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Increased resistance to 14α-demethylase inhibitors (DMIs) in Aspergillus niger by coexpression of the Penicillium italicum eburicol 14α-demethylase (cyp51) and the A. Niger cytochrome P450 reductase (cprA) genes

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Author: Brink, H.J.M. van den · Nistelrooy, H.J.G.M. van · Waard, M.A. de · Hondel, C.A.M.J.J. van den · Gorcom, R.F.M. van
Type:article
Date:1996
Institution: Prins Maurits Laboratorium TNO
Source:Journal of Biotechnology, 1-3, 49, 13-18
Identifier: 233428
doi: doi:10.1016/0168-1656(96)01403-4
Keywords: Biology · Ergosterol · NADPH cytochrome P450 reductase · Sterol 14α-demethylation inhibitors · Drug products · Fungicides · Genes · Demethylase inhibitors (DMI) · Gene encoding · Genetic engineering · Cytochrome p450 reductase · Reduced nicotinamide adenine dinucleotide phosphate ferrihemoprotein reductase · Enzyme synthesis · Fungal genetics · Nonhuman · Penicillium · Priority journal · Aspergillus niger · Cytochrome P-450 Enzyme System · Drug Resistance, Microbial · Enzyme Inhibitors · Gene Expression Regulation, Fungal · NADH, NADPH Oxidoreductases · NADPH-Ferrihemoprotein Reductase · Oxidoreductases · Penicillium · Recombinant Proteins · Transformation, Genetic · Aspergillus niger · Fungi · Penicillium · Penicillium italicum

Abstract

In this paper we describe the effects of over-expression of the Penicillium italicum gene encoding eburicol 14α-demethylase (cyp51), in Aspergillus niger strains with one or multiple copies of the gene encoding cytochrome P450 reductase (cpr A), on the eburicol 14α-demethylase activity. Eburicol 14α-demethylase activity was determined by measuring the resistance of transformants against some eburicol 14α-demethylase inhibitors (DMIs). DMIs are widely used as fungicides in crop protection and human and veterinarian health care. DMI resistance in a transformant overexpressing both CPR and CYP51 was increased 5-30-fold compared to DMI resistance in the wild type strain, depending on the test compound used. Resistance in this strain was approximately 2-5-fold increased compared to DMI resistance in a transformant that was overexpressing the cyp51 gene but had only the wild type copy of the cprA gene and approximately 3-12-fold increased compared to a strain overexpressing the cprA gene (and having only the wild type copy of the cyp51 gene). These results show the importance of CPR overexpression for increasing cytochrome P450 activities in filamentous fungi.