STUDY ON METABOLIC SYSTEMS FOR PEPTIDES AND AMINO ACIDS OF PERIODONTAL PATHOGENS, PORPHYROMONAS AND PREVOTELLA.

牙周病原菌、卟啉单胞菌和普氏菌肽和氨基酸代谢系统的研究。

• 批准号: 11470386
• 负责人: TAKAHASHI Nobuhiro
• 金额: $ 6.66万
• 依托单位: Tohoku 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-11470386/
• 关键词: P. gingivalis; P. intermedia; PEPTIDE METABOLISM; ASPARTATE; GLUTAMATE; DIPEPTIDE; METABOLIC ENZYMES; PCR-RFLP; Porphyromonas gingivalis; Prevotella intermedia; Fusobacterium nucleatum; PCR-RELP法; Dipeptidyl peptidase; Dipeptidase; アミノ酸代謝

1. Peptides and amino acids as sources for the growth and the energy production of Porphyromonas gingivalis (Pg) and Prevotella intermedia(Pi)Periodontal pathogens, P. gingivalis and P. intermedia utilized peptides, especially peptides containing aspartate and/or glutamate, as the sources of their growth and energy production. In addition, P. gingivalis preferentially utilized aspartylaspartate and glutamylglutamate, while P. intermedia utilized aspartate and aspartylaspartate. The observation that both bacteria had no dipeptidase activity on their cell surfaces indicates that they incorporate dipeptides through a dipeptide-specific transport system. These findings suggest the existence of segregation or competition between P. gingivalis and P. intermedia for their nutrients.2. Metabolic pathways for peptides and amino acids of P. gingivalis and P. intermediaMetabolic system for peptides and amino acids of P. gingivalis and P. intermedia consists of the following steps: (l) peptide uptake (2) hydrolysis of peptide into amino acid (3) deamination of amino acid into 2-oxoacid (4) oxidation of 2-oxoacid into acyl-CoA and (5) production of short-chain fatty acid. 2-oxoacid oxidoreductases, which catalyze the reaction (4), utilized an artificial electron acceptor, methylviologen. This property is similar to that of enzymes found in archea.3. Molecular biological method for identification of periodontal pathogensFrom the ecological point of view as described 1, a molecular biological method for bacterial identification, was developed to know the distribution in dental plaque of periodontal pathogens, P. gingivalis and P. intermedia. A new method, PCR-RFLP based on bacterial 16S ribosomal RNA gene, was found to be not only simple and fast but also more reliable than the conventional method such as biological and morphological tests.

1.牙龈卟啉单胞菌(Pg)和中间普氏杆菌(PI)牙周病原菌的生长和产能以多肽和氨基酸为主要来源，牙龈卟啉单胞菌和中间普里沃特氏菌利用多肽，特别是含有天冬氨酸和/或谷氨酸的多肽作为其生长和产能的来源。此外，牙龈假单胞菌优先利用天冬氨酸和谷氨酸，而中间假单胞菌则优先利用天冬氨酸和天冬氨酸。观察到这两种细菌的细胞表面都没有二肽酶活性，这表明它们通过二肽特异的运输系统结合了二肽。这些发现提示牙龈假单胞菌和中间假单胞菌之间存在营养分离或竞争。牙龈假单胞菌和中间假单胞菌多肽和氨基酸的代谢途径牙龈假单胞菌和中间假单胞菌的多肽和氨基酸代谢系统包括以下步骤：(L)摄取多肽；(2)将多肽水解为氨基酸；(3)氨基酸脱氨为2-氧基酸；(4)将2-氧基酸氧化为酰辅酶A；(5)合成短链脂肪酸。催化该反应的2-氧代酸氧化还原酶(4)利用了一种人造电子受体甲基紫精。这一特性与在古迹中发现的酶相似。牙周病原菌的分子生物学鉴定方法从生态学的角度出发，建立了一种细菌鉴定的分子生物学方法，以了解牙周病原菌、牙周病原菌和中间牙周炎病原菌在牙菌斑中的分布情况。基于细菌16S核糖体RNA基因的聚合酶链式反应-限制性片段长度多态性方法不仅简单、快速，而且比常规的生物学和形态检测方法更可靠。

项目成果

Yamamoto Y., Sato Y., Takahashi-Abbe S., Takahashi N., Kizaki H.: "Characterization of the Streptococcus mutans pyruvate formate-lyase (PFL)-activating enzyme gene by complementary reconstitution of the in vitro PFL-reactivating system"Infect. Immun.. 68.

Yamamoto Y.、Sato Y.、Takahashi-Abbe S.、Takahashi N.、Kizaki H.：“通过体外 PFL 重新激活系统的互补重建来表征变形链球菌丙酮酸甲酸裂解酶 (PFL) 激活酶基因

JingPing Hu: "Copidis Rhizoma in hibits growth and proteolytic activity of oral bacteria"Oral Dis. 6. 297-302 (2000)

胡静平：“黄连抑制口腔细菌的生长和蛋白水解活性”Oral Dis。

Shoko Takahashi-Abbe: "Inhibitory effect of sorbitol on sugar metabolism of Streptococcus mutans in vitro and on acid production in dental plaque in vivo"Oral Microbiol Immunol. 16. 94-99 (2001)

Shoko Takahashi-Abbe：“山梨醇对体外变形链球菌糖代谢和体内牙菌斑酸产生的抑制作用”口腔微生物免疫学。

Kaoru Saito: "Effects of glucose on formation of cytotoxic end-products and proteolytic activity of Prevotella intermedia, Prevotella nigrescens and Porphyromonas gingivalis"J Periodontal Res. 36. 355-360 (2001)

Kaoru Saito：“葡萄糖对中间普雷沃菌、黑普雷沃菌和牙龈卟啉单胞菌的细胞毒性终产物形成和蛋白水解活性的影响”J 牙周研究。

Takahashi N., Yamada T.: "Effect of pH on the glucose and lactate metabolisms of Actinomyces naeslundii"Oral Microbiol. Immunol.. 14. 60-65 (1999)

Takahashi N.、Yamada T.：“pH 对内氏放线菌葡萄糖和乳酸代谢的影响”口腔微生物。