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
- 来源:
- 关键词:
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.Mori, A.Tatematsu, A.Saito, H.Matsui, A.Kimura & S.Chiba: "Substrate Specificity and Subsite Affinties of alpha-Amylase from Germinating Cotyledons of Phaseolus vulgaris L.ev Toramame" Oyo Toshitsu Kagaku (J.Appl.Glycosci.). 42 (4). 387-394 (1995)
H.Mori、A.Tatematsu、A.Saito、H.Matsui、A.Kimura
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- 影响因子:0
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H.Kawagishi, T.Wasa, T.Murata, T.Usui, A.Kimura & S.Chiba: "Two N-acetyl-D-galactosamine-specific Lectins, PAL-1 and PAL-11, from the Mushroom Phaeolepiota aurea" Phytochemistry. 41 (4). 1013-1016 (1996)
H.川岸、T.Wasa、T.Murata、T.Usui、A.Kimura
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- 影响因子:0
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R.Surarit, H.Matsui, S.Chiba, J.Svasti & C.Srisomsap: "Chemical Modification of beta-Glucosidase/beta-Fucosidase from Dalbergia Cochinchinensis Pierre by Conduritol B Epoxide" Biosci.Biotech.Biochem.60 (8). 1265-1268 (1996)
R.Surarit、H.Matsui、S.Chiba、J.Svasti
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- 影响因子:0
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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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- 影响因子:0
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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)