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Effects of temperature and glycerol and methanol-feeding profiles on the production of recombinant galactose oxidase in Pichia pastoris
KTH, School of Biotechnology (BIO), Glycoscience. KTH, School of Chemical Science and Engineering (CHE), Centres, Wallenberg Wood Science Center.
KTH, School of Biotechnology (BIO), Glycoscience. KTH, School of Chemical Science and Engineering (CHE), Centres, Wallenberg Wood Science Center.
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2014 (English)In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033, Vol. 30, no 3, 728-735 p.Article in journal (Refereed) Published
Abstract [en]

Optimization of protein production from methanol-induced Pichia pastoris cultures is necessary to ensure high productivity rates and high yields of recombinant proteins. We investigated the effects of temperature and different linear or exponential methanol-feeding rates on the production of recombinant Fusarium graminearum galactose oxidase (EC 1.1.3.9) in a P. pastoris Mut+ strain, under regulation of the AOX1 promoter. We found that low exponential methanol feeding led to 1.5-fold higher volumetric productivity compared to high exponential feeding rates. The duration of glycerol feeding did not affect the subsequent product yield, but longer glycerol feeding led to higher initial biomass concentration, which would reduce the oxygen demand and generate less heat during induction. A linear and a low exponential feeding profile led to productivities in the same range, but the latter was characterized by intense fluctuations in the titers of galactose oxidase and total protein. An exponential feeding profile that has been adapted to the apparent biomass concentration results in more stable cultures, but the concentration of recombinant protein is in the same range as when constant methanol feeding is employed.

Place, publisher, year, edition, pages
2014. Vol. 30, no 3, 728-735 p.
Keyword [en]
galactose oxidase, Pichia pastoris, methanol feeding, optimization, Fusarium graminearum
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:kth:diva-148259DOI: 10.1002/btpr.1878ISI: 000337628900022Scopus ID: 2-s2.0-84922969799OAI: oai:DiVA.org:kth-148259DiVA: diva2:736782
Funder
Knut and Alice Wallenberg Foundation
Note

QC 20140808

Available from: 2014-08-08 Created: 2014-08-04 Last updated: 2017-12-05Bibliographically approved

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