Studies on ecological activity and enzymes of periodontopathogens

牙周病原菌生态活性及酶学研究

• 批准号: 63570883
• 负责人: NAKAMURA Takeshi
• 金额: $ 1.28万
• 依托单位: Matsumoto Dental College, Faculty of Dentistry
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570883/
• 关键词: Periodontopathogen; Antibacterial ability; Bacterial enzyme; Heparinase; Hyaluronidase; Neuraminidase; Protease; 歯周病原菌; 口腔Bacteroides; 口腔スピロヘ-タ; 細菌酵素; Neuraminidase; 生態適生物活性; ヘパリナーゼ; ヒアルロニダーゼ

1.Ecological effects, and purification and properties of antibacterial substance of Capnocytophaga ochracea. Cell-associated antibacterial substance of C. ochracea No.26 and M-12 was purified 83-fold. Molecular weight of this substance was 100,000 and its isoelectric point was 8.0. The substance inhibited bacteriostatically the growth of oral streptococci and Propionibacterium acnes.2.Purification and properties of mucopolysaccharidases and neuraminidase of the periodontopathogens. (i)Heparinase of Bacteroides heparinolyticus was purified 1,920-fold from cell extracts. Molecular weight of the enzyme was 64,000 and its isoelectric point was 9.5. Optimum pH for the activity was 7.0. only heparin and heparan sulfate were degraded by the enzyme. (ii)Extracellular hyaluronidase of Bacteroides oralis was purified 12,000- fold. Molecular weight of the enzyme was 75,000 and its isoelectric point was 7.6. Optimum pH for the activity was 5.5. Only hyaluronic acid was degraded by the enzyme. (iii)Cell-associated neuraminidase of Bacteroides gingivalis was purified 230-fold. Molecular weight of the enzyme was 55,000 and its isoelectric point was 4.5. Optimum PH for the activity was 5.8. Red blood cells treated with the purified enzyme were agglutinated by cells of Bacteroidesintermedius. Since neuraminidase activity was found in B. heparinolyticus and oral Treponema denticola, purification of each enzyme was undertaken.3.Biological properties and protease activity of oral Treponema denticola. Collagenolytic, trypsin-like, and chymotrypsin-like activity of T. denticola isolated from lesions of periodontitis were found using synthetic substrates. Each enzyme could be separated by ion-exchange chromatography, which suggests the proteolytic system of this oraganism, is rather diverse.

1.赭藻碳噬菌的生态效应及抗菌物质的纯化与性质。对C. ochracea No.26和M-12的细胞相关抗菌物质进行了83倍纯化。该物质的分子量为100,000，等电点为8.0。该物质对口腔链球菌和痤疮丙酸杆菌的生长具有抑菌作用。牙周病原粘多糖酶和神经氨酸酶的纯化及性质研究。(i)从细胞提取物中纯化出溶肝杆菌的肝素酶1,920倍。酶的分子量为64000，等电点为9.5。最适pH值为7.0。该酶仅能降解肝素和硫酸肝素。（二）对口腔拟杆菌胞外透明质酸酶进行12000倍纯化。酶的分子量为75000，等电点为7.6。最适pH为5.5。只有透明质酸被酶降解。（iii）对牙龈拟杆菌细胞相关神经氨酸酶进行230倍纯化。酶的分子量为55000，等电点为4.5。最适PH为5.8。经纯化酶处理的红细胞被中间拟杆菌细胞凝集。由于在溶肝螺旋体和口腔密螺旋体中发现了神经氨酸酶活性，因此对每种酶进行了纯化。口腔密螺旋体生物学特性及蛋白酶活性研究。利用合成底物发现了从牙周炎病变中分离出的牙牙霉具有胶原溶解、胰蛋白酶样和凝乳胰蛋白酶样活性。每种酶都可以通过离子交换色谱分离，这表明该生物的蛋白质水解系统相当多样化。

项目成果

Takeshi Nakamura et al.: "Purification and Properties of Oral Treponema denticola Protease." Oral Microbiol Immunol.

Takeshi Nakamura 等人：“口腔密螺旋体蛋白酶的纯化和特性。”

Takeshi Nakamura et al.: "Purification and Properties of Antibacterial Agent from Capnocytophaga ochracea." Infection and Immunity.

Takeshi Nakamura 等人：“来自赭二氧化碳噬菌体的抗菌剂的纯化和特性”。

柴田幸永 他: 松本歯学. 14. 58-65 (1988)

Yukinaga Shibata 等人：松本牙科。14. 58-65 (1988)

Takeshi Nakamura et al.: "Purification and Properties of Bacteroides heparinolyticus Heparinase(Heparin lyase,EC4、2、2、7)." Journal of Clinical Microbiology. 26. 1070-1071 (1988)

Takeshi Nakamura 等人：“溶肝拟杆菌肝素酶的纯化和特性（肝素裂解酶，EC4, 2, 2, 7）。临床微生物学杂志 26. 1070-1071 (1988)”

柴田幸永,他: "Bacteroides oralisのヒアルロニダ-ゼの精製とその性状" 松本歯学. 14. 58-65 (1988)

Yukinaga Shibata 等人：“从口腔拟杆菌中纯化透明质酸酶及其特性”Matsumoto Dental Science 14. 58-65 (1988)。