High-cell-density fermentation of recombinant Saccharomyces cerevisiae using glycerol.
IR@CFTRI: CSIR-Central Food Technological Research Institute, Mysore
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| Relation |
http://ir.cftri.com/2095/
BP-02-02 |
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| Title |
High-cell-density fermentation of recombinant Saccharomyces cerevisiae using glycerol.
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| Creator |
Eugene Raj, A.
Sathish Kumar, H. S. Umesh Kumar, S. Misra, M. C. Ghildyal, N. P. Karanth, N. G. |
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| Subject |
04 Fermentation Technology
19 Yeast |
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| Description |
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.
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| Date |
2002
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| Type |
Article
PeerReviewed |
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| Format |
application/pdf
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| Language |
en
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| Rights |
—
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| Identifier |
http://ir.cftri.com/2095/1/Biotechnol._Prog._2002%2C_18%2C_1130-1132.pdf
Eugene Raj, A. and Sathish Kumar, H. S. and Umesh Kumar, S. and Misra, M. C. and Ghildyal, N. P. and Karanth, N. G. (2002) High-cell-density fermentation of recombinant Saccharomyces cerevisiae using glycerol. Biotechnology progress, 18 (5). pp. 1130-2. ISSN 8756-7938 |
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