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Identification of cDNAs encoding pterocarpan reductase involved in isoflavan phytoalexin biosynthesis in Lotus japonicus by EST mining
Authors:Akashi Tomoyoshi  Koshimizu Shoko  Aoki Toshio  Ayabe Shin-Ichi
Institution:Department of Applied Biological Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-8510, Japan. akashi@brs.nihon-u.ac.jp
Abstract:Isoflavans and pterocarpans are the major biosynthetically connected phytoalexins in legumes. A search of the expressed sequence tag library of a model legume Lotus japonicus, which produces an (-)-isoflavan, for homologs of phenylcoumaran benzylic ether reductase catalyzing the reductive cleavage of dihydrofurans, yielded seven full-length cDNAs, and the encoded proteins were analyzed in vitro. Four of them cleaved the dihydrofuran of a pterocarpan medicarpin to yield an isoflavan (-)-vestitol and were designated pterocarpan reductase (PTR). Two PTRs displayed enantiospecificity to (-)-medicarpin, representing genuine L. japonicus PTRs, while the other two lacked enantiospecificity and were presumed to be evolutionarily primitive types.
Keywords:DMID  7  2′-dihydroxy-4′-methoxyisoflavanol dehydratase  EST  expressed sequence tag  GSH  reduced glutathione  IFR  isoflavone reductase  NMR  nuclear magnetic resonance  PCBER  phenylcoumaran benzylic ether reductase  PLR  pinoresinol-lariciresinol reductase  PTR  pterocarpan reductase  Rt  retention time  SDR  short-chain dehydrogenase/reductase  VR  vestitone reductase
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