title: Identification of a gene involved in the synthesis of a dipeptidyl peptidase IV inhibitor in Aspergillus oryzae.
abstract: WYK-1 is a dipeptidyl peptidase IV inhibitor produced by Aspergillus oryzae strain AO-1. Because WYK-1 is an isoquinoline derivative consisting of three l-amino acids, we hypothesized that a nonribosomal peptide synthetase was involved in its biosynthesis. We identified 28 nonribosomal peptide synthetase genes in the sequenced genome of A. oryzae RIB40. These genes were also identified in AO-1. Among them, AO090001000009 (wykN) was specifically expressed under WYK-1-producing conditions in AO-1. Therefore, we constructed wykN gene disruptants of AO-1 after nonhomologous recombination was suppressed by RNA interference to promote homologous recombination. Our results demonstrated that the disruptants did not produce WYK-1. Furthermore, the expression patterns of 10 genes downstream of wykN were similar to the expression pattern of wykN under several conditions. Additionally, homology searches revealed that some of these genes were predicted to be involved in WYK-1 biosynthesis. Therefore, we propose that wykN and the 10 genes identified in this study constitute the WYK-1 biosynthetic gene cluster.
authors: Healthcare Research Institute, Wakunaga Pharmaceutical Co. Ltd., Akitakata, Japan. imamura_kz@wakunaga.co.jp
authors: Kazuhiko
authors: K
authors: Imamura
authors: Yoshihito
authors: Y
authors: Tsuyama
authors: Terukage
authors: T
authors: Hirata
authors: Sumihiro
authors: S
authors: Shiraishi
authors: Kazutoshi
authors: K
authors: Sakamoto
authors: Osamu
authors: O
authors: Yamada
authors: Osamu
authors: O
authors: Akita
authors: Hitoshi
authors: H
authors: Shimoi
mesh_terms: D001236:Aspergillus oryzae / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism*; D053898:Biosynthetic Pathways / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D018819:Dipeptidyl Peptidase 4 / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism*; D017353:Gene Deletion; D007546:Isoquinolines / Q000378:metabolism*; D008969:Molecular Sequence Data; D005810:Multigene Family; D010453:Peptide Synthases / Q000235:genetics / Q000378:metabolism*; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal; D004791:Enzyme Inhibitors; D007546:Isoquinolines; D018819:Dipeptidyl Peptidase 4; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase; C039109:isoquinoline