The Degree and Length of O-glycosylation of Recombinant Proteins Produced in Pichia Pastoris Depends on the Nature of the Protein and the Process Type.

This study aims to reveal the influence of different cultivation strategies on O-mannosylation profiles in P. pastoris. Sixteen different model proteins, produced by different P. pastoris strains, were analyzed for their O-glycosylation profile. Based on the obtained data, human serum albumin (HSA) was chosen to be produced in fast and slow growth fed batch fermentations by using common promoters, PGAP and PAOX1 . After purification and protein digestion, glycopeptides were analyzed by LC/ESI-MS. In the samples expressed with PGAP it was found that the degree of glycosylation was slightly higher when a slow growth rate was used, regardless of the efficiency of the producing strain. The highest glycosylation intensity was observed in HSA produced with PAOX1 . The results indicate that the O-glycosylation level is markedly higher when the protein is produced in a methanol-based expression system. This article is protected by copyright. All rights reserved.
PMID: 32975831 [PubMed - as supplied by publisher]

Source: Biotechnology Journal - September 24, 2020 Category: Biotechnology Authors: Radoman B, Grünwald-Gruber C, Schmelzer B, Zavec D, Gasser B, Altmann F, Mattanovich D Tags: Biotechnol J Source Type: research