title: Effective and stable porcine interferon-alpha production by Pichia pastoris fed-batch cultivation with multi-variables clustering and analysis.
abstract: 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.
authors: Animal Husbandry and Veterinary Research Institute, Shanghai Academy of Agricultural Science, China.
authors: Ruisong
authors: R
authors: Yu
authors: Shijuan
authors: S
authors: Dong
authors: Yumin
authors: Y
authors: Zhu
authors: Hu
authors: H
authors: Jin
authors: Mingjie
authors: M
authors: Gao
authors: Zuoying
authors: Z
authors: Duan
authors: Zhiyong
authors: Z
authors: Zheng
authors: Zhongping
authors: Z
authors: Shi
authors: Zhen
authors: Z
authors: Li
mesh_terms: 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
chemical_list: D000998:Antiviral Agents; D007370:Interferon Type I; D011994:Recombinant Proteins; D005990:Glycerol; D000432:Methanol