生产菌株的筛选与选择：多形汉逊酵母中分泌蛋白的表达与分析

Screening and Selection of Production Strains: Secretory Protein Expression and Analysis in Hansenula polymorpha.

作者信息

Yoo Su Jin, Moon Hye Yun, Kang Hyun Ah

机构信息

Department of Life Science, College of Natural Science, Chung-Ang University, Seoul, Republic of Korea.

出版信息

Methods Mol Biol. 2019;1923:133-151. doi: 10.1007/978-1-4939-9024-5_5.

DOI:10.1007/978-1-4939-9024-5_5

PMID:30737738

Abstract

The thermotolerant methylotrophic yeast Hansenula polymorpha has been used as a host for the high-level production of recombinant proteins from industrial enzymes to therapeutic proteins. Despite favorable characteristics of the H. polymorpha-based platform for application to heterologous gene expression, several problems and limitations, such as over-glycosylation and proteolytic degradation, can be encountered in the development of production strains for secretory proteins. Here, H. polymorpha genetic tools and host strains, developed for authentic processing and modification of secretory recombinant proteins, are introduced with the analytical protocols.

摘要

嗜热甲基营养型酵母多形汉逊酵母已被用作宿主，用于从工业酶到治疗性蛋白质的重组蛋白的高水平生产。尽管基于多形汉逊酵母的平台在应用于异源基因表达方面具有良好特性，但在分泌蛋白生产菌株的开发中可能会遇到一些问题和限制，例如过度糖基化和蛋白水解降解。本文将介绍用于分泌型重组蛋白的真实加工和修饰的多形汉逊酵母遗传工具和宿主菌株，并给出分析方案。