Expression of Mycobacterium tuberculosis cytochrome CYP51 gene in Escherichia coli

Shavkynov A.S.1 , Lazarev V.N.1, Zgoda V.G.2, Govorun V.M.1, Lewi P.3, Janssen P.3, Archakov A.I.2

1. Institute of Physico-Chemical Medicine
2. Orekhovich Institute of Biomedical Chemistry RAMS
3. Center for Molecular Design
Section: Experimental/Clinical Study
PubMed Id: 14565078
Year: 2003  Volume: 49  Issue: 2  Pages: 145-152
Mycobacterium tuberculosis sterol 14a-demethylase CYP51 is currently considered a promising target for a new generation of antitubercular drugs. Similarly to eucaryotic sterol 14a-demethylases, candidate selective inhibitors of M. tuberculosis CYP51 belong to azoles, the N-substituted derivatives of imidazole and triazole. We have constructed a highly effective system for heterologous expression of MT CYP51 in E. coli. The recombinant protein was purified by metalloaffinity chromatography for primary in vitro screening of novel antitubercular azole compounds for their high affinity to the molecular target. Yield of the purified recombinant MT CYP51 was about 1,5 micromoles of the purified native protein per 1 l of E.coli cell culture. The recombinant protein interacted with ketoconazole with a Kd of 7,7 mМ. MT CYP51 produced by our system for heterologous expression in E. coli may be used for initial testing of novel antimycobacterial azole drugs.
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Shavkynov, A. S., Lazarev, V. N., Zgoda, V. G., Govorun, V. M., Lewi, P., Janssen, P., Archakov, A. I. (2003). Expression of Mycobacterium tuberculosis cytochrome CYP51 gene in Escherichia coli. Biomeditsinskaya Khimiya, 49(2), 145-152.
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