Analyzes for Mechanism of Substate Recognition and Structure of Sugar Chain in Fungus Crystalline alpha-Glucosidase

真菌结晶α-葡萄糖苷酶的物质识别机制和糖链结构分析

基本信息

  • 批准号:
    06453164
  • 负责人:
  • 金额:
    $ 4.8万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
  • 财政年份:
    1994
  • 资助国家:
    日本
  • 起止时间:
    1994 至 1996
  • 项目状态:
    已结题

项目摘要

An crystalline alpha-glucosidase from Asperillus niger (ANGase), a so-called "maltase"-type enzyme, was composed of two different subunits, p1 (227 amino acids) and P2 (719 amino acids), which were not linked via S-S bridge. Their primary structures were determined by Edman degradation and genetic methods. The genomic nucleotide sequence revealed theat the P1 and P2 polypeptides were encoded by a single reading frame in this order. Between two subunits 14 amino acids were removed, suggesting an unique post-translational modification with limited proteolysis. The Asp (224) of P2 was identified to be one of catalytic amino acids. Seven N-linked high-mannose type oligosaccharides were isolated, and their structures were determined. Four sugar chains contained one galactofuranosyl residue bound via alpha- (1*2) -linkage to Man (A). Three of them are novel types of N-linked oligosaccharides. The O-linked sugars also were isolated from ANGase. Structural analysis of two compounds indicated that mannose and alpha-mannosyl- (1*2) -mannose bound directly to Ser/Thr of ANGase.Based on the above findings, A.niger alpha-glucosidase was compared in the primary structures with alpha-glucosidases from other origins. alpha-Glucosidases were elucidated to be classified into the two families on the primary structures. In adittion, the mode of substrate recognition and the model of reaction mechanism in the carbohydrate-hydrolases were discussed and proposed on the analysis for the hydration reaction of D-glucal and the alpha-secondary kinetic isotope effects by alpha-glucosidase and glucoamylase.
来自尼日尔曲霉的结晶α-葡萄糖苷酶(ANGase)是一种所谓的“麦芽糖酶”型酶,由两个不同的亚基P1(227个氨基酸)和P2(719个氨基酸)组成,它们不通过S-S桥连接。通过Edman降解和遗传学方法确定了它们的一级结构。基因组核苷酸序列分析表明,P1和P2多肽由一个阅读框依次编码。两个亚基之间的14个氨基酸被删除,这表明一个独特的翻译后修饰有限的蛋白水解。P2的Asp(224)被鉴定为催化氨基酸之一。分离得到7个N-连接高甘露糖型寡糖,并测定了它们的结构。四条糖链含有一个通过α-(1*2)-键与Man结合的呋喃半乳糖残基(A)。其中三种是新型的N-连接寡糖。O-连接的糖也从ANGase分离。对两个化合物的结构分析表明,甘露糖和α-甘露糖基-(1*2)-甘露糖直接与ANGase的Ser/Thr结合,在此基础上,将黑尼日尔α-葡萄糖苷酶与其他来源的α-葡萄糖苷酶进行了一级结构的比较。α-葡萄糖苷酶在一级结构上可分为两个家族。同时,在分析D-葡萄糖醛水合反应以及α-葡萄糖苷酶和葡萄糖淀粉酶的α-二级动力学同位素效应的基础上,讨论并提出了碳水化合物水解酶的底物识别方式和反应机理模型。

项目成果

期刊论文数量(87)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
H.Matsui, S.Iwanami, H.Ito, A.Kimura, H.Mori, M.Honma and S.Chiba: "Substrate Specificity and Primary Structure of Sugar Beet alpha-Glucosidase" Oyo Toshitsu Kagaku. 44 (in press). (1997)
H.Matsui、S.Iwanami、H.Ito、A.Kimura、H.Mori、M.Honma 和 S.Chiba:“甜菜 α-葡萄糖苷酶的底物特异性和一级结构”Oyo Toshitsu Kagaku。
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    0
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  • 通讯作者:
S.Chiba: "Molecular Mechanism of Glycosidases-Namely alpha-Glucosidase-in "New Approach for Enzyme Studies" (ed.by E.Ichishima and S.Onodera)" Tokyo Kagaku Dojin Inc.70-82 (1995)
S.Chiba:“酶研究的新方法”中糖苷酶(即α-葡萄糖苷酶)的分子机制(E.Ichishima 和 S.Onodera 编)”东京化学同人株式会社.70-82(1995)
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CHIBA Seiya其他文献

CHIBA Seiya的其他文献

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{{ truncateString('CHIBA Seiya', 18)}}的其他基金

Joint Studies on Dextran-Producing and -Degrading Enzymes
右旋糖酐生产和降解酶的联合研究
  • 批准号:
    10044190
  • 财政年份:
    1998
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Structures and Substrate Recognitions of Animal, Botanical and Microbial α-Glucosidases and Their Molecular Evolution
动物、植物和微生物 α-葡萄糖苷酶的结构和底物识别及其分子进化
  • 批准号:
    09460041
  • 财政年份:
    1997
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
New Technique for High Yield Production of Glucose by Using of Mutarotase
变旋酶高产生产葡萄糖新技术
  • 批准号:
    05556011
  • 财政年份:
    1993
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research (B)
Studies on Mechanism of Substrate Confirmation, Modification of Function, and Utilization for Carbohydrate-Hydrolase and Synthase
碳水化合物水解酶和合酶的底物确认、功能修饰和利用机制研究
  • 批准号:
    05303010
  • 财政年份:
    1993
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for Co-operative Research (A)
Structure and Expression of Mold Crystalline alpha-Glucosidase
霉菌结晶α-葡萄糖苷酶的结构和表达
  • 批准号:
    04453129
  • 财政年份:
    1992
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
Kinetic Studies on Allosteric Properties of Honeybee alpha-Glucosidase
蜜蜂α-葡萄糖苷酶变构特性的动力学研究
  • 批准号:
    02660074
  • 财政年份:
    1990
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
New Method Using Bio-Reactor for Production of Isomaltose as Food Stuff
利用生物反应器生产食品异麦芽糖的新方法
  • 批准号:
    01860012
  • 财政年份:
    1989
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research
Mechanism of Occurrence of Carbohydrate Metabolizing Enzymes in Corn Seeds.
玉米种子中碳水化合物代谢酶的发生机制。
  • 批准号:
    62560067
  • 财政年份:
    1987
  • 资助金额:
    $ 4.8万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
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