Fermentation and recovery of the EcoRl restriction enzyme with a genetically modified Escherichia coli strain
The fermentation and recovery of the EcoRl restriction endonuclease with a genetically modified Escherichia coli strain is investigated. Vast amounts of product could be obtained after cultivation in a 20-L computer-coupled pilot fermentor and purification of the recovered wet cells. It was found that in the end the product is at least inhibitory and probably lethal to the cells (the lethality has been proven with genetic experiments) so that optimum yield requires an optimized choice for the time instant of induction. Growth after induction and product formation require substantial amounts of oxyge, which must be supplied if a high population level is to be achieved. pH control may alleviate the burden of high oxygen supply. Quantitative assessment after the different purification stages indicate that approximately 15% active enzyme can be obtained from the total amount produced.
- Research Organization:
- Seminar for Applied Mathematics and Biometrics, University of Ghent, Coupure links, Ghent
- OSTI ID:
- 6140455
- Journal Information:
- Biotechnol. Bioeng.; (United States), Vol. 27:9, Issue 9
- Country of Publication:
- United States
- Language:
- English
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NUCLEASES
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BIOREACTORS
CHEMICAL REACTION YIELD
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ESCHERICHIA COLI
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PH VALUE
BACTERIA
BIOCONVERSION
ENRICHMENT
ENZYMES
ESTERASES
HYDROLASES
MICROORGANISMS
PHOSPHODIESTERASES
YIELDS
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