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Studies on the basis of the glucosyltransfer activity of dextransucrase

基于葡聚糖蔗糖酶的糖基转移活性的研究

基本信息

批准号：
61560081
负责人：
KOBAYASHI Mikihiko
金额：
$ 1.22万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1986
资助国家：
日本
起止时间：
1986 至 1987
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-61560081/
关键词：
Dextransucrase Glucosyltransfer reaction Chemical modification Affinity labeling 多糖生合成

项目摘要

Mechanism of the dextransucrase reaction is important to clarify because of the understanding of the polysaccharide biosynthesis. In this research, we focused our attention on the modification of dextransucrase from Leuconostoc mesenteroides reffering to the change in the activity of glucosyltransfer. The following results were obtained; binding of the substrates, sucrose and dextran, occurred at the separate sites in the dextransucrase molecule for the acceptor and donor molecules. Dextransucrase was inactivated by the modification with water-soluble carbodiimide (EDC) and diethylphro-carbonate. Moreover, an affinity labeling of dextransucrase was conducted by orthophthalaldehyde and epoxide derivative of <alpha>-cyclodextrin (CD-epoxd). Effective inactivation of this enzyme with EDC and CD-epoxd suggested that a carboxyl group was important for the activity. A new function of dextransucrase could be provided by the extensive survey on the modification of respective acceptor- and/or donor-sites of the enzyme.

由于对多糖生物合成的理解，葡聚糖蔗糖酶反应的机理的澄清是重要的。本研究以肠膜明串珠菌的葡聚糖蔗糖酶为研究对象，对葡聚糖蔗糖酶进行修饰，改变葡糖基转移酶的活性。获得了以下结果;底物蔗糖和葡聚糖的结合发生在葡聚糖蔗糖酶分子中受体和供体分子的不同位点。通过水溶性碳二亚胺（EDC）和碳酸二乙酯修饰使右旋糖酐蔗糖酶失活。此外，葡聚糖蔗糖酶的亲和标记进行邻苯二甲醛和环氧衍生物的<alpha>-环糊精（CD-环氧）。用EDC和CD-环氧化合物有效灭活该酶表明羧基对活性是重要的。通过对葡聚糖蔗糖酶的受体和/或供体位点的修饰的广泛研究，可以提供葡聚糖蔗糖酶的新功能。