毕赤酵母高效表达外源蛋白的分子水平策略  

Molecular level strategy for high expression of foreign protein in Pichia pastoris

作　　者：张欣然 凌焱 杨英 ZHANG Xinran;LING Yan;YANG Ying(Institute of Radiation Medicine,Academy of Military Medical Science,Beijing 100850,China;School of Life Sciences,Hebei University,Baoding 071000,China)

机构地区：[1]军事医学研究院辐射医学研究所,北京100850 [2]河北大学生命科学学院,河北保定071000

出　　处：《食品与发酵工业》2022年第17期321-328,共8页Food and Fermentation Industries

基　　金：国家科技重大专项(2018ZX10101003);中国工程院咨询研究项目(2020-XY-61-03)。

摘　　要：毕赤酵母表达系统具有许多优异特性,并且是目前最成功的外源蛋白表达系统之一。通过近几十年的快速发展,该系统已引起各界的广泛关注,并产生了巨大的经济和社会价值。与其他现有表达系统相比,毕赤酵母在目的蛋白翻译后修饰上有着极大优势,已被广泛应用于表达外源蛋白。但是另一方面,由于该系统受包括外源基因的特性、菌种、表达环境和发酵技术等多种因素的影响,,其在不同外源蛋白的表达上也表现出很大差异。该文着眼于分子水平上的基因转录、蛋白质加工、蛋白质降解以及转运分泌4个模块,介绍了利用毕赤酵母表达系统通过优化密码子、高拷贝数外源基因、引导肽筛选、敲除蛋白酶基因、共表达促折叠因子等途径提高外源蛋白表达效率的相关策略,并简要展望了该系统的发展前景,以期为构建高效毕赤酵母细胞工厂提供指导。Pichia pastoris expression system has many excellent characteristics and is one of the most successful foreign protein expression systems.Through the rapid development in recent decades,it has attracted wide attention from all walks of life,and has produced huge economic and social value.Compared with other existing expression systems,P.pastoris has great advantages in post-translational modification of target proteins,and has been widely used to express exogenous proteins.However,on the other hand,the system is affected by many factors,including the characteristics of exogenous genes,bacterial species,expression environment and fermentation technology,so it also shows great differences in the expression of different exogenous proteins.Gene transcription module,this paper focuses on the molecular level of protein secretion and degradation of modules and transport four modules,and introduces the use of P.pastoris expression system by optimizing codon,high copy number exogenous gene screening,gene knockout protease,and guide the peptides were folded factors to improve the efficiency of heterologous protein expression of related policies,The development prospect of the system is briefly prospected.

关 键 词：巴斯德毕赤酵母 酵母表达系统 高效表达 分子水平 外源蛋白 

分 类 号：Q78[生物学—分子生物学] TQ920.1[轻工技术与工程—发酵工程]