Expression system for recombinant proteins

C - Chemistry – Metallurgy – 12 – N

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C12N 15/81 (2006.01) C12N 9/16 (2006.01) C12N 9/18 (2006.01) C12N 9/24 (2006.01) C12P 21/02 (2006.01)

Patent

CA 2429210

A continuous fermentation process has been developed in Pichia pastoris (P. pastoris) is order to produce large quantities of recombinant human proteins. High expression levels have been demonstrated in continuous production of the enzyme by P. pastoris with a constitutive promoter in a 1.5-liter working volume fermenter using either glucose or glycerol as the carbon source. The fermentation could be extended for long periods of time with an excellent steady-state protein concentration and cell densities achieved. No proteolytic degradation of the enzyme was seen in the continuous fermentation mode.

L'invention porte sur un procédé de fermentation continu qui a été mis au point dans la levure Pichia pastoris (P.pastoris) afin de produire des quantités élevées de protéines humaines recombinées. Des niveaux d'expression élevés sont apparus durant la production continue de l'enzyme par la levure P.pastoris à l'aide d'un agent promoteur constitutif dans un fermenteur de volume de travail de 1,5 litres en utilisant du glucose ou du glycérol comme source de carbone. La fermentation peut être prolongée sur une longue période avec la possibilité d'obtenir un excellent niveau de concentration de protéines à état stable et de densités cellulaires. Aucune dégradation protéolytique de l'enzyme n'a été constatée au cours du mode continu de fermentation.

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