High-cell-density fermentation of recombinant Saccharomyces cerevisiae using glycerol
| Autor: | N. P. Ghildyal, Manoj Misra, A. Eugene Raj, H. S. Sathish Kumar, N. G. Karanth, S. Umesh Kumar |
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| Rok vydání: | 2002 |
| Předmět: |
Glycerol
Ammonium phosphate Saccharomyces cerevisiae Biomass Cell Count Sensitivity and Specificity Feedback Phosphates chemistry.chemical_compound Bioreactors Species Specificity Gene Expression Regulation Fungal Food science Recombination Genetic biology Cell growth Substrate (chemistry) Hydrogen-Ion Concentration biology.organism_classification Yeast Glucose chemistry Biochemistry Fermentation Biotechnology |
| Zdroj: | Biotechnology progress. 18(5) |
| ISSN: | 8756-7938 |
| Popis: | To obtain a high cell density of recombinant Saccharomyces cerevisiae (INVSc 1 strain bearing a 2 microm plasmid, pYES2 containing a GAL1 promoter for expression of the beta-galactosidase gene), the yeast was grown with glycerol as the substrate by fed-batch fermentation. The feeding strategy was based on an on-line response of the medium pH to the consumption of glycerol. The approach was to feed excess carbon into the medium to create a benign environment for rapid biomass buildup. During cell growth in the presence of glycerol, the release of protons in the medium caused a decrease in pH and the consumption rate of ammonium phosphate served as an on-line indicator for the metabolic rate of the organism. The extent of glycerol feeding in a fed-batch mode with pH control at 5.0 +/- 0.1 was ascertained from the automatic addition of ammonium phosphate to the medium. The glycerol feeding to ammonium phosphate addition ratio was found to be 2.5-3.0. On the basis of the experiments, a maximum dry cell biomass of 140 g per liter and a productivity of 5.5 g DCW/L/h were achieved. The high cell density of S. cerevisiae obtained with good plasmid stability suggested a simple and efficient fermentation protocol for recombinant protein production. |
| Databáze: | OpenAIRE |
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