Effects of temperature and glycerol and methanol-feeding profiles on the production of recombinant galactose oxidase in Pichia pastoris
Journal article, 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
galactose oxidase
methanol feeding
CONTAINING POLYSACCHARIDES
optimization
GLYCOPROTEINS
DIRECTED EVOLUTION
FAUL SJ
PROCEEDINGS OF THE SOCIETY FOR EXPERIMENTAL BIOLOGY AND MEDICINE
Pichia pastoris
PEROXIDASE
CATALYSIS
P244
OXIDATION
EXPRESSION
FED-BATCH
V183
OPTIMIZATION
MECHANISM
1986
Fusarium graminearum
Author
George E Anasontzis
Chalmers, Chemical and Biological Engineering, Industrial biotechnology
Margarita Salazar Pena
Chalmers, Chemical and Biological Engineering, Life Sciences
O. Spadiut
Royal Institute of Technology (KTH)
Institut für Verfahrenstechnik
H. Brumer
University of British Columbia (UBC)
Royal Institute of Technology (KTH)
Lisbeth Olsson
Wallenberg Wood Science Center (WWSC)
Chalmers, Chemical and Biological Engineering, Industrial biotechnology
Biotechnology Progress
8756-7938 (ISSN) 1520-6033 (eISSN)
Vol. 30 3 728-735Subject Categories
Food Engineering
Microbiology
DOI
10.1002/btpr.1878