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Production of Recombinant Human Aldehyde Oxidase in Escherichia coli and Optimization of Its Application for the Preparative Synthesis of Oxidized Drug Metabolites
Author(s) -
Rodrigues Diogo,
Kittelmann Matthias,
Eggimann Fabian,
Bachler Thorsten,
Abad Sandra,
Camattari Andrea,
Glieder Anton,
Winkler Margit,
Lütz Stephan
Publication year - 2014
Publication title -
chemcatchem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.497
H-Index - 106
eISSN - 1867-3899
pISSN - 1867-3880
DOI - 10.1002/cctc.201301094
Subject(s) - biotransformation , bioreactor , chemistry , escherichia coli , yield (engineering) , biochemistry , bioconversion , chromatography , aldehyde oxidase , commodity chemicals , metabolic engineering , fermentation , enzyme , organic chemistry , catalysis , xanthine oxidase , materials science , metallurgy , gene
Recombinant human aldehyde oxidase (AO) was expressed in Escherichia coli . Different cell disruption methods and conditions of cell culture in shake flasks and bioreactors and of biotransformation on an analytical scale were tested to optimize the synthesis of oxidized AO drug metabolites. The volumetric productivity was increased 24‐fold by optimizing the cell culture conditions. The highest yield was achieved in a 25 L stirred tank bioreactor under non‐oxygen‐limited conditions and high lactose feed rate. Suspensions of highly concentrated and well‐aerated whole cells at neutral pH and relatively low temperatures led to the best conversion. The solvent for the substrate and the buffering agent for the biotransformation had an important effect. In a biotransformation with AO, 210 mg of famciclovir was converted to diacetyl penciclovir a yield of 82 %. The optimized protocol represents a viable method for the preparative synthesis of oxidized AO metabolites of drugs.