High-cell-density fermentation studies of a recombinantE. coli that expresses atrial natriuretic factor |
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Authors: | Dr F James Bailey Janet Blankenship Jon H Condra Robert Z Maigetter Ronald W Ellis |
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Institution: | (1) Dept. of Virus and Cell Biology, Merck Sharp & Dohme Research Laboratories, 19486 West Point, PA, U.S.A. |
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Abstract: | Summary Studies are presented on the fermentation of recombinantEscherichia coli that express rat atrial natriuretic factor (ANF) as a fusion protein. Our objective was to achieve high cell density while maintaining ANF expression at the same level as observed in shake flasks. Improved fermentation conditions included: maintaining glucose concentrations at 1 g/l, using an enriched medium, adding concentrates of medium throughout the fermentation, and blending oxygen for adequate aeration. Cell densities of 12 g/l (dry weight) were achieved, which represented a 10-fold increase over non-improved conditions, while maintaining ANF levels at 7 mg/g of dry cell mass. When galactose was used as an initial carbon source or as a feed supplement, there was a 2-3-fold increase in the expression of ANF from these high-cell-density fermentations. The recombinant ANF was biologically active. |
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Keywords: | Fermentation High cell density Escherichia coli RecombinantE coli Atrial natriuretic factor (ANF) Fusion protein expression |
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