| RRC ID |
50640
|
| 著者 |
Iijima M, Munakata R, Takahashi H, Kenmoku H, Nakagawa R, Kodama T, Asakawa Y, Abe I, Yazaki K, Kurosaki F, Taura F.
|
| タイトル |
Identification and Characterization of Daurichromenic Acid Synthase Active in Anti-HIV Biosynthesis.
|
| ジャーナル |
Plant Physiol
|
| Abstract |
Daurichromenic acid (DCA) synthase catalyzes the oxidative cyclization of grifolic acid to produce DCA, an anti-HIV meroterpenoid isolated from Rhododendron dauricum We identified a novel cDNA encoding DCA synthase by transcriptome-based screening from young leaves of R. dauricum The gene coded for a 533-amino acid polypeptide with moderate homologies to flavin adenine dinucleotide oxidases from other plants. The primary structure contained an amino-terminal signal peptide and conserved amino acid residues to form bicovalent linkage to the flavin adenine dinucleotide isoalloxazine ring at histidine-112 and cysteine-175. In addition, the recombinant DCA synthase, purified from the culture supernatant of transgenic Pichia pastoris, exhibited structural and functional properties as a flavoprotein. The reaction mechanism of DCA synthase characterized herein partly shares a similarity with those of cannabinoid synthases from Cannabis sativa, whereas DCA synthase catalyzes a novel cyclization reaction of the farnesyl moiety of a meroterpenoid natural product of plant origin. Moreover, in this study, we present evidence that DCA is biosynthesized and accumulated specifically in the glandular scales, on the surface of R. dauricum plants, based on various analytical studies at the chemical, biochemical, and molecular levels. The extracellular localization of DCA also was confirmed by a confocal microscopic analysis of its autofluorescence. These data highlight the unique feature of DCA: the final step of biosynthesis is completed in apoplastic space, and it is highly accumulated outside the scale cells.
|
| 巻・号 |
174(4)
|
| ページ |
2213-2230
|
| 公開日 |
2017-8-1
|
| DOI |
10.1104/pp.17.00586
|
| PII |
pp.17.00586
|
| PMID |
28679557
|
| PMC |
PMC5543965
|
| MeSH |
Anti-HIV Agents / metabolism*
Biocatalysis
Biosynthetic Pathways*
Chromans / metabolism*
Chromatography, High Pressure Liquid
Cloning, Molecular
Electrophoresis, Polyacrylamide Gel
Gene Expression Regulation, Plant
Green Fluorescent Proteins / metabolism
Kinetics
Ligases / genetics
Ligases / metabolism*
Oxygen / metabolism
Phylogeny
Phytochemicals / metabolism
Pichia / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
Recombinant Proteins / metabolism
Rhododendron / cytology
Rhododendron / genetics
Rhododendron / metabolism
Structural Homology, Protein
Tobacco / cytology
|
| IF |
6.902
|
| 引用数 |
8
|
| リソース情報 |
| シロイヌナズナ / 植物培養細胞・遺伝子 |
rpc00001 |
