Metabolism of sugars by genetically diverse species of oral Leptotrichia.
Metabolism of sugars by genetically diverse species of oral Leptotrichia.
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DOI：
10.1111/j.2041-1014.2011.00627.x
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发表时间：
2012-02
影响因子：
3.7
通讯作者：
Pikis A
中科院分区：
文献类型：
--
作者：
Thompson J;Pikis A
颊纤毛菌ATCC 14201是一种存在于牙齿表面的口腔生物膜中的革兰氏阴性厌氧杆状细菌。该生物体的测序基因组揭示了三个连续的基因位点：Lebu_1525，1526和1527。这些基因的翻译产物分别与磷酸-α-葡萄糖苷酶（Pagl）、调节蛋白（GntR）和磷酸烯醇化糖转运蛋白（EIICB）具有显著的同源性。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，L. buccalis发酵各种各样的碳水化合物，包括蔗糖的五种异构体中的四种。在异构双糖上生长引发了大小与Lebu_1525编码的50 kDa多肽的表达。后者的基因被克隆，并表达的蛋白质纯化到同质从大肠杆菌TOP 10细胞。在NAD+和Mn ~（2+）两种辅因子存在下，该酶可水解对硝基苯基-α-吡喃葡萄糖苷6-磷酸（pNPα G6 P），后者是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，该酶可以归属于糖基水解酶超家族的家族4的磷酸-α-葡糖苷酶（Pagl）进化枝。我们认为Lebu_1527和1525的产物催化L.颊肌。最近描述了四种遗传多样性，但16 S rDNA相关的Leptotrichia种：L.古德费洛维湖hofstadii，L. shahii和L.瓦代。这些新物种的表型性状，就碳水化合物的利用，也已确定。
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.
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