Effects of temperature and glycerol and methanol-feeding profiles on the production of recombinant galactose oxidase in Pichia pastoris
Artikel i vetenskaplig tidskrift, 2014

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. (c) 2014 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 30:728-735, 2014

MECHANISM

1986

EXPRESSION

Fusarium graminearum

Pichia pastoris

galactose oxidase

V183

OXIDATION

PEROXIDASE

OPTIMIZATION

methanol feeding

CONTAINING POLYSACCHARIDES

FED-BATCH

FAUL SJ

CATALYSIS

GLYCOPROTEINS

optimization

P244

DIRECTED EVOLUTION

PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE

Författare

George E Anasontzis

Chalmers, Kemi- och bioteknik, Industriell Bioteknik

Margarita Salazar Pena

Kemi- och bioteknik, Livsvetenskaper, Systembiologi

O. Spadiut

The Royal Institute of Technology (KTH)

Institut für Verfahrenstechnik

H. Brumer

The Royal Institute of Technology (KTH)

The University of British Columbia

Lisbeth Olsson

Chalmers, Kemi- och bioteknik, Industriell Bioteknik

Wallenberg Wood Science Center (WWSC)

Biotechnology Progress

8756-7938 (ISSN) 1520-6033 (eISSN)

Vol. 30 728-735

Ämneskategorier

Livsmedelsteknik

Mikrobiologi

DOI

10.1002/btpr.1878