology can change the protein metabolic flow,expand or build a new meta-bolic pathway as well as weaken or eliminate unnecessary or harmful metabolites so as to meet the purposes of increasing the yield of metabolites or degrading harmful substances.The technology of directed protein evolution applied in the metabolic regulations would effectively broaden the ranges of metabolic engineering applications.In this review we introduced the main methods of directed protein evolution such as error-prone PCR,DNA shuffling,Stagger Extension Process （StEP）,Random Chimeragenesis on Transient Templates （RACHITT） and so on.And also we reviewed the applications of directed protein evolution in metabolic regulation to modify the key proteins for improving its metabolic ability and metabolite properties.