PMID- 15580575
OWN - NLM
STAT- MEDLINE
DCOM- 20050428
LR  - 20220317
IS  - 0006-3592 (Print)
IS  - 0006-3592 (Linking)
VI  - 89
IP  - 1
DP  - 2005 Jan 5
TI  - Causes of proteolytic degradation of secreted recombinant proteins produced in 
      methylotrophic yeast Pichia pastoris: case study with recombinant ovine 
      interferon-tau.
PG  - 102-12
AB  - It was observed that during fermentative production of recombinant ovine 
      interferon-tau (r-oIFN-tau) 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.
CI  - (c) 2004 Wiley Periodicals, Inc.
FAU - Sinha, Jayanta
AU  - Sinha J
AD  - Biological Process Development Facility, Department of Chemical Engineering, 
      University of Nebraska-Lincoln, NE, USA. jsinha@unlnotes.unl.edu
FAU - Plantz, Bradley A
AU  - Plantz BA
FAU - Inan, Mehmet
AU  - Inan M
FAU - Meagher, Michael M
AU  - Meagher MM
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
RN  - 0 (Interferon Type I)
RN  - 0 (Pregnancy Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (interferon tau)
RN  - EC 3.4.- (Peptide Hydrolases)
RN  - PDC6A3C0OX (Glycerol)
RN  - Y4S76JWI15 (Methanol)
SB  - IM
MH  - Animals
MH  - Bioreactors/*microbiology
MH  - Blotting, Western
MH  - Cell Fractionation
MH  - Cytoplasm/enzymology
MH  - Glycerol/metabolism
MH  - Interferon Type I/*biosynthesis/genetics/isolation & purification
MH  - Kinetics
MH  - Methanol/*metabolism
MH  - Peptide Hydrolases/metabolism
MH  - Pichia/cytology/genetics/*growth & development/*metabolism
MH  - Pregnancy Proteins/*biosynthesis/genetics/isolation & purification
MH  - Recombinant Proteins/metabolism
MH  - Sheep
MH  - Temperature
MH  - Time Factors
MH  - Vacuoles/enzymology
EDAT- 2004/12/08 09:00
MHDA- 2005/04/29 09:00
CRDT- 2004/12/08 09:00
PHST- 2004/12/08 09:00 [pubmed]
PHST- 2005/04/29 09:00 [medline]
PHST- 2004/12/08 09:00 [entrez]
AID - 10.1002/bit.20318 [doi]
PST - ppublish
SO  - Biotechnol Bioeng. 2005 Jan 5;89(1):102-12. doi: 10.1002/bit.20318.