Abstract
Scale-up from shake flasks to bioreactors allows for the more reproducible, high-yielding production of recombinant proteins in yeast. The ability to control growth conditions through real-time monitoring facilitates further optimization of the process. The setup of a 3-L stirred-tank bioreactor for such an application is described.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Macauley-Patrick S, Fazenda ML, McNeil B, Harvey LM (2005) Heterologous protein production using the Pichia pastoris expression system. Yeast 22:249–270
Chen Y, Cino J, Hart G, Freedman G, White C, Komives E (1996) High protein expression in fermentation of recombinant Pichia pastoris by a fed-batch process. Process Biochem 32:107–111
Hjersted JL, Henson MA (2006) Optimization of fed-batch Saccharomyces cerevisiae fermentation using dynamic flux balance models. Biotechnol Prog 5:1239–1248
Holmes WJ, Darby RAJ, Wilks MDB, Smith R, Bill RM (2009) Developing a scalable model of recombinant protein yield from Pichia pastoris: the influence of culture conditions, biomass and induction regime. Microb Cell Fact 8:35
Zhang W, Hywood Potter K, Plantz B, Schlegel V, Smith L, Meagher M (2003) Pichia pastoris fermentation with mixed-feeds of glycerol and methanol: growth kinetics and production improvement. J Ind Microbiol Biotechnol 30:210–215
Calik P, Bayraktar E, Inankur B, Soyaslan E, Sahin M, Taspinar H, Acik E, Yilmaz R, Özdamar T (2010) Influence of pH on recombinant human growth hormone production by Pichia pastoris. Chem Technol Biotechnol 85:1628–1635
Fraser NJ (2006) Expression and functional purification of a glycosylation deficient version of the human adenosine 2a receptor for structural studies. Protein Expr Purif 49:129–137
Jamshad M, Rajesh S, Stamataki Z, McKeating JA, Dafforn TR, Overduin M, Bill RM (2008) Structural characterization of recombinant human CD81 produced in Pichia pastoris. Protein Expr Purif 57:206–216
Dragosits M, Stadlmann J, Albiol J, Baumann K, Maurer M, Gasser B, Sauer M, Altmann F, Ferrer P, Mattanovich D (2009) The effect of temperature on the proteome of recombinant Pichia pastoris. J Proteome Res 8:1380–1392
Shuler ML, Karagi F (2008) Bioprocess engineering basic concepts, 2 edn. Prentice Hall International Edition
Invitrogen: EasySelect Pichia expression kit
Invitrogen: Pichia fermentation process guidelines
O’Brien B, Britt K, Audino D (2006) Applikon bioreactor operation SOP
Routledge SJ, Hewitt CJ, Bora N, Bill RM (2011) Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield. Microb Cell Fact 10:17
Nyblom M, Öberg F, Petersson-Lindkvist K, Findlay H, Wikström J, Karlsson A, Hansson O, Booth PJ, Bill RM, Neutze R, Hedfalk K (2007) Exceptional overproduction of a functional human membrane protein. Protein Expr Purif 56:110–120
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+business Media, LLC
About this protocol
Cite this protocol
Routledge, S.J., Clare, M. (2012). Setting Up a Bioreactor for Recombinant Protein Production in Yeast. In: Bill, R. (eds) Recombinant Protein Production in Yeast. Methods in Molecular Biology, vol 866. Humana Press. https://doi.org/10.1007/978-1-61779-770-5_10
Download citation
DOI: https://doi.org/10.1007/978-1-61779-770-5_10
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-61779-769-9
Online ISBN: 978-1-61779-770-5
eBook Packages: Springer Protocols