Regulation Of Sugar Transport And Metabolism In Oral Bacteria

口腔细菌中糖运输和代谢的调节

• 批准号: 8344108
• 负责人: john thompson
• 金额: $ 34.44万
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8344108
• 关键词: Address; Amino Acids; Anaerobic Bacteria; Bacteria; Benign; Biochemistry; Butyric Acids; Carbohydrates; Carrier Proteins; Cells; Dental caries; Disaccharides; Energy-Generating Resources; Enzymes; Escherichia coli; Etiology; Exhibits; Family; Fermentation; Fusobacteria; Genes; Genetic; Gingivitis; Growth; Hydrolase; Hydrolysis; Investigation; Ions; Isomerism; Journals; Lactic acid; Lactobacillus casei; Leptothrix; Leptotrichia; Metabolic; Metabolism; Microbial Biofilms; Mouth Diseases; Operon; Oral; Organism; Pathogenicity; Pathway interactions; Periodontitis; Property; Propionic Acids; Proteins; Publications; Pyruvate; Recombinant DNA; Regulation; Sequence Alignment; Shapes; Streptococcus; Sucrose; Sulfhydryl Compounds; Sulfides; Translations; analog; cofactor; comparative; cytotoxic; genetic regulatory protein; genome sequencing; glucosidase; gram negative anaerobic rods; inorganic phosphate; microbial; oral bacteria; oral biofilm; oral biology; oral microbiome; pathogen; polypeptide; programs; sugar; tooth surface; trait

Leptotrichia buccalis ATCC 14201 is a Gram-negative, anaerobic rod-shaped bacterium resident in oral biofilm at the tooth surface. The sequenced genome of this organism reveals three contiguous genes at loci: Lebu_1525,1526 and 1527. The translation products of these genes exhibit significant homology with phospho-α-glucosidase (Pagl), a regulatory protein (GntR) and a phosphoenol pyruvate-dependent sugar transport protein (EIICB), respectively. In non-oral bacterial species, these genes comprise the sim operon that facilitates sucrose isomer metabolism. Growth studies showed that L. buccalis fermented a wide variety of carbohydrates, including four of the five isomers of sucrose. Growth on the isomeric disaccharides elicited expression of a 50kDa polypeptide comparable in size to that encoded by Lebu_1525. The latter gene was cloned, and the expressed protein was purified to homogeneity from Escherichia coli TOP 10 cells. In the presence of two cofactors, NAD+ and Mn2+ ion, the enzyme readily hydrolyzed p-nitrophenyl-α-glucopyranoside 6-phosphate (pNPαG6P), a chromogenic analog of the phosphorylated isomers of sucrose. By comparative sequence alignment, immuno-reactivity and signature motifs, the enzyme can be assigned to the phospho-α-glucosidase (Pagl) clade of Family 4 of the glycosyl hydrolase super family. We suggest that the products of Lebu_1527 and 1525, catalyze the phosphorylative translocation and hydrolysis of sucrose isomers in L. buccalis, respectively. Four genetically diverse, but 16S rDNA related species of Leptotrichia have recently been described: L. goodfellowii, L. hofstadii, L. shahii and L. wadei. The phenotypic traits of these new species, with respect to carbohydrate utilization, have also been determined.The results of our investigation have been submitted, and accepted, for publication in the journal MOLECULAR ORAL MICROBIOLOGY.

颊细毛菌 ATCC 14201 是一种革兰氏阴性厌氧杆状细菌，存在于牙齿表面的口腔生物膜中。该生物体的基因组测序揭示了位点上的三个连续基因：Lebu_1525、1526和1527。这些基因的翻译产物与磷酸-α-葡萄糖苷酶（Pagl）、调节蛋白（GntR）和磷酸烯醇丙酮酸依赖性蛋白表现出显着的同源性。糖转运蛋白（EIICB）。在非口腔细菌物种中，这些基因包含促进蔗糖异构体代谢的 sim 操纵子。生长研究表明，颊乳杆菌发酵多种碳水化合物，包括蔗糖五种异构体中的四种。在异构二糖上的生长引发了 50kDa 多肽的表达，其大小与 Lebu_1525 编码的多肽相当。后一个基因被克隆，表达的蛋白质从大肠杆菌TOP 10细胞中纯化至同质。在两种辅因子 NAD+ 和 Mn2+ 离子存在的情况下，该酶很容易水解对硝基苯基-α-吡喃葡萄糖苷 6-磷酸 (pNPαG6P)，一种蔗糖磷酸化异构体的显色类似物。通过比较序列比对、免疫反应性和特征基序，该酶可归属于糖基水解酶超家族第 4 族的磷酸-α-葡萄糖苷酶 (Pagl) 进化枝。我们认为Lebu_1527和1525的产物分别催化L. buccalis中蔗糖异构体的磷酸化易位和水解。最近描述了四种遗传多样性但 16S rDNA 相关的纤毛菌属物种：L. goodfellowii、L. hofstadii、L. shahii 和 L. wadei。这些新物种在碳水化合物利用方面的表型特征也已确定。我们的研究结果已提交并被接受，将在《分子口腔微生物学》杂志上发表。