Development of new antibiotics targeting cell wall biosynthesis of oral pathogenic bacteria

针对口腔病原菌细胞壁生物合成的新型抗生素的开发

• 批准号: 12557186
• 负责人: YAMASHITA Yoshihisa
• 金额: $ 7.62万
• 依托单位: Kyushu University (2003)Nihon University (2000-2002)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557186/
• 关键词: Streptococcus mutans; Actinobacillus actinomycetemcomitans; cell wall polysaccharide; antibiotics; S.mutans; A.actinomycetemcomitans; 莢膜様多糖; ABCトランスポーター; 齲蝕細菌; 歯周病細菌; 薬剤耐性; 口腔病原性細菌; 抗生剤; Actinobacillus actinomycetemcomitans; Streptococcus mu

(The 2000 fiscal year)The genes which were related to the synthesis of the capsule polysaccharide of A.actinomycetemconutans(Aa) and cell wall polysaccharide of S.mutans was cloned. Homologous research for the determined sequence of the cloned genes revealed that some of them seemed to encode glycosyltransferases.(The 2001 fiscal year)The detailed function of each gene product was analyzed with the expressed gene products in Escherichia coli. The result indicated that the gene product of rgpA which is located in the uppermost stream of rgp gene cluster was glycosyltransferase which catalyzes the transfer of the first rhamnose residue. Furthermore, it became to be obvious that the serotype specific cell wall polysaccharide was important for the drug tolerance of S.mutans by comparing the drug tolerance for antibiotics of wild type strain Xc and its mutant strain Xc41.(The 2002 fiscal year)The genes which encode GDP-D-mannose 4,6-dehydratase and GDP-keto-6-deoxy-D-mannose reductase were … More cloned from the chromosome of the serotype a Aa SUNYaB75 strain. In addition, construction of the variant mutant strains Aa which lost serotype a,c, and d capsular polysaccharide antigens was succeeded by the insertional inactivation. When the tolerance of these variants for several antimicrobial drugs was examined, the sensitivity for bacitracin and vancomycin also slightly increased in any variants.(The 2003 fiscal year)The antibodies were raised against the rgpC and rgpD gene products of S.mutans expressed in and purified from E.coli. Though S.mutans was cultivated in the presence of the obtained antibodies, it was not possible to effectively inhibit the cell wall polysaccharide synthesis. When the antibody was taken in liposome and added to the culture medium, the interference tendency in the cell wall polysaccharide synthesis was slightly recognized. In case of Aa, antibodies against ABC transporters concerning the synthesis of the polysaccharide were prepared. Every antibodies showed slight blocked effect on capusular polysaccharide synthesis. Less

(The 2000财政年度）克隆了与放线菌荚膜多糖（Aa）和变形链球菌细胞壁多糖合成相关的基因。对克隆基因的同源性研究表明，其中一些基因可能编码糖基转移酶。(The 2001财政年度），每个基因产物的详细功能进行了分析与表达的基因产物在大肠杆菌。结果表明，位于rgp基因簇上游的rgpA基因产物为糖基转移酶，催化第一个鼠李糖残基的转移。此外，通过比较野生型菌株Xc及其突变株Xc 41对抗生素的耐药性，可以看出血清型特异性细胞壁多糖对变形链球菌耐药性的重要性。(The编码GDP-D-甘露糖4，6-脱氢酶和GDP-酮-6-脱氧-D-甘露糖还原酶的基因是 ...更多信息 从血清型a Aa SUNYaB 75菌株的染色体上克隆。此外，通过插入失活成功构建了缺失血清型a、c和d荚膜多糖抗原的变异突变株Aa。当检查这些变体对几种抗菌药物的耐受性时，在任何变体中对杆菌肽和万古霉素的敏感性也略有增加。(The 2003财政年度）针对在大肠杆菌中表达并从大肠杆菌中纯化的变形链球菌的rgpC和rgpD基因产物产生抗体。虽然在所获得的抗体存在下培养变形链球菌，但不可能有效地抑制细胞壁多糖合成。当抗体被置于脂质体中并加入到培养基中时，对细胞壁多糖合成的干扰倾向被轻微地识别。在Aa的情况下，制备针对与多糖合成有关的ABC转运蛋白的抗体。各抗体对荚膜多糖的合成均有一定的抑制作用。少

项目成果

Shibata, Y. et al.: "Analysis of loci required for determination of serotype antigenicity in Streptococcus mutans and its clinical utilization"Journal of Clinical Microbiology. 41. 4107-4112 (2003)

Shibata, Y. 等人：“分析变形链球菌血清型抗原性所需的基因座及其临床应用”临床微生物学杂志。

Suzuki, N., Nakano, Y., Yoshida, Y., et al.: "Guanosine diphosphate-4-keto-6-deoxy-D-mannose reductase in the pathway for the synthesis of GDP-6-deoxy-D-talose in Actinobacillus actinomycetemcomitans"Eur.J.Biochem.. 269. 5963-5971 (2002)

Suzuki, N.、Nakano, Y.、Yoshida, Y.等人：“GDP-6-脱氧-D-合成途径中的鸟苷二磷酸-4-酮-6-脱氧-D-甘露糖还原酶

Y.Nakano et al.: "Thymidine diphosphate-6-deoxy-L-lyxo-4-hexulose reductase synthesis-."Journal of Biological Chemistry. 275巻10号. 6806-6812 (2000)

Y. Nakano 等人：“胸苷二磷酸-6-脱氧-L-lyxo-4-己酮糖还原酶合成-”。《生物化学杂志》，第 275 卷，第 10 期。6806-6812 (2000)

Shibata, Y., Yamashita, Y., Ozaki, K., et al.: "Expression and characterization of streptococcal rgp genes required for rhamnan synthesis in Escherichia coli"Infect.Immun.. 70. 2891-2898 (2002)

Shibata, Y.、Yamashita, Y.、Ozaki, K. 等人：“大肠杆菌中鼠李聚糖合成所需的链球菌 rgp 基因的表达和表征”Infect.Immun.. 70. 2891-2898 (2002)

Ozaki, K., Shibata, Y., Yamashita, Y., et al.: "A novel mechanism for glucose side-chain formation in rhamnose-glucose polysaccharide synthesis"FEBS Letters. 532. 159-163 (2002)

Ozaki, K.、Shibata, Y.、Yamashita, Y. 等人：“鼠李糖-葡萄糖多糖合成中葡萄糖侧链形成的新机制”FEBS Letters。