The α-mating factor secretion signals and endogenous signal peptides for recombinant protein secretion in Komagataella phaffii

Author:

Zou Chenwei,Lu Lingfang,Wang Shengyan,Zhang Chenshan,Chen Xuequn,Lin Yao,Huang Yide

Abstract

Abstract Background The budding yeast Komagataella phaffii (Pichia pastoris) is widely employed to secrete proteins of academic and industrial interest. For secretory proteins, signal peptides are the sorting signal to direct proteins from cytosol to extracellular matrix, and their secretion efficiency directly impacts the yields of the targeted proteins in fermentation broth. Although the α-mating factor (MF) secretion signal from S. cerevisiae, the most common and widely used signal sequence for protein secretion, works in most cases, limitation exists as some proteins cannot be secreted efficiently. As the optimal choice of secretion signals is often protein specific, more secretion signals need to be developed to augment protein expression levels in K. phaffii. Results In this study, the secretion efficiency of 40 α-MF secretion signals from various yeast species and 32 endogenous signal peptides from K. phaffii were investigated using enhanced green fluorescent protein (EGFP) as the model protein. All of the evaluated α-MF secretion signals successfully directed EGFP secretion except for the secretion signals of the yeast D. hansenii CBS767 and H. opuntiae. The secretion efficiency of α-MF secretion signal from Wickerhamomyces ciferrii was higher than that from S. cerevisiae. 24 out of 32 endogenous signal peptides successfully mediated EGFP secretion. The signal peptides of chr3_1145 and FragB_0048 had similar efficiency to S. cerevisiae α-MF secretion signal for EGFP secretion and expression. Conclusions The screened α-MF secretion signals and endogenous signal peptides in this study confer an abundance of signal peptide selection for efficient secretion and expression of heterologous proteins in K. phaffii.

Funder

Fujian Provincial Science Fund for Distinguished Young Scholar

Fujian provincial health technology project

Publisher

Springer Science and Business Media LLC

Subject

Management, Monitoring, Policy and Law,Energy (miscellaneous),Applied Microbiology and Biotechnology,Renewable Energy, Sustainability and the Environment,Biotechnology

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