Studies on glycosidases : Recognition of substrate and alteration of function

糖苷酶的研究：底物识别和功能改变

• 批准号: 11460032
• 负责人: MATUI Hirokazu
• 金额: $ 2.75万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11460032/
• 关键词: Glycosidase; Glucodextranase; Isomaltotrio-dextranase; Starch branching enzyme; Chimeric enzymes; Substnate Specificity; 枝作り酵素; デキストラナーゼ; α-グルコシダーゼ; グランチングエンザイム; glucodextranase(グルコデキストラナーゼ); isomaltotrio-dextranase(イソマルトトリオデキストラナーゼ); glucoamy

I cloned the glucodextranase gene, and found that showed the similarity to some of glucoamylase, it is possible to determine the region for bind to α-1,4 orα-1,6 glucosidic linkages. Also the isomaltotrio-dextranase gene was isolated and expressed in E. coll. The expressed enzyme was analyzed.Starch branching enzyme (BE) is concern to biosynthesis of starch, which catalyze the hydrolysis ofα-1,4-glucosidic linkage and transferring it to make α-1,6 point. I cloned two types of this enzyme, and succcessed to express them in E, coll. Chimeric enzymes from these isozymes were analyzed. N-terminal was important to determine the affinity to amylose or amylopectin. And some chimeric enzymes which have different properties were made and analyzed.

我克隆了葡聚糖酶基因，发现它与一些糖淀粉酶基因有相似性，可以确定与α-1，4或α-1，6糖苷键结合的区域。此外，还分离了异麦芽三糖葡聚糖酶基因，并在E.coll中进行了表达。淀粉分支酶(BE)与淀粉的生物合成有关，它催化α-1，4-葡萄糖苷键的水解，并将其转移到α-1，6点。我克隆了两种类型的这种酶，并成功地在E，coll中表达了它们。对这些同工酶的嵌合酶进行了分析。N-末端是决定与直链淀粉或支链淀粉亲和力的重要因素。并对不同性质的嵌合酶进行了制备和分析。

项目成果

M.Nishimoto: "Purification and substrate Specificity of Honeybee, Apis mellifera L, α-Glues : damsel"Bioscience, Biotechnology, and Biochemistry. 65. 1610-1616 (2001)

M. Nishimoto：“蜜蜂、Apis mellifera L、α-Glues 的纯化和底物特异性：damsel”《生物科学、生物技术和生物化学》65。1610-1616 (2001)。

M. Nishimoto, M. Kubota, M. Tsuji, H. Mori, A. Kimura, H. Matsui, S. Chiba: "Purification and Substrate Specificity of Honeybee, Apis mellifera L., α-Glucosidase III"Biosci, Biotech. Biochem.. 65. 1610-1616 (2001)

M. Nishimoto、M. Kubota、M. Tsuji、H. Mori、A. Kimura、H. Matsui、S. Chiba：“蜜蜂、Apis mellifera L.、α-葡萄糖苷酶 III 的纯化和底物特异性”Biosci，生物技术。生物化学.. 65. 1610-1616 (2001)

S. Hamada, K. Nozaki, H. Ito, Y, Yoshimoto, H. Yoshida, S. Hiraga, S. Onodera, M. Honma, Y. Takeda, H. Matsui: "Two starch branching enzymes isoforms occur in different fractions of developing seeds of kidney bean"Biochem. J.. 359. 23-34 (2001)

S. Hamada、K. Nozaki、H. Ito、Y、Yoshimoto、H. Yoshida、S. Hiraga、S. Onodera、M. Honma、Y. Takeda、H. Matsui：“两种淀粉分支酶亚型存在于不同的组分中

T. Mizuno, H. Mori, M. Nishino, H. Ito, H. Matsui, A. Kimura, M. Honma, S. Chiba: "Isolation and Sequence of a Putative a-Glueosidase Gene from Brevibacterium fuscum var. dextranlyticum strain 0407"J. Appl. Glycosci.. 48. 287-291 (2001)

T. Mizuno、H. Mori、M. Nishino、H. Ito、H. Matsui、A. Kimura、M. Honma、S. Chiba：“从短杆菌右旋糖酐变种菌株中推定的 α-葡萄糖苷酶基因的分离和序列

T.Mizuno ら: "Isolation and Sequence of a Putative α-Glurosiclase Gene from Brevibacterium fuscum var dextranixticum Strain 0467"Journal of Applied Glycoscience. 48. 287-291 (2001)

T. Mizuno 等人：“来自右旋短杆菌菌株 0467 的推定 α-葡萄糖苷酶基因的分离和序列”应用糖科学杂志 48. 287-291 (2001)。