STRUCTURE OF ACTIVE SITE REGIONS ON EXO-GLYCOSIDASES

外切糖苷酶活性位点区域的结构

• 批准号: 07660087
• 负责人: MATSUI Hirokazu
• 金额: $ 1.34万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07660087/
• 关键词: alpha-glucosidase; dextranase; exo-type; Active site; cDNA; Primary structure; グルコシダーゼ; 化学修飾; イソマルトトリオース

Sugar beet alpha-glucosidase was modified with CBE.An analysis of an CBE-labeled peptide isolated from the digest of modified enzyme showed that the sequence around the essential amino acid was -DGIWIDMNE-. A cDNA encoding sugar beet alpha-glucosidase was cloned from a library constructed from mRNA of suspension cultured cells. The cDNA had an open reading frame encoding a polypeptide of 913 amino acid residues. The deduced amno acid sequence indicated relatively high homology in the range of 28.2 to 54.3% to those for other alpha-glucosidases.The gene encoding isomaltotrio-dextranase was cloned from Brevibacterium fuscum, and analyzed. The deduced amno acid sequence (641 residues) had 80% of homology to that of Arthrobacter dextranase.

用CBE对甜菜α-葡萄糖苷酶进行了修饰，对修饰后的酶消化产物中的CBE标记肽段进行了分析，发现其必需氨基酸的序列为-DGIWIDMNE-。从由悬浮培养细胞的mRNA构建的文库中克隆编码甜菜α-葡糖苷酶的cDNA。该cDNA具有一个开放的阅读框架，编码913个氨基酸残基的多肽。推导的氨基酸序列与其它α-葡聚糖酶的氨基酸序列同源性较高，在28.2 ~ 54.3%之间。推导的氨基酸序列（641个氨基酸残基）与节杆菌葡聚糖酶的氨基酸序列同源性为80%。

项目成果

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H.Matsui: "Cloning and Sequencing of a cDNA Encoding alpha-Glucosidase from Sugar Beet" Biosci.Biotech.Biochem.61, (in press). (1997)

H.Matsui：“编码甜菜 α-葡萄糖苷酶的 cDNA 的克隆和测序”Biosci.Biotech.Biochem.61，（印刷中）。