title: Reveromycin A biosynthesis uses RevG and RevJ for stereospecific spiroacetal formation.
abstract: Spiroacetal compounds are ubiquitous in nature, and their stereospecific structures are responsible for diverse pharmaceutical activities. Elucidation of the biosynthetic mechanisms that are involved in spiroacetal formation will open the door to efficient generation of stereospecific structures that are otherwise hard to synthesize chemically. However, the biosynthesis of these compounds is poorly understood, owing to difficulties in identifying the responsible enzymes and analyzing unstable intermediates. Here we comprehensively describe the spiroacetal formation involved in the biosynthesis of reveromycin A, which inhibits bone resorption and bone metastases of tumor cells by inducing apoptosis in osteoclasts. We performed gene disruption, systematic metabolite analysis, feeding of labeled precursors and conversion studies with recombinant enzymes. We identified two key enzymes, dihydroxy ketone synthase and spiroacetal synthase, and showed in vitro reconstruction of the stereospecific spiroacetal structure from a stable acyclic precursor. Our findings provide insights into the creation of a variety of biologically active spiroacetal compounds for drug leads.
authors.affiliation: Chemical Biology Department, RIKEN Advanced Science Institute, Saitama, Japan.
authors.forename: Shunji
authors.initials: S
authors.lastname: Takahashi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Toyoda
authors.forename: Yasuyo
authors.initials: Y
authors.lastname: Sekiyama
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Takagi
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Nogawa
authors.forename: Masakazu
authors.initials: M
authors.lastname: Uramoto
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Suzuki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Koshino
authors.forename: Takuto
authors.initials: T
authors.lastname: Kumano
authors.forename: Suresh
authors.initials: S
authors.lastname: Panthee
authors.forename: Tohru
authors.initials: T
authors.lastname: Dairi
authors.forename: Jun
authors.initials: J
authors.lastname: Ishikawa
authors.forename: Haruo
authors.initials: H
authors.lastname: Ikeda
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Sakaki
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Osada
mesh_terms: D053898:Biosynthetic Pathways; D002851:Chromatography, High Pressure Liquid; D003500:Cyclization; D013058:Mass Spectrometry; D015394:Molecular Structure; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D011714:Pyrans / Q000737:chemistry*; D013141:Spiro Compounds / Q000737:chemistry*; D013237:Stereoisomerism; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics
chemical_list: D011714:Pyrans; D013141:Spiro Compounds; C068098:reveromycin A; D048630:Polyketide Synthases