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Causes of proteolytic degradation of secreted recombinant proteins produced in methylotrophic yeast Pichia pastoris : Case study with recombinant ovine interferon‐τ
Author(s) -
Sinha Jayanta,
Plantz Bradley A.,
Inan Mehmet,
Meagher Michael M.
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20318
Subject(s) - pichia pastoris , recombinant dna , proteases , fermentation , biochemistry , protease , biology , lysis , glycerol , protein degradation , chemistry , enzyme , gene
It was observed that during fermentative production of recombinant ovine interferon‐τ (r‐oIFN‐τ) in Pichia pastoris, a secreted recombinant protein, the protein was degraded increasingly after 48 h of induction and the rate of degradation increased towards the end of fermentation at 72 h, when the fermentation was stopped. Proteases, whose primary source was the vacuoles, was found in increasing levels in the cytoplasm and in the fermentation broth after 48 h of induction and reached maximal values when the batch was completed at 72 h. Protease levels at various cell fractions as well as in the culture supernatant were lower when glycerol was used as the carbon source instead of methanol. It can be concluded that methanol metabolism along with cell lysis towards the end of fermentation contributes to increased proteolytic activity and eventual degradation of recombinant protein. © 2004 Wiley Periodicals, Inc.