Regulation Of Sugar Transport And Metabolism In Oral Bacteria

口腔细菌中糖运输和代谢的调节

基本信息

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
项目类别：
财政年份：
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

项目摘要

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.

Buccalis ATCC 14201是一种革兰氏阴性，厌氧棒形细菌，驻留在牙齿表面的口腔生物膜中。该生物的测序基因组在基因座上揭示了三个连续基因：LEBU_1525,1526和1527。这些基因的翻译产物与磷酸化磷酸-α-葡萄糖苷酶（PAGL），调节蛋白（GNTR）和磷酸烯醇pyruvate糖蛋白（EII依赖型依赖型磷酸蛋白）表现出显着的同源性。在非口腔细菌物种中，这些基因包括促进蔗糖异构代谢的SIM操纵子。生长研究表明，buccalis发酵了多种碳水化合物，包括蔗糖的五个异构体中的四个。异构体二糖的生长引起的50KDA多肽的表达与LEBU_1525所编码的大小相当。将后一种基因克隆，并将表达的蛋白质纯化为大肠杆菌前10个细胞的同质性。在两个辅助因子的存在下，NAD+和MN2+离子，酶很容易水解的P-硝基苯基-α-葡萄吡烷糖苷6-磷酸（PNPαG6P），这是一种磷酸化异构体的成色性类似物。根据比较序列比对，免疫反应性和特征基序，可以将酶分配给糖基水解酶超级家族的家族4家族的磷酸化α-葡萄糖苷酶（PAGL）进化枝。我们建议分别催化Buccalis L. buccalis的蔗糖异构体的磷酸化易位和水解的产物。最近已经描述了四种遗传多样的，但瘦素的16S相关物种：L。Goodfelledii，L。Hofstadii，L。Shahii和L. Wadei。还确定了这些新物种的表型特征，也已经确定。我们的研究结果已被提交并接受，以在《分子口腔微生物学》杂志上发表。