L-Galactono-γ-lactone Dehydrogenase from Sweet Potato : Purification and cDNA Sequence Analysis
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概要
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L-Galactono-γ-lactone dehydrogenase(EC 1.3.2.3, GLDHase) was partially purified from mitochondria of sweet potato tuberous roots over 600-fold on a specific activity basis, followed by purification of the enzyme protein of 56 kDa by a preparative SDS-PAGE. The absorption spectrum of the hydroxylapatite column-purfied GLDHase showed peaks at 448 and 373 nm, suggesting the presence of flavin as a prosthetic group. The activity of GLDHase was inhibited by lycorine, an alkaloid which inhibits ascorbic acid biosynthesis in vivo. N-terminal partial sequences of four internal polypeptides generated by partial digestion of GLDHase with V8 protease were determined. The deduced nucleotide sequences were used to amplify a cDNA fragment of the GLDHase gene. The clone encoded a polypeptide of 581 amino acid residues with a molecular mass of 66 kDa. The deduced amino acid sequence showed 77% identity with that of cauliflower GLDHase, and significant homology to those of L-gulono-γ-lactone oxidase(22% identity) from rat and L-galactono-γ-lactone oxidase from yeast(17% identity), which are enzymes involved in L-ascorbic acid biosynthesis in these organisms. The absorption spectrum and cDNA sequence suggested that the flavin group bound noncovalently. We conclude that GLDHase, L-gulono-γ-lactone oxidase and L-galactono-γ-lactone oxidase are homologous in spite of the difference in substrates and electron acceptors. Genomic Southern analysis suggested that GLDHase gene exists as a single copy in the genome of sweet potato.
- 日本植物生理学会の論文
著者
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OBA Kazuko
Department of Food and Nutritional Sciences, College of Bioscience and Biotechnology, Chubu Universi
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Oba K
Nagoya Women's Univ. Nagoya Jpn
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Oba Kazuko
Dept. Food Sci. Nutr. Nagoya Women's Univ.
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Oba Kazuko
Laboratory Of Food Biochemistry Nagoya Women's University
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Oba Kazuko
Department Of Food And Nutritional Sciences College Of Bioscience And Biotechnology Chubu University
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KARITA Shuichi
Center for Molecular Biology and Genetics Mie University
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Hirai M
National Res. Inst. Vegetables Ornamental Plants And Tea Mie Jpn
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Hirai Masashi
Natl. Res. Inst. Veg. Ornam Plant & Tea.
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Nunome T
National Inst. Vegetable And Tea Sci. Mie Jpn
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Nunome Tsukasa
Natl. Res. Inst. Veg. Ornam Plant & Tea.
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Nunome Tsukasa
Molecular Genetics And Physiology Research Team National Institute Of Vegetable And Tea Science (niv
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Nunome Tsukasa
National Institute Of Vegetable And Tea Science National Agriculture And Food Research Organization
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Nunome Tsukasa
National Institute Of Vegetable And Tea Science (nivts)
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IMAI Tsuyoshi
Department of Electrical Engineering, Faculty of Engineering Science, Osaka University
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Imai Tuyoshi
Natl. Res. Inst. Veg. Ornam. Plant & Tea.
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Imai Tsuyoshi
Department Of Electrical Engineering Faculty Of Engineering Science Osaka University:tokai Research
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Karita S
Graduate School Of Regional Innovation Studies Mie Univ.
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Karita Shuichi
Graduate School Of Bioresources Mie Univ
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Shiratori Gen-ichi
Department of Vegetable Breeding, National Research Institute of Vegetables, Ornamental Plants and T
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Hattori Miyo
Department of Food Science and Nutrition, Nagoya Women's University
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Nunome Tsukasa
Department of Vegetable Breeding, National Research Institute of Vegetables, Ornamental Plants and T
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Hirai Masashi
Department of Vegetable Breeding, National Research Institute of Vegetables, Ornamental Plants and T
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Hattori Miyo
Dept. Food Sci. Nutr. Nagoya Women's Univ.
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Shiratori Gen-ichi
Natl. Res. Inst. Veg. Orn. Plants & Tea
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Hirai Masashi
Department Of Vegetable Breeding National Research Institute Of Vegetables Ornamental Plants & T
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Imai Tsuyoshi
Department Of Civil Engineering Yamaguchi Univerisity
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Hirai Masashi
Department Of Geosciences Osaka City University
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