Engineering of the unfolded protein response pathway in Pichia pastoris: enhancing production of secreted recombinant proteins

Autor:	Astrid Weninger, Zdeněk Knejzlík, Hana Raschmanová, Karin Kovar, Karel Melzoch
Rok vydání:	2021
Předmět:	Unfolded protein response (UPR) Chaperone Protein degradation Endoplasmic-reticulum-associated protein degradation Applied Microbiology and Biotechnology Pichia Pichia pastoris Fungal Proteins Secretion Folding and secretion biology Endoplasmic reticulum Productivity of recombinant protein production General Medicine Mini-Review biology.organism_classification Recombinant Proteins Cell biology Chaperone (protein) Saccharomycetales Unfolded Protein Response biology.protein Unfolded protein response Protein folding Co-expression strategy Biotechnology
Zdroj:	Applied Microbiology and Biotechnology
ISSN:	1432-0614 0175-7598
DOI:	10.1007/s00253-021-11336-5
Popis:	Abstract Folding and processing of proteins in the endoplasmic reticulum (ER) are major impediments in the production and secretion of proteins from Pichia pastoris (Komagataella sp.). Overexpression of recombinant genes can overwhelm the innate secretory machinery of the P. pastoris cell, and incorrectly folded proteins may accumulate inside the ER. To restore proper protein folding, the cell naturally triggers an unfolded protein response (UPR) pathway, which upregulates the expression of genes coding for chaperones and other folding-assisting proteins (e.g., Kar2p, Pdi1, Ero1p) via the transcription activator Hac1p. Unfolded/misfolded proteins that cannot be repaired are degraded via the ER-associated degradation (ERAD) pathway, which decreases productivity. Co-expression of selected UPR genes, along with the recombinant gene of interest, is a common approach to enhance the production of properly folded, secreted proteins. Such an approach, however, is not always successful and sometimes, protein productivity decreases because of an unbalanced UPR. This review summarizes successful chaperone co-expression strategies in P. pastoris that are specifically related to overproduction of foreign proteins and the UPR. In addition, it illustrates possible negative effects on the cell’s physiology and productivity resulting from genetic engineering of the UPR pathway. We have focused on Pichia’s potential for commercial production of valuable proteins and we aim to optimize molecular designs so that production strains can be tailored to suit a specific heterologous product. Key points • Chaperones co-expressed with recombinant genes affect productivity in P. pastoris. • Enhanced UPR may impair strain physiology and promote protein degradation. • Gene copy number of the target gene and the chaperone determine the secretion rate.
Databáze:	OpenAIRE
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