title: Biosynthetic gene-based secondary metabolite screening: a new diterpene, methyl phomopsenonate, from the fungus Phomopsis amygdali.
abstract: The presence of the geranylgeranyl diphosphate synthase (GGS) gene is a common feature of gene clusters for diterpene biosynthesis. We demonstrated identification of a diterpene gene cluster using homology-based PCR of GGS genes and the subsequent genome walking in the fungus Phomopsis amygdali N2. Structure determination of a novel diterpene hydrocarbon phomopsene provided by enzymatic synthesis with the recombinant terpene synthase PaPS and screening of fungal broth extracts with reference to characteristic NMR signals of phomopsene allowed us to isolate a new diterpene, methyl phomopsenonate. The versatility of the gene-based screening of unidentified diterpenes is discussed in regard to fungal genomic data.
authors.affiliation: Department of Bioresource Engineering, Yamagata University, Tsuruoka, Yamagata 997-8555, Japan.
authors.forename: Tomonobu
authors.initials: T
authors.lastname: Toyomasu
authors.forename: Akane
authors.initials: A
authors.lastname: Kaneko
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Tokiwano
authors.forename: Yuya
authors.initials: Y
authors.lastname: Kanno
authors.forename: Yuri
authors.initials: Y
authors.lastname: Kanno
authors.forename: Rie
authors.initials: R
authors.lastname: Niida
authors.forename: Shigeyoshi
authors.initials: S
authors.lastname: Miura
authors.forename: Taiki
authors.initials: T
authors.lastname: Nishioka
authors.forename: Chiho
authors.initials: C
authors.lastname: Ikeda
authors.forename: Wataru
authors.initials: W
authors.lastname: Mitsuhashi
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
authors.forename: Tomikazu
authors.initials: T
authors.lastname: Kawano
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oikawa
authors.forename: Nobuo
authors.initials: N
authors.lastname: Kato
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sassa
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000737:chemistry / Q000378:metabolism; D001203:Ascomycota / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D003001:Cloning, Molecular; D004122:Dimethylallyltranstransferase / Q000378:metabolism; D004224:Diterpenes / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D005800:Genes, Fungal; D016681:Genome, Fungal / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D005810:Multigene Family; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA
chemical_list: D004224:Diterpenes; C539730:methyl phomopsenonate; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase; D004122:Dimethylallyltranstransferase