Effective and stable porcine interferon-alpha production by Pichia pastoris fed-batch cultivation with multi-variables clustering and analysis.
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.
Yu
Ruisong
R
Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Science, China.
Dong
Shijuan
S
Zhu
Yumin
Y
Jin
Hu
H
Gao
Mingjie
M
Duan
Zuoying
Z
Zheng
Zhiyong
Z
Shi
Zhongping
Z
Li
Zhen
Z
D000818:Animals; D000998:Antiviral Agents / Q000378:metabolism; D057005:Bioengineering; D019149:Bioreactors / Q000382:microbiology* / Q000706:statistics & numerical data; D016000:Cluster Analysis; D005285:Fermentation; D005990:Glycerol / Q000378:metabolism; D066298:In Vitro Techniques; D007370:Interferon Type I / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D000432:Methanol / Q000378:metabolism; D010101:Oxygen Consumption; D010843:Pichia / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011994:Recombinant Proteins; D013552:Swine
D000998:Antiviral Agents; D007370:Interferon Type I; D011994:Recombinant Proteins; D005990:Glycerol; D000432:Methanol