Journal
BIOPROCESS AND BIOSYSTEMS ENGINEERING
Volume 33, Issue 4, Pages 473-483Publisher
SPRINGER
DOI: 10.1007/s00449-009-0356-3
Keywords
Fed-batch culture; Multi-variables clustering; Oxygen uptake rate (OUR); Pichia pastoris; Porcine interferon-alpha expression
Funding
- Shanghai Science & Technology Committee [073919108]
- National High-tech RD program [2006AA020101]
- National Natural Science foundation [60604028]
- Major State Basic Research Development Program, of China [2007CB714303]
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Efficient porcine interferon-alpha (pIFN-alpha) expression in high density recombinant Pichia pastoris cultivation was achieved in a 5 l bench-scaled bioreactor. The results indicated that a high and stable oxygen uptake rate (OUR) during induction phase was closely related with pIFN-alpha production efficiency. The multi-variables clustering and analysis results showed that the achievement of a high and stable OUR relied on a higher glycerol consumption rate during fed-batch culture phase and a moderate methanol level (around 10 g/l) during induction phase. In the high and stable OUR environments (200-300 mmol/l/h), the highest pIFN-alpha antiviral activity could reach a level of 6.7 x 10(6) IU/ml, which was more than 10-300-folds higher than those obtained at lower OUR (80-200 mmol/l/h) using the same bioreactor and those obtained in shaking flasks. Clustering and analysis of the specific growth and glycerol consumption rates data during culture phase could detect the ill fermentation state at early stage, potentially provided a simple and effective fault alarming/diagnosis method for the achievement of stable pIFN-alpha production.
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