title: Biosynthetic conclusions from the functional dissection of oxygenases for biosynthesis of actinorhodin and related Streptomyces antibiotics.
abstract: Actinorhodin (ACT) produced by Streptomyces coelicolor A3(2) belongs to the benzoisochromanequinone (BIQ) class of antibiotics. ActVA-ORF5, a flavin-dependent monooxygenase (FMO) essential for ACT biosynthesis, forms a two-component enzyme system in combination with a flavin:NADH oxidoreductase, ActVB. The genes for homologous two-component FMOs are found in the biosynthetic gene clusters for two other BIQs, granaticin (GRA) and medermycin (MED), and a closely related antibiotic, alnumycin (ALN). Our functional analysis of these FMOs (ActVA-ORF5, Gra-ORF21, Med-ORF7, and AlnT) in S. coelicolor unambiguously demonstrated that ActVA-ORF5 and Gra-ORF21 are bifunctional and capable of both p-quinone formation at C-6 in the central ring and C-8 hydroxylation in the lateral ring, whereas Med-ORF7 catalyzes only p-quinone formation. No p-quinone formation on a BIQ substrate was observed for AlnT, which is involved in lateral p-quinone formation in ALN.
authors.affiliation: Research Institute of Pharmaceutical Sciences, Musashino University, Shinmachi, Nishitokyo-shi, Tokyo 202-8585, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Taguchi
authors.forename: Masaki
authors.initials: M
authors.lastname: Yabe
authors.forename: Hitomi
authors.initials: H
authors.lastname: Odaki
authors.forename: Miki
authors.initials: M
authors.lastname: Shinozaki
authors.forename: Mikko
authors.initials: M
authors.lastname: Metsä-Ketelä
authors.forename: Takao
authors.initials: T
authors.lastname: Arai
authors.forename: Susumu
authors.initials: S
authors.lastname: Okamoto
authors.forename: Koji
authors.initials: K
authors.lastname: Ichinose
mesh_terms: D000880:Anthraquinones / Q000737:chemistry / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism; D009285:Naphthoquinones / Q000378:metabolism; D011809:Quinones / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D021241:Spectrometry, Mass, Electrospray Ionization; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics; D000095622:Benzoisochromanequinones
chemical_list: D000880:Anthraquinones; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D006899:Mixed Function Oxygenases; D009247:NADH, NADPH Oxidoreductases; D009285:Naphthoquinones; D011809:Quinones; D011994:Recombinant Proteins; C112189:alnumycin; C009811:granaticin; D000095622:Benzoisochromanequinones; C013390:actinorhodin