Characterization and functional analysis of a novel multicopper oxidase and associated polyketide biosynthesis gene cluster of aspergillus fumigatus

Metin, Banu
In this study, novel polyketide biosynthesis gene cluster of Aspergillus fumigatus was characterized and functionally analyzed. Analysis of the newly sequenced A. fumigatus genome for laccases, which are involved in melanin biosynthesis and detoxification in fungi, resulted in several putative laccase and multicopper oxidase gene sequences, one of which, Afu4g14490 (tpnJ), was selected for further characterization. The predicted amino acid sequence TpnJp showed 63% identity with the dihydrogeodin oxidase of Aspergillus terreus, which is involved in the biosynthesis of the antifungal geodin. When the genome region of tpnJ was investigated, the presence of a polyketide biosynthesis gene cluster containing 13 genes, hypothesized to be responsible for the production of trypacidin and monomethylsulochrin, was realized. By a comparative genomics approach, a putative geodin biosynthesis gene cluster containing 13 genes, including dihydrogeodin oxidase, in A. terreus and a putative trypacidin biosynthesis gene cluster containing 13 genes in N. fischeri were established. Targeted deletions of the polyketide synthase (tpnC) and multicopper oxidase (tpnJ) genes confirmed the hypothesis that TpnCp, a three-domain minimal polyketide synthase, is involved in trypacidin and monomethylsulochrin biosynthesis in A. fumigatus. TpnCp is the first fungal minimal polyketide synthase whose functional role was experimentally identified. Moreover, the fact that LC-MS analysis of DtpnJ strain showed the absence of trypacidin and the presence of a higher amount of monomethylsulochrin in DtpnJ strain, confirmed the hypothesis that TpnJp is involved in the oxidation of monomethylsulochrin into trypacidin. This novel multicopper oxidase having high substrate specificity is given the name monomethylsulochrin oxidase.


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Citation Formats
B. Metin, “Characterization and functional analysis of a novel multicopper oxidase and associated polyketide biosynthesis gene cluster of aspergillus fumigatus,” Ph.D. - Doctoral Program, Middle East Technical University, 2007.