title: Improvement of porcine interferon-α production by recombinant Pichia pastoris via induction at low methanol concentration and low temperature.
abstract: Improved porcine interferon-α (pIFN-α) production by recombinant Pichia pastoris was achieved by culture conditions optimization in a 5-l bioreactor. The results indicated that the pIFN-α concentration, specific methanol consumption rate, specific activities of alcohol oxidase, formaldehyde dehydrogenase, and formate dehydrogenase could be significantly enhanced by decreasing induction temperature. The highest pIFN-α concentration (1.35 g l(-1)) was obtained by simultaneously controlling methanol concentration at 5 g l(-1) and induction temperature at 20 °C, which was about 1.6-fold higher than the maximum obtained with previous optimal methanol concentration level (about 10 g l(-1)) when inducing at 30 °C. The potential mechanisms behind low temperature and low methanol concentration effect on pIFN-α production may be ascribed to higher cell metabolic activity, more carbon flux towards pIFN-α production, and less intracellular/extracellular protease release.
authors.affiliation: Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
authors.forename: Hu
authors.initials: H
authors.lastname: Jin
authors.forename: Guoqiang
authors.initials: G
authors.lastname: Liu
authors.forename: Keke
authors.initials: K
authors.lastname: Dai
authors.forename: Huihui
authors.initials: H
authors.lastname: Wang
authors.forename: Zhen
authors.initials: Z
authors.lastname: Li
authors.forename: Zhongping
authors.initials: Z
authors.lastname: Shi
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D000818:Animals; D019149:Bioreactors; D003470:Culture Media; D005285:Fermentation; D005560:Formate Dehydrogenases / Q000378:metabolism; D007218:Industrial Microbiology / Q000379:methods*; D016898:Interferon-alpha / Q000096:biosynthesis*; D000432:Methanol / Q000378:metabolism* / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013552:Swine; D013696:Temperature; D015533:Transcriptional Activation
chemical_list: D003470:Culture Media; D016898:Interferon-alpha; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D005560:Formate Dehydrogenases; D000445:Aldehyde Oxidoreductases; C087006:formaldehyde dehydrogenase, glutathione-independent; D000432:Methanol