Open AccessAvoiding entry into intracellular protein degradation pathways by signal mutations increases protein secretion in Pichia pastoris
Author(s) -
Ito Yoichiro,
Ishigami Misa,
Hashiba Noriko,
Nakamura Yasuyuki,
Terai Goro,
Hasunuma Tomohisa,
Ishii Jun,
Kondo Akihiko
Publication year - 2022
Publication title -
microbial biotechnology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.287
H-Index - 74
ISSN - 1751-7915
DOI - 10.1111/1751-7915.14061
Subject(s) - pichia pastoris , signal peptide , secretory protein , secretion , secretory pathway , saccharomyces cerevisiae , protein targeting , pichia , intracellular , vacuole , biochemistry , biology , protein degradation , protein biosynthesis , yeast , microbiology and biotechnology , amino acid , unfolded protein response , recombinant dna , endoplasmic reticulum , gene , membrane protein , golgi apparatus , cytoplasm , membrane
Summary In our previous study, we serendipitously discovered that protein secretion in the methylotrophic yeast Pichia pastoris is enhanced by a mutation (V50A) in the mating factor alpha (MFα) prepro‐leader signal derived from Saccharomyces cerevisiae . In the present study, we investigated 20 single‐amino‐acid substitutions, including V50A, located within the MFα signal peptide, indicating that V50A and several single mutations alone provided significant increase in production of the secreted proteins. In addition to hydrophobicity index analysis, both an unfolded protein response (UPR) biosensor analysis and a microscopic observation showed a clear difference on the levels of UPR induction and mis‐sorting of secretory protein into vacuoles among the wild‐type and mutated MFα signal peptides. This work demonstrates the importance of avoiding entry of secretory proteins into the intracellular protein degradation pathways, an observation that is expected to contribute to the engineering of strains with increased production of recombinant secreted proteins.