Scale-up of a high cell density continuous culture with Pichia pastoris X-33 for the constitutive expression of rh-chitinase
| Autor: | Jason C. Goodrick, Nick C. Wan, Bernhard M. Schilling |
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| Rok vydání: | 2001 |
| Předmět: |
Chromatography
Chemical oxygen demand Chitinases Continuous stirred-tank reactor Biology biology.organism_classification Yeast Pichia Recombinant Proteins Dilution Pichia pastoris Oxygen Industrial Microbiology Glucose Biochemistry Chitinase Fermentation Bioreactor biology.protein Humans Air sparging Cell Division Biotechnology |
| Zdroj: | Biotechnology progress. 17(4) |
| ISSN: | 8756-7938 |
| Popis: | The feasibility of large-scale production of recombinant human chitinase using a constitutive Pichia pastoris expression system was demonstrated in a 21-L continuous stirred tank reactor. A steady-state recombinant protein concentration of 250 mg/L in the supernatant was sustained for 1 month at a dilution rate of 0.042 h -1 (equivalent to one volume exchange per day), enabling a volumetric productivity of 144 mg/L d (240 U/L d). The steady-state dry cell weight concentration in this high cell density culture reached 110 g/L. Considering safety and economical aspects, all large-scale cultivations were conducted without molecular oxygen supplementation. Conventional air sparging was used instead. The oxygen demand of the process was determined by off-gas analysis (OUR = 4.8 g O 2 L -1 h -1 with k L a = 846 h -1 ) and evaluated with regard to further reactor scale-up. |
| Databáze: | OpenAIRE |
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