PMID- 12790641
OWN - NLM
STAT- MEDLINE
DCOM- 20040524
LR  - 20210308
IS  - 8756-7938 (Print)
IS  - 1520-6033 (Linking)
VI  - 19
IP  - 3
DP  - 2003 May-Jun
TI  - Improved production of recombinant ovine interferon-tau by mut(+) strain of 
      Pichia pastoris using an optimized methanol feed profile.
PG  - 794-802
AB  - Recombinant ovine interferon-tau (r-oIFN-tau) production by Pichia pastoris was 
      studied using methanol as the sole carbon source during induction. The cells were 
      grown on glycerol up to a certain cell density before induction of the AOX1 
      promoter by methanol for expression of the recombinant protein. Cell growth on 
      methanol has been modeled using a substrate-feed equation, which served as the 
      basis for an effective computer control of the process. The r-oIFN-tau 
      concentration in the culture began to decline despite continued cell growth after 
      50 (+/- 6) h of induction, which was associated with an increase in proteolytic 
      activity of the fermentation broth. A specific growth rate of 0.025 h(-1) was 
      found to be optimal for r-oIFN-tau production. No significant improvement in 
      r-oIFN-tau production was observed when the specific growth rate was stepped up 
      before the critical point when r-oIFN-tau concentration started decreasing during 
      fermentation. However, best results were obtained when the specific growth rate 
      was stepped down from 0.025 to 0.02 h(-1) at 38 h of induction, whereby the 
      active production period was prolonged until 70 h of induction and the broth 
      protease activity was correspondingly reduced. The corresponding maximum protein 
      yield was 391.7 mg x L(-1) after 70 h of fermentation. The proteolytic activity 
      could be reduced by performing fermentations at specific growth rates of 0.025 
      h(-1) or below. The recombinant protein production can be performed at an optimal 
      yield by directly controlling the methanol feed rate by a computer-controlled 
      model. The production profile of r-oIFN-tau was found to be significantly 
      different from other secreted and intracellular recombinant protein processes, 
      which is an indication that recombinant protein production in Pichia pastoris 
      needs to be optimized as individual processes following established principles.
FAU - Sinha, Jayanta
AU  - Sinha J
AD  - Biological Process Development Facility, Department of Chemical Engineering, 
      University of Nebraska-Lincoln, Lincoln, Nebraska 68583, USA.
FAU - Plantz, Bradley A
AU  - Plantz BA
FAU - Zhang, Wenhui
AU  - Zhang W
FAU - Gouthro, Mark
AU  - Gouthro M
FAU - Schlegel, Vicki
AU  - Schlegel V
FAU - Liu, Chih-Ping
AU  - Liu CP
FAU - Meagher, Michael M
AU  - Meagher MM
LA  - eng
PT  - Comparative Study
PT  - Evaluation Study
PT  - Journal Article
PT  - Validation Study
PL  - United States
TA  - Biotechnol Prog
JT  - Biotechnology progress
JID - 8506292
RN  - 0 (Interferon Type I)
RN  - 0 (Pregnancy Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (interferon tau)
RN  - Y4S76JWI15 (Methanol)
SB  - IM
MH  - Animals
MH  - Bioreactors/*microbiology
MH  - Cell Culture Techniques/*methods
MH  - Cell Division/physiology
MH  - Feedback
MH  - Interferon Type I/*biosynthesis/genetics/isolation & purification
MH  - Methanol/*metabolism
MH  - *Models, Biological
MH  - Pichia/cytology/genetics/*growth & development/*metabolism
MH  - Pregnancy Proteins/*biosynthesis/genetics/isolation & purification
MH  - Protein Engineering/*methods
MH  - Quality Control
MH  - Recombinant Proteins/biosynthesis
MH  - Sheep
EDAT- 2003/06/07 05:00
MHDA- 2004/05/25 05:00
CRDT- 2003/06/07 05:00
PHST- 2003/06/07 05:00 [pubmed]
PHST- 2004/05/25 05:00 [medline]
PHST- 2003/06/07 05:00 [entrez]
AID - 10.1021/bp025744q [doi]
PST - ppublish
SO  - Biotechnol Prog. 2003 May-Jun;19(3):794-802. doi: 10.1021/bp025744q.