Improved production and stability ofE. coli recombinants expressing transketolase for large scale biotransformation |
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Authors: | Carol French John M. Ward |
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Affiliation: | (1) Department of Biochemistry and Molecular Biology, University College London, Gower Street, WC1E 6BT London, UK |
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Abstract: | Summary TheE.coli tkt gene has been subcloned into high copy number vectors. In fed batch fermentations up to 4gL–1 of soluble intracellular transketolase was produced representing 43% of the total cell protein. Increased plasmid stability during fed-batch fermentations was obtained by using kanamycin resistant pBGS vectors rather than the ampicillin resistant pUC vectors. Plasmid stability was maintained throughout growth in a complex medium without any selective pressure by incorporating thecer region fromColE1 into the expression construct. |
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