Biosynthetic gene-based secondary metabolite screening: a new diterpene, methyl phomopsenonate, from the fungus Phomopsis amygdali.
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.
Toyomasu
Tomonobu
T
Department of Bioresource Engineering, Yamagata University, Tsuruoka, Yamagata 997-8555, Japan.
Kaneko
Akane
A
Tokiwano
Tetsuo
T
Kanno
Yuya
Y
Kanno
Yuri
Y
Niida
Rie
R
Miura
Shigeyoshi
S
Nishioka
Taiki
T
Ikeda
Chiho
C
Mitsuhashi
Wataru
W
Dairi
Tohru
T
Kawano
Tomikazu
T
Oikawa
Hideaki
H
Kato
Nobuo
N
Sassa
Takeshi
T
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
D004224:Diterpenes; C539730:methyl phomopsenonate; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase; D004122:Dimethylallyltranstransferase