巴斯德毕赤酵母的基因改造研究进展  

Recent Advances in Genetic Modification of Pichia Pastoris

作　　者：何旭晖 纪雪梅 HE Xuhui;JI Xuemei(School of Life Science and Technology,China Pharmaceutical University,Nanjing 211198,China)

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京211198

出　　处：《药物生物技术》2024年第6期718-724,共7页Pharmaceutical Biotechnology

摘　　要：巴斯德毕赤酵母是一种甲基营养型酵母,可以利用甲醇、葡萄糖、甘油作为碳源,具有生产成本低、发酵密度高、表达水平较高,副产物少的优势,目前该系统已成功用于胰岛素、胶原蛋白、抗体类药物的生产。然而,作为一种应用于工业生产的表达系统,其具有如蛋白无法分泌表达、表达量低、翻译后修饰不理想、诱导剂甲醇危险不易存放等缺陷。随着基因编辑工具的发展,研究者广泛开展了对巴斯德毕赤酵母基因水平、转录调控和代谢途径的改造,以扩大巴斯德毕赤酵母产品的范围。文章主要介绍了常用的毕赤酵母基因编辑工具研究现状,并简述了基因编辑工具在毕赤酵母糖基化人源化及转录调控改造中的最新应用,最后对毕赤酵母基因编辑研究的发展做出了展望。Pichia Pastoris is methylotrophic yeast that can utilise methanol,glucose,and glycerol as carbon sources,and has the advantages of low production costs,high fermentation density,higher expression levels,and fewer by-products.Currently,this system has been successfully used in the production of insulin,collagen,and antibody drugs.However,as an expression system applied to industrial production,it has defects such as the inability of protein secretion expression,low expression,unsatisfactory post-translational modification,and the inducer methanol is not easy to store.With the development of gene editing tools,researchers have extensively carried out modifications on gene level,transcriptional regulation and metabolic pathways of Pichia Pastoris to expand the scope of product.This paper focuses on the current research on Pichia Pastoris gene editing tools and briefly describes the latest applications of gene editing tools in glycosylation humanisation and transcriptional regulation modification in Pichia Pastoris.Finally,an outlook on the development of gene modification in Pichia Pastoris is given.

关 键 词：巴斯德毕赤酵母 甲基营养型酵母 CRISPR/Cas9 Cre/LoxP 糖基化 底盘细胞 

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