title: A fungal prenyltransferase catalyzes the regular di-prenylation at positions 20 and 21 of paxilline.
abstract: A putative indole diterpene biosynthetic gene cluster composed of eight genes was identified in a genome database of Phomopsis amygdali, and from it, biosynthetic genes of fusicoccin A were cloned and characterized. The six genes showed significant similarities to pax genes, which are essential to paxilline biosynthesis in Penicillium paxilli. Recombinants of the three putative prenyltransferase genes in the cluster were overexpressed in Escherichia coli and characterized by means of in vitro experiments. AmyG is perhaps a GGDP synthase. AmyC and AmyD were confirmed to be prenyltransferases catalyzing the transfer of GGDP to IGP and a regular di-prenylation at positions 20 and 21 of paxilline, respectively. AmyD is the first know example of an enzyme with this function. The Km values for AmyD were calculated to be 7.6 ± 0.5 μM for paxilline and 17.9 ± 1.7 μM for DMAPP at a kcat of 0.12 ± 0.003/s.
authors.affiliation: a Graduate School of Engineering , Hokkaido University , Sapporo , Japan.
authors.forename: Chengwei
authors.initials: C
authors.lastname: Liu
authors.forename: Motoyoshi
authors.initials: M
authors.lastname: Noike
authors.forename: Atsushi
authors.initials: A
authors.lastname: Minami
authors.forename: Hideaki
authors.initials: H
authors.lastname: Oikawa
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
mesh_terms: D001203:Ascomycota / Q000235:genetics; D003001:Cloning, Molecular; D004122:Dimethylallyltranstransferase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli; D051231:Farnesyltranstransferase / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D016681:Genome, Fungal; D007211:Indoles / Q000378:metabolism*; D005810:Multigene Family; D010407:Penicillium / Q000235:genetics; D054876:Prenylation
chemical_list: D007211:Indoles; C048220:paxilline; D004122:Dimethylallyltranstransferase; D051231:Farnesyltranstransferase
keywords: Phomopsis amygdali; fungi; indole diterpene; paxilline; prenyltransferases