title: Effects of methanol feeding methods on chimeric alpha-amylase expression in continuous culture of Pichia pastoris.
abstract: The effects of two types of methanol feeding methods in a continuous culture of Pichia system on the cell growth and recombinant protein expression were studied using chimeric alpha-amylase as a model protein. With the feeding of methanol by a DO-stat method, the alpha-amylase concentration in the fermentation broth increased with decreasing dilution rate and reached 173 mg/l at a dilution rate of 0.013 h(-1), at which the maximum volumetric productivity of alpha-amylase was obtained. Although almost the same productivity was attained at 0.04 h(-1) with continuous methanol feeding, the alpha-amylase concentration was one third that compared with feeding by the DO-stat method, that is, 55 mg/l. Furthermore, at this dilution rate, the medium volume needed per unit time was three times that required when DO-stat was used. Therefore, continuous culture with methanol feeding by the DO-stat method may be a promising method for the production of recombinant proteins on an industrial scale by Pichia pastoris.
authors.affiliation: Graduate School of Science and Technology, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Nakano
authors.forename: Chun Yeon
authors.initials: CY
authors.lastname: Lee
authors.forename: Arei
authors.initials: A
authors.lastname: Yoshida
authors.forename: Takehiro
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Naofumi
authors.initials: N
authors.lastname: Shiomi
authors.forename: Shigeo
authors.initials: S
authors.lastname: Katoh
mesh_terms: D019149:Bioreactors; D001709:Biotechnology / Q000379:methods; D002478:Cells, Cultured; D005285:Fermentation; D015870:Gene Expression; D000432:Methanol / Q000737:chemistry*; D010843:Pichia / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D013997:Time Factors; D000516:alpha-Amylases / Q000737:chemistry*
chemical_list: D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000516:alpha-Amylases; D000432:Methanol