Studies on dissolved metalloenzymes in lake water. I. Identification of alkaline phosphatase.
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概要
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Metalloenzyme (alkaline phosphatase) dissolved in lake water has been identified by means of high performance liquid chromatography (HPLC). Water samples were collected from Lake Kasumigaura in Ibaraki, and filtered with a 0.2–0.45 μm membrane filter immediately after sampling. Filtered water was concentrated to 100–2000 fold by ultrafiltration membrane which could concentrate the components of molecular weight larger than 10000. The enzymatic activity of alkaline phosphatase was measured using <I>p</I>-nitrophenyl dihydrogen-phosphate as a substrate. Chelating agents added to the concentrated lake water inhibited the appearance of enzymatic activity, while the enzymatic activity was recovered with addition of zinc ion. The HPLC chromatograms of the concentrated lake water were examined, where an aqueous porous gel column was used for separation. The effluent at molecular weight of <I>ca.</I> 90000 showed specific activity of alkaline phosphatase. Zinc was also found in the same fraction. These facts suggest that alkaline phosphatase with the nature similar to procaryotes was present in the lake water.
- 公益社団法人 日本化学会の論文
著者
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Kobayashi Kensei
Department Of Chemistry And Biotechnology Faculty Of Engineering Yokohama National University
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Fujiwara Kitao
Department Of Chemistry Faculty Of Science University Of Tokyo
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Fuwa Keiichiro
Department Of Chemistry Faculty Of Science University Of Tokyo
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Matsui Masahiro
Department Of Architecture Tokyo Polytechnic University
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Watanabe Masami
Department Of Perinatology Institute For Developmental Research
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Haraguchi Hiroki
Department of Agricultural Chemistry, Faculty of Agriculture The University of Tokyo
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Matsui Masahiro
Department of Chemistry, Faculty of Science, The University of Tokyo
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Iwase Kimikazu
Department of Chemistry, Faculty of Science, The University of Tokyo
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Ueda Hirotaka
Department of Chemistry, Faculty of Science, The University of Tokyo
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Watanabe Masami
Department of Applied Chemistry, Faculty of Science, Science University of Tokyo
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Fuwa Keiichiro
Department of Chemistry and Physics, National Institute for Environmental Studies
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Fuwa Keiichiro
Department of Chemistry, Faculty of Science, The University of Tokyo
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Fuwa Keiichiro
Department of Agricultural Chemistry, Faculty of Agriculture The University of Tokyo
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