High-Cell-Density Fermentation of Recombinant Saccharomyces cerevisiae Using Glycerol

To obtain a high cell density of recombinant Saccharomyces cerevisiae (INVSc 1 strain bearing a 2 μm plasmid, pYES2 containing a GAL1 promoter for expression of the β‐galactosidase gene), the yeast was grown with glycerol as the substrate by fed‐batch fermentation. The feeding strategy was based on...

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Veröffentlicht in:Biotechnology progress 2002, Vol.18 (5), p.1130-1132
Hauptverfasser: Raj, A. Eugene, Kumar, H. S. Sathish, Kumar, S. Umesh, Misra, M. C., Ghildyal, N. P., Karanth, N. G.
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container_end_page 1132
container_issue 5
container_start_page 1130
container_title Biotechnology progress
container_volume 18
creator Raj, A. Eugene
Kumar, H. S. Sathish
Kumar, S. Umesh
Misra, M. C.
Ghildyal, N. P.
Karanth, N. G.
description To obtain a high cell density of recombinant Saccharomyces cerevisiae (INVSc 1 strain bearing a 2 μm plasmid, pYES2 containing a GAL1 promoter for expression of the β‐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.
doi_str_mv 10.1021/bp0201105
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subjects Biological and medical sciences
Bioreactors
Cell Count
Feedback
Fermentation
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Fungal
Glucose - metabolism
Glycerol - metabolism
Hydrogen-Ion Concentration
Phosphates - metabolism
Recombination, Genetic
Saccharomyces cerevisiae - classification
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Sensitivity and Specificity
Species Specificity
title High-Cell-Density Fermentation of Recombinant Saccharomyces cerevisiae Using Glycerol
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