US EPA

1043724 

Journal Article 

Production of recombinant protein in Pichia pastoris by fermentation 

Tolner, B; Smith, L; Begent, RH; Chester, KA 

2006 

1 

Nature Protocols
ISSN: 1754-2189
EISSN: 1750-2799 

Nature Publishing Group, The Macmillan Building 4 Crinan Street London N1 9XW UK, [mailto:feedback@nature.com], [URL:http://www.nature.com/] 

1 

2 

1006-1021 

English 

17406338 

10.1038/nprot.2006.126 

WOS:000251002500068 

This protocol is applicable to recombinant protein expression by small-scale fermentation using the Pichia pastoris expression system. P. pastoris has the capacity to produce large quantities of protein with eukaryotic processing. Expression is controlled by a methanol-inducible promoter, which allows a biomass-generation phase before protein production is initiated. The target protein is secreted directly into a protein-free mineral salt medium, and is relatively easy to purify. The protocol is readily interfaced with expanded bed adsorption for immediate capture and purification of recombinant protein. The setting up of the bioreactor plus the fermentation itself takes 1 wk. Making the master and user seed lots takes approximately 2 wk for each individual clone. 

Promoters; Adsorption; protein purification; Seeds; Fermentation; Pichia pastoris; Bioreactors; Minerals; Salts