毕赤酵母外源蛋白分泌及折叠途径的改良进展

doi:10.13304/j.nykjdb.2023.0160

摘要/Abstract

摘要：

工业、农业和医药等领域常用的活性蛋白和工业酶大多数通过异源表达系统获得。毕赤酵母（Pichia pastoris）是优秀的外源蛋白表达宿主之一，以毕赤酵母为宿主的表达系统具有遗传稳定性好、翻译后修饰、蛋白表达和分泌水平高及生产成本低等优点，但在高效表达过程中外源蛋白过量聚集会导致目标蛋白不能正确折叠和有效分泌，从而影响蛋白表达水平。概述了通过信号肽优化、分子伴侣优化以及融合蛋白表达等分泌及折叠途径的改良，从而促进外源蛋白高效表达的研究进展。

关键词: 毕赤酵母, 分泌折叠途径, 分泌信号肽, 分子伴侣, 融合表达

Abstract:

Most of the active proteins and industrial enzymes commonly used in industry， agriculture and medicine are obtained through heterologous expression systems. Pichia pastoris is one of the excellent hosts for expressing heterologous proteins. This expression system has the advantages including good genetic stability， high protein expression and secretory level， post-translational modification system， and low cost etc. However， excessive aggregation of heterologous proteins usually leads to incorrect fold and ineffective secretion of the target proteins， thus decling protein expression level. This paper reviewed the progress in the study on exogenous protein expression in Pichia pastoris by engineering the secretion and folding pathways such as signal peptide optimization， chaperone optimization and fusion protein expression.

Key words: Pichia pastoris, secretion and folding pathways, signal peptide, molecular chaperones, fusion expression

中图分类号:

S182

路泽群, 刘宁, 张红莲, 王苑, 黄火清. 毕赤酵母外源蛋白分泌及折叠途径的改良进展[J]. 中国农业科技导报, 2024, 26(1): 18-27.

Zequn LU, Ning LIU, Honglian ZHANG, Yuan WANG, Huoqing HUANG. Improvement of Heterologous Protein Secretion and Folding Pathways of Pichia pastoris[J]. Journal of Agricultural Science and Technology, 2024, 26(1): 18-27.

图/表 3

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基因 Gene	融合标签 Fusion tag	连接肽 Linker	融合蛋白产量 Fusion protein production	参考文献 Reference
hFGF21	HSA	柔性 Flexibility	0.94 mg·mL^-1	［68］
rhIL-2	HSA	刚性 Rigidity	32 mg·L^-1	［75］
cIFN	HSA	柔性 Flexibility	86 mg·L^-1	［67］
AMP	HSA	His-TEV	700 mg·L^-1	［76］
SpG	辣根过氧化物酶 Horseradish peroxidase（HRP）	柔性 Flexibility	113 mg·L^-1	［77］
hBMP2	人样胶原蛋白 Human-like collagen（HLC）	柔性 Flexibility	1 g·L^-1	［78］
LfcinB–hLY	抗氧化肽 Antioxidant peptide	柔性 Flexibility	3.6 mg·L^-1	［79］
rLG	SUMO	柔性 Flexibility	4.32 mg·L^-1	［55］
NZ17074	SUMO3	甲酸切割位点 Formic acid cleavage site	4.1 mg·L^-1	［56］
ANL	SUMO	刚性 Rigidity	1.82 mg·L^-1	［57］
GrB	MBP	弗林蛋白酶Furin	ND	［60］
SS	木聚糖酶 Xylanase	柔性 Flexibility	3.640 g·L^-1	［64］
HEWL	木聚糖酶 Xylanase	Kex2、Ste13	HEWL产量3.5 g·L^-1 HEWL yield 3.5 g·L^-1	［65］

基因 Gene	融合标签 Fusion tag	连接肽 Linker	融合蛋白产量 Fusion protein production	参考文献 Reference
hFGF21	HSA	柔性 Flexibility	0.94 mg·mL^-1	［68］
rhIL-2	HSA	刚性 Rigidity	32 mg·L^-1	［75］
cIFN	HSA	柔性 Flexibility	86 mg·L^-1	［67］
AMP	HSA	His-TEV	700 mg·L^-1	［76］
SpG	辣根过氧化物酶 Horseradish peroxidase（HRP）	柔性 Flexibility	113 mg·L^-1	［77］
hBMP2	人样胶原蛋白 Human-like collagen（HLC）	柔性 Flexibility	1 g·L^-1	［78］
LfcinB–hLY	抗氧化肽 Antioxidant peptide	柔性 Flexibility	3.6 mg·L^-1	［79］
rLG	SUMO	柔性 Flexibility	4.32 mg·L^-1	［55］
NZ17074	SUMO3	甲酸切割位点 Formic acid cleavage site	4.1 mg·L^-1	［56］
ANL	SUMO	刚性 Rigidity	1.82 mg·L^-1	［57］
GrB	MBP	弗林蛋白酶Furin	ND	［60］
SS	木聚糖酶 Xylanase	柔性 Flexibility	3.640 g·L^-1	［64］
HEWL	木聚糖酶 Xylanase	Kex2、Ste13	HEWL产量3.5 g·L^-1 HEWL yield 3.5 g·L^-1	［65］

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