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Human xanthine oxidase recombinant in E. coli: a whole cell catalyst for preparative drug metabolites synthesis

Ferreira Antunes, Marcia and Eggimann, Fabian and Kittelmann, Matthias and Luetz, Stephan and Hanlon, Steven Paul and Wirz, Beat and Bachler, Thorsten and Winkler, Margit (2016) Human xanthine oxidase recombinant in E. coli: a whole cell catalyst for preparative drug metabolites synthesis. Journal of Biotechnology. ISSN 01681656

Abstract

Human xanthine oxidoreductase (XOR), which is responsible for the final steps of the purine metabolism pathway and involved in oxidative drug metabolism, was successfully expressed in Escherichia coli BL21(DE3) Gold. Recombinant human (rh) XOR yielded higher productivity with the gene sequence optimized for expression in E.coli than with the native gene sequence. Induction of XOR expression with lactose or IPTG resulted in complete loss of activity whereas shake flasks cultures using media rather poor in nutrients resulted in functional XOR expression in the stationary phase. LB medium was used for a 25 L fermentation in fed-batch mode, which led to a 5 fold increase of the enzyme yield when compared to production in shake flasks. Quinazoline was used as a substrate on the semi-preparative scale using an optimized whole cell biotransformation protocol, yielding 73 mg of the isolated product, 4-quinazolinone, from 100 mg of starting material.

Item Type: Article
Keywords: Biocatalysis, xanthine oxidase, heterologous expression, E. coli, drug metabolite synthesis
Date Deposited: 26 Apr 2016 23:45
Last Modified: 26 Apr 2016 23:45
URI: https://oak.novartis.com/id/eprint/27413

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