毕赤酵母高效表达外源蛋白的相关策略及研究进展

Related Strategies and Research Progress of Efficient Expression of Heterologous Proteins in Pichia pastoris

蛋白类药物和工业酶的异源表达在各类生物制药产业和工业应用中具有重要意义。巴斯德毕赤酵母(Pichia pastoris)作为最为成功的真核蛋白表达系统之一,近20年来经历飞速发展并受到广泛关注,产生了巨大的经济和社会价值。但另一方面,由于该系统受到包括外源基因本身特性、菌种特性、表达环境以及发酵工艺等在内的诸多因素的影响,其对于不同外源蛋白的表达情况也表现出较大差异。为解决这一难题,研究人员尝试使用了各种策略,如密码子优化、高拷贝菌株筛选、引导肽筛选、敲除蛋白酶基因和甘油运转体基因、共表达促折叠因子和透明颤菌血红蛋白,以及诱导条件、辅助表达物添加和发酵工艺的优化。本文对提高此系统外源蛋白表达量的相关策略及产业化研究进展进行了综述,并简要展望此系统的产业化发展前景。

Heterologous expression of therapeutic proteins and industrial enzymes is of great significance in biopharmaceutical industry and industry applications. As one of the most successful eukaryotic expression systems, Pichia pastoris has drawn extensive attentions. It developed rapidly and brought not only huge commercial value but also enormous social value. But on the other side, the expression level of different proteins by this system was quite different due to several factors, such as the characteristics of heterologous gene, the characteristics of expression strain, the expression environment and the fermentation process. To handle this problem, researchers have tried a variety of strategies including codon optimization, screening for strains with high gene copy number and appropriate signal peptide, disruption of protease gene and glycerol transporter gene, co-expression with chaperones and Vitreosci/la hemoglobin, as well as optimization of the induction condition, additional supplements and the fermentation process. In this paper, such strategies which increase the expression efficiency of the therapeutic proteins and industrial enzymes in Pichia pastoris system are reviewed and the prospects of this system are also briefly elaborated.