An Active-Site Mutation Causes Enhanced Reactivity and Altered Regiospecificity of Transglucosylation Catalyzed by the Bacillus sp.SAM1606 α-Glucosidase

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概要

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• Bacillus sp.SAM1606 α-glucosidase catalyzes the transglucosylation of sucrose to produce three regioisomers of the glucosylsucroses, with theanderose(6-O^G-glucosylsucrose) as the most abundant transfer product. To find the active-site amino acid residues with can affect the reactivity and regiospecificity of the glucosyl transfer, 16 mutants with amino acid substitutions near the active site were allowed to react with 1.75 M sucrose at 60℃, pH6.0, and the course of transglucosylation as well as the product specificity were analyzed. The sites of the amino acid substitutions were selected by comparing the conserved amino acid sequences located near the active site of the SAM1606 enzyme with those of the Bacillus oligo-1, 6-glucosidases(O16G), which have very high amino acid sequence similarities near the active site but have a distinct substrate specificity. The results showed that, among the mutated SAM1606 enzymes examined, only the mutants with substitution of Gly273 with Pro showed an altered reactivity and specificity of transglucosylation; these mutants exhibited a significantly enhanced initial velocity of glucosyl transfer, yielding isomelezitose(6-O^F-glucosylsucrose) instead of theanderose as the major transfer product. These results indicate that the substitution of Gly273 with Procritically governs the enhanced reactivity and altered specificity of the transglucosylation. The notion that the amino acid residue at this position is the determinant of the glucosyl-transfer specificity was further confirmed by observation that the Bacillus cereus O16G, which has a proline at the corresponding position, produced isomelezitose as the major transfer product during transglucosylation with sucrose.

• 社団法人日本生物工学会の論文
• 2000-05-25

著者

• Nishino Tokuzo

  Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University

• Nakayama Toru

  Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University

• 石井 正浩

  University Of Tokyo

• Nishino T

  Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University

• 石井 正治

  東京大学大学院農学生命科学研究科応用工学専攻

• UEDA Takashi

  Department of Surgery, National Hospital Organization Saga Hospital

• Nishino Tokuzo

  Dep. Of Biochemistry And Engineering Graduate School Of Engineering Tohoku Univ. Aoba-yama Sendai Mi

• Ishii M

  Department Of Biotechnology Graduate School Of Agricultural And Life Sciences University Of Tokyo

• Ishii Masaharu

  Dep. Of Biotechnology Graduate School Of Agricultural And Life Sciences Univ. Of Tokyo

• Hemmi H

  Tohoku Univ. Miyagi Jpn

• Hemmi Hisashi

  Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University

• Hemmi Hisashi

  Department Of Biochemistry And Engineering Tohoku University

• Hemmi Hisashi

  Dep. Of Biochemistry And Engineering Graduate School Of Engineering Tohoku Univ. Aoba-yama Sendai Mi

• OKADA Maki

  Institute for Advanced Technology, Suntory Ltd.

• INOHARA-OCHIAI MISA

  Suntory Research Center

• OKADA MAKI

  Department of Nutritional Physiology, Faculty of Nutrition, Kobe Gakuin University

• IWASHITA TAKASHI

  Suntory Institute for Bioorganic Research(SUNBOR)

• KAN YUKIKO

  Suntory Institute for Bioorganic Research(SUNBOR)

• SHIBANO YUJI

  Department of Nutritional Physiology, Faculty of Nutrition, Kobe Gakuin University

• ASHIKARI TOSHIHIKO

  Department of Nutritional Physiology, Faculty of Nutrition, Kobe Gakuin University

• Nakayama Toru

  Department Of Biomolecular Engineering Graduate School Of Engineering Tohoku University

• Iwashita T

  Suntory Institute For Bioorganic Research

• Iwashita Takashi

  Suntory Inst. For Bioorganic Res. Osaka Jpn

• Shibano Yuji

  Suntory Research Center:(present Address)suntory Ltd.

• Shibano Yuji

  Department Of Molecular Biology Biomolecular Engineering Research Institute

• Shibano Yuji

  Department Of Moleular Biology Biomolecular Engineering Research Insitute

• Amachi T

  Kyoto Univ. Kyoto Jpn

• Hamada Hiroaki

  Department Of Life Science Faculty Of Science Okayama University Of Science

• Nishino Tokuzo

  Department Of Biochemistry And Engineering Faculty Of Engineering Tohoku University

• Ishii Masaharu

  Graduate School Of Agricultural And Life Sciences The University Of Tokyo

• Okada Mitsumasa

  Institute For Advanced Technology Suntory Ltd.

• Kan Yukiko

  Suntory Institute For Bioorganic Research (sunbor)

• Ueda Takashi

  Department Of Biological Sciences Graduate School Of Science The University Of Tokyo

• Hemmi Hisashi

  Department Of Applied Molecular Biosciences Graduate School Of Bioagricultural Sciences Nagoya Unive

• Ueda T

  Department Of Nutritional Physiology Faculty Of Nutrition Kobe Gakuin University

• Ueda Takashi

  Department Of Biochemistry And Molecular Biology Kyoto Pharmaceutical University

• Yamashita Satoshi

  Department Of Biochemistry And Engineering Graduate School Of Engineering Tohoku University

• Ueda Takashi

  Department Of Applied Biology And Chemistry Tokyo University Of Agriculture

• Okada Maki

  Department Of Neurosurgery Shinshu University School Of Medicine

• Kan Yukiko

  Suntory Institute for Bioorganic Research

• NAKAYAMA Toru

  Department of Applied Microbial Technology, The Kumamoto Institute of Technology:(Present office)Institute for Fundamental Research, Suntory Ltd.

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