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

0543022 - ÚOCHB 2022 RIV DE eng J - Journal Article
Raschmanová, H. - Weninger, A. - Knejzlík, Zdeněk - Melzoch, K. - Kovar, K.
Engineering of the unfolded protein response pathway in Pichia pastoris: enhancing production of secreted recombinant proteins.
Applied Microbiology and Biotechnology. Roč. 105, č. 11 (2021), s. 4397-4414. ISSN 0175-7598. E-ISSN 1432-0614
Institutional support: RVO:61388963
Keywords : Pichia pastoris * productivity of recombinant protein production * folding and secretion * unfolded protein response (UPR) * chaperone * co-expression strategy
OECD category: Biochemistry and molecular biology
Impact factor: 5.560, year: 2021
Method of publishing: Open access
https://doi.org/10.1007/s00253-021-11336-5

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.
Permanent Link: http://hdl.handle.net/11104/0320332