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Title
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Recombinant Escherichia coli cells immobilized in Ca-alginate beads for metabolite production
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Author
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Abstract
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| Milligram amounts of metabolites of drug candidates arc required to identify toxic products. Human drug metabolites are currently produced selectively in a time- and cost-efficient manner in bioreactor systems containing recombinant Escherichia coli co-expressing a human cytochrome P450 isoenzyme/NADPH cytochrome P450 reductase (hCYP/HR) complex. For further optimization, immobilization of the catalytic system in Ca-alginate microbeads was considered. This new concept was designed for CYP3A4 with testosterone as substrate. Immobilized E. coli cells had a high maximal and homogeneously distributed biomass. Viability was stable over at least I week of culture and even longer during storage. Gene expression was ideally initiated 6 h after immobilization. Although immobilized E. coli cells expressed a highly functional enzyme system after 2 days, they did not metabolize testosterone, probably due to cell permeability problems resulting from immobilization. Therefore, immobilized cell membranes displaying testosterone bioconversion activity, even after long-term storage, will be used in bioreactors with high organic solvent content. |
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Language
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English
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Source (journal)
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Biocatalysis and biotransformation. - Basel
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Publication
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Basel
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2009
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ISSN
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1024-2422
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DOI
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10.3109/10242420903286141
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Volume/pages
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27
:5/6
(2009)
, p. 348-359
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ISI
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000272087700009
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Full text (Publisher's DOI)
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