title: Comparative functional analysis of CYP71AV1 natural variants reveals an important residue for the successive oxidation of amorpha-4,11-diene.
abstract: Artemisinin is an antimalarial sesquiterpenoid isolated from the aerial parts of the plant Artemisia annua. CYP71AV1, a cytochrome P450 monooxygenase was identified in the artemisinin biosynthetic pathway. CYP71AV1 catalyzes three successive oxidation steps at the C12 position of amorpha-4,11-diene to produce artemisinic acid. In this study, we isolated putative CYP71AV1 orthologs in different species of Artemisia. Comparative functional analysis of CYP71AV1 and its putative orthologs, together with homology modeling, enabled us to identify an amino acid residue (Ser479) critical for the second oxidation reaction catalyzed by CYP71AV1. Our results clearly show that a comparative study of natural variants is useful to investigate the structure-function relationships of CYP71AV1.
authors.affiliation: Kihara Institute for Biological Research, Yokohama City University, Yokahama, Kanagawa 244-0813, Japan.
authors.forename: Aya
authors.initials: A
authors.lastname: Komori
authors.forename: Munenori
authors.initials: M
authors.lastname: Suzuki
authors.forename: Hikaru
authors.initials: H
authors.lastname: Seki
authors.forename: Tomoko
authors.initials: T
authors.lastname: Nishizawa
authors.forename: Jacobus Johannes Marion
authors.initials: JJ
authors.lastname: Meyer
authors.forename: Hideaki
authors.initials: H
authors.lastname: Shimizu
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Toshiya
authors.initials: T
authors.lastname: Muranaka
mesh_terms: D000595:Amino Acid Sequence; D031171:Artemisia absinthium / Q000201:enzymology / Q000235:genetics; D031172:Artemisia annua / Q000201:enzymology / Q000235:genetics; D055162:Biocatalysis; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010084:Oxidation-Reduction; D000081223:Polycyclic Sesquiterpenes; D011487:Protein Conformation; D012689:Sequence Homology, Nucleic Acid; D012717:Sesquiterpenes / Q000378:metabolism*; D013045:Species Specificity
chemical_list: D000081223:Polycyclic Sesquiterpenes; D012717:Sesquiterpenes; C515348:amorpha-4,11-diene; D003577:Cytochrome P-450 Enzyme System