Purification and Characterization of Glucosyltransferase and Glucanotransferase Involved in the Production of Cyclic Tetrasaccharide in Bacillus globisporus C11(Biochemistry & Molecular Biology)
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概要
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Glucosyltransferase and glucanotransferase involved in the production of cyclic tetrasaccharide (CTS ; cyclo{→6}-α-D-glucopyranosyl-(1→3)-α-D-glucopyranosyl-(1→6)-α-D-glucopyranosyl-(1→3)-α-D-glucopyranosyl-(1→)) from α-1, 4-glucan were purified from Bacillus globisporus C11. The former was a 1, 6-α-glucosyltransferase (6GT) catalyzing the α-1, 6-transglucosylation of one glucosyl residue to the nonreducing end of maltooligosaccharides (MOS) to produce α-isomaltosyl-MOS from MOS. The latter was an isomaltosyl transferase (IMT) catalyzing α-1, 3-, α-1, 4-, and α, β-1, 1-intermolecular transglycosylation of isomaltosyl residues. When IMT catalyzed α-1, 3-transglycosylation, α-isomaltosyl-(1→3)-α-isomaltosyl-MOS was produced from α-isomaltosyl-MOS. In addition, IMT catalyzed cyclization, and produced CTS from α-isomaltosyl-(1→3)-α-isomaltosyl-MOS by intramolecular transglycosylation. Therefore, the mechanism of CTS synthesis from MOS by the two enzymes seemed to follow three steps : 1) MOS→α-isomaltosyl-MOS (by 6GT), 2) α-isomaltosyl-MOS→α-isomaltosyl-(1→3)-α-isomaltosyl-MOS (by IMT), and 3) α-isomaltosyl-(1→3)-α-isomaltosyl-MOS→CTS+MOS (by IMT). The molecular mass of 6GT was estimated to be 137 kDa by SDS-PAGE. The optimum pH and temperature for 6GT were pH 6.0 and 45℃, respectively. This enzyme was stable at from pH 5.5 to 10 and on being heated to 40℃ for 60 min. 6GT was strongly activated and stabilized by various divalent cations. The molecular mass of IMT was estimated to be 102 kDa by SDS-PAGE. The optimum pH and temperature for IMT were pH 6.0 and 50℃, respectively. This enzyme was stable at from pH 4.5 to 9.0 and on being heated to 40℃ for 60 min. Divalent cations had no effect on the stability or activity of this enzyme.
- 社団法人日本農芸化学会の論文
- 2002-09-23
著者
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NISHIMOTO Tomoyuki
Hayashibara Biochemical Laboratories Inc.
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Fukuda S
Biomedical Inst. Res. Center Hayashibara Biochemical Laboratories Inc.
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Kurimoto M
Amase Institute Hayashibara Biochemical Laboratories Inc.
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Kurimoto Masashi
Hayashibara Biochemical Laboratories Inc. Fujisaki Institute
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Kurimoto Masashi
Hayashibara Biochemical Laboratories
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Watanabe H
林原生物化学研究所
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Watanabe Hikaru
Glycoscience Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Aga H
Glycoscience Institute Hayashibara Biochemical Laboratories Inc.
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FUKUDA Shigeharu
Hayashibara Biochemical Laboratories
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TSUJISAKA Yoshio
Hayashibara Institute Corp.
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NISHIMOTO Tomoyuki
Glycoscience Institute, Research Center, Hayashibara Biochemical Laboratories, Inc.
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KUBOTA Michio
Hayashibara Biochemical Laboratories, Inc.
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Taniguchi Yoshifumi
Biomedical Institute, Research Center, Hayashibara Biochemical Laboratories, Inc.
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Kubota Michio
Hayashibara Biochemical Laboratories Inc.
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Kurose Mayumi
Glycoscience Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Mukai Kazuhisa
Glycoscience Institute Research Center Hayashibara Biochemical Laboratories Inc.
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HIGASHIYAMA TAKANOBU
Amase Institute, Hayashibara Biochemical Laboratories, Inc.
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AGA Hajime
Hayashibara Biochemical Laboratories, Inc.
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WATANABE Hikaru
Hayashibara Biochemical Laboratories, Inc.
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MUKAI Kazuhisa
Hayashibara Biochemical Laboratories, Inc.
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HASHIMOTO Takaharu
Hayashibara Biochemical Laboratories, Inc.
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Tsujisaka Y
Biomedical Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Nishimoto Tomoyuki
Glycoscience Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Maruta K
Glycoscience Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Taniguchi Yoshifumi
Biomedical Institute Research Center Hayashibara Biochemical Laboratories Inc.
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Higashiyama Takanobu
Amase Institute Hayashibara Biochemical Laboratories Inc.
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