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Molecular characterization and isolation of cytochrome P450 genes from the filamentous fungus Aspergillus oryzae

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Abstract

We explored the molecular diversity of cytochrome P450 genes in the filamentous fungus Aspergillus oryzae using bioinformatic and experimental approaches. Based on bioinformatic annotation, we found 155 putative genes of cytochromes P450 in the whole genome sequence; however, 13 of 155 appeared to be pseudogenes due to sequence deletions and/or inframe stop codon(s). There are 87 families of A. oryzae cytochromes P450 (AoCYPs), indicating considerable phylogenetic diversity. To characterize A. oryzae AoCYPs, we attempted to isolate cDNAs using RT-PCR and determined their transcriptional capabilities. To date, we have confirmed gene expression of 133 AoCYPs and cloned 121 AoCYPs as full-length cDNAs encoding a mature open reading frame. Using experimentally deduced sequences and intron–exon organization, we analyzed AoCYPs phylogenetically. We also identified intronic consensus sequences in AoCYPs genes. The experimentally validated exonic and intronic sequences will be a powerful advantage in identification and characterization of novel P450s from various ascomycetous fungi.

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Acknowledgments

We are grateful to Hiroko Tsutsumi and Yoji Hata (Res. Inst., Gekkeikan Sake Co.) for providing A. oryzae strain RIB40 and for valuable technical advices. This research was supported in part by an NIRS Research Grant (Noda Institute for Scientific Research; to H. I.).

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Authors and Affiliations

  1. Faculty of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    K. H. M. Nazmul Hussain Nazir, Hirofumi Ichinose & Hiroyuki Wariishi

  2. Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Hiroyuki Wariishi

  3. Innovation Center for Medical Redox Navigation, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan

    Hiroyuki Wariishi

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  1. K. H. M. Nazmul Hussain Nazir
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  2. Hirofumi Ichinose
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Correspondence to Hirofumi Ichinose or Hiroyuki Wariishi.

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Communicated by Axel Brakhage.

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Nazmul Hussain Nazir, K.H.M., Ichinose, H. & Wariishi, H. Molecular characterization and isolation of cytochrome P450 genes from the filamentous fungus Aspergillus oryzae . Arch Microbiol 192, 395–408 (2010). https://doi.org/10.1007/s00203-010-0562-z

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  • DOI: https://doi.org/10.1007/s00203-010-0562-z

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