酶祖先序列重建与定向进化  

Enzyme ancestral sequence reconstruction and directed evolution

作　　者：张锟 戴翊飞 孙金娣 陆佳晨 陈可泉[1] Kun Zhang;Yifei Dai;Jindi Sun;Jiachen Lu;Kequan Chen(College of Biotechnology and Pharmaceutical Engineering,Nanjing Tech University,Nanjing 211816,Jiangsu,China)

机构地区：[1]南京工业大学生物与制药工程学院,江苏南京211816

出　　处：《生物工程学报》2021年第12期4187-4200,共14页Chinese Journal of Biotechnology

基　　金：国家重点研发计划(No.2018YFA0901500);国家自然科学基金(No.21908099);中国博士后面上项目(No.2020M681596)资助。

摘　　要：酶祖先序列重建是指通过计算机算法推导来自灭绝生物的祖先酶的氨基酸序列的技术。通常可分为6个步骤,依次为现代酶的核酸/氨基酸序列收集、多序列比对、系统发育树构建、祖先酶序列的计算机推测、基因克隆、酶学性质表征。该方法广泛应用于研究分子在行星时间尺度上对环境条件不断变化的适应性和进化机制。随着酶在生物催化领域中扮演越来越重要的角色,该方法逐渐成为研究酶序列、结构和功能关系的有力手段。同时,祖先酶大多具有温度稳定性、突变稳定性等特性,使其成为进一步定向进化的理想蛋白质支架。文中综述了酶祖先序列重建的计算机算法、应用和常用计算机软件,并结合最新研究进展,展望其在酶定向进化领域中的应用前景。The amino acid sequence of ancestral enzymes from extinct organisms can be deduced through in silico approach termed ancestral sequence reconstruction(ASR).ASR usually has six steps,which are the collection of nucleic acid/amino acid sequences of modern enzymes,multiple sequence alignment,phylogenetic tree construction,computational deduction of ancestral enzyme sequence,gene cloning,and characterization of enzyme properties.This method is widely used to study the adaptation and evolution mechanism of molecules to the changing environmental conditions on planetary time scale.As enzymes play key roles in biocatalysis,this method has become a powerful method for studying the relationship among the sequence,structure,and function of enzymes.Notably,most of the ancestral enzymes show better temperature stability and mutation stability,making them ideal protein scaffolds for further directed evolution.This article summarizes the computer algorithms,applications,and commonly used computer software of ASR,and discusses the potential application in directed evolution of enzymes.

关 键 词：祖先序列重建 定向进化 祖先酶 酶结构与功能关系 生物催化 

分 类 号：Q814.2[生物学—生物工程]