Title
Recombinant Escherichia coli cells immobilized in Ca-alginate beads for metabolite production Recombinant Escherichia coli cells immobilized in Ca-alginate beads for metabolite production
Author
Faculty/Department
Faculty of Sciences. Biology
Publication type
article
Publication
Basel ,
Subject
Chemistry
Biology
Engineering sciences. Technology
Source (journal)
Biocatalysis and biotransformation. - Basel
Volume/pages
27(2009) :5/6 , p. 348-359
ISSN
1024-2422
ISI
000272087700009
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
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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