Molecular Cloning, Functional Expression, and Tissue Distribution of a Potato Sprout Allene Oxide Synthase Involved in a 9-Lipoxygenase Pathway

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概要

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• Potato (Solanum tuberosum) plants are rich in 9-lipoxygenase, which converts linoleic acid and α-linolenic acid to 9S-hydroperoxy-10E,12Z-octadecadienoic acid (9-HPOD) and 9S-hydroperoxy-10E,12Z,15Z-octadecatrienoic acid (9-HPOT) respectively. The allene oxide synthase (AOS) involved in 9-HPOD/9-HPOT metabolism in potato, however, has not been characterized in detail. We cloned a cDNA encoding a novel AOS from potato sprouts by reverse transcriptase-PCR based on a partial sequence in the EST database. This AOS was successfully expressed in the yeast Pichia pastoris, and purified using Ni-NTA resin. The recombinant enzyme metabolized 9-HPOD, 9-HPOT, 13-HPOD, and 13-HPOT with reaction efficiencies of 2.5×107, 1.0×107, 2.5×106, and 7.6×106 M−1 s−1 respectively. The α-ketol formed from 9-HPOD was composed mainly of the 9R-enatimomer (90%). Besides sprouts, the mRNA of this AOS was detected in buds, flowers, and stems, but not in leaves, tubers, or roots of mature plants, suggesting that this enzyme has a tissue-specific function.

• 2007-09-23

著者

• 森 春英

  北海道大学大学院農学研究科応用生命科学専攻

• 森 春英

  北海道大学大学院農学研究院応用生命科学部門

• Yokota K

  Department Of Bacteriology Okayama University Graduate School Of Medicine And Dentistry

• WONGCHAWALIT Jintanart

  Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University

• LEE Jin-Ha

  Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University

• Chiba Seiya

  Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University

• Nishimura Kohji

  Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Techn

• NISHIGAICHI Yutaka

  Department of Chemistry, Faculty of Science, Shimane University

• Kim Doman

  School Of Biological Sciences And Technology And Institute Of Bioindustrial Technology Jeonnam Natio

• Kobayasi Kazuhiro

  Department Of Agriculture Faculty Of Life And Environmental Science Shimane University

• Nagaya Tsutomu

  Department Of Life Science And Biotechnology Shimane University

• Kimura Atsuo

  Division Of Applied Bioscience Graduate School Of Agriculture Hokkaido University

• KONGRIT Darika

  Division of Applied Resources Chemistry, Course of Bioresources Science, The United Graduate School

• JISAKA Mitsuo

  Division of Applied Resources Chemistry, Course of Bioresources Science, The United Graduate School

• NAGAYA Tsutomu

  Division of Applied Resources Chemistry, Course of Bioresources Science, The United Graduate School

• YOKOTA Kazushige

  Division of Applied Resources Chemistry, Course of Bioresources Science, The United Graduate School

• Mori Haruhide

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Jisaka Mitsuo

  Division Of Applied Resources Chemistry The United Graduate School Of Agricultural Sciences Tottori

• Saito Saori

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Okuyama Masayuki

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Nishimoto Mamoru

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Kimura Atsuo

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Kongrit Darika

  Division Of Applied Resources Chemistry The United Graduate School Of Agricultural Sciences Tottori

• Kim Doman

  School Of Biological Sciences And Technology And Institute Of Bioindustrial Technology Jeonnam Natio

• Wongchawalit Jintanart

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Lee Jin-ha

  Division Of Applied Bioscience Research Faculty Of Agriculture Hokkaido University

• Mori Haruhide

  Research Faculty Of Agriculture Hokkaido University

• Kobayasi Kazuhiro

  Division of Applied Bioscience, Research Faculty of Agriculture, Hokkaido University

• Kim Doman

  School of Biological Science and Biotechnology, Chonnam National University

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