Salmonella Utilizes D-Glucosaminate via a Mannose Family Phosphotransferase System Permease and Associated Enzymes
Salmonella Utilizes D-Glucosaminate via a Mannose Family Phosphotransferase System Permease and Associated Enzymes
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DOI：
10.1128/jb.00290-13
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发表时间：
2013-09-01
影响因子：
3.2
通讯作者：
Hoover, Timothy R.
中科院分区：
文献类型：
--
作者：
Miller, Katherine A.;Phillips, Robert S.;Hoover, Timothy R.
肠道沙门氏菌是一种利用多种碳源的全球重要的细菌性食源性病原体。我们在这里报告说，沙门氏菌肠道亚种。鼠伤寒沙门氏菌（S. Typhimurium）通过先前未表征的甘露糖家族磷酸转移酶系统（PTS）通透酶使用D-氨基葡糖（2-氨基-2-脱氧-D-葡糖酸）作为碳源和氮源，并且我们将编码通透酶dgaABCD的基因命名为（D-氨基葡糖PTS通透酶组分EIIA、EIIB、EIIC和EIID）。dga操纵子中的另外两个基因（dgaE和dgaF）是野生型S生长所必需的。含D-氨基葡萄糖的鼠伤寒沙门氏菌。dgaABCDEF的转录依赖于RpoN（sigma（54））和我们命名为dgaR的RpoN依赖性激活基因。将lac启动子控制下的带有dgaABCDEF的质粒导入大肠杆菌菌株DH 5 α、BL 21和JM 101中，使这些菌株在含有D-氨基葡萄糖作为唯一碳源和氮源的基本培养基上生长。生化和遗传数据支持一种分解代谢途径，其中D-氨基葡萄糖在穿过细胞膜时被DgaABCD在C-6位置磷酸化。DgaE将所得D-氨基葡糖酸-6-磷酸转化为2-酮-3-脱氧葡糖酸6-磷酸（KDGP），其随后被醛缩酶DgaF裂解以形成甘油醛-3-磷酸和丙酮酸。DgaF催化与Entner-Doudoroff途径中的KDGP醛缩酶Eda催化的反应相同的反应，并且这两种酶可以在各自的途径中相互取代。综合微生物基因组数据库的检查显示，dga基因的直系同源物主要限于某些肠道细菌和厚壁菌门中的少数物种。
Salmonella enterica is a globally significant bacterial food-borne pathogen that utilizes a variety of carbon sources. We report here that Salmonella enterica subsp. enterica serovar Typhimurium (S. Typhimurium) uses D-glucosaminate (2-amino-2-deoxy-D-gluconic acid) as a carbon and nitrogen source via a previously uncharacterized mannose family phosphotransferase system (PTS) permease, and we designate the genes encoding the permease dgaABCD (D-glucosaminate PTS permease components EIIA, EIIB, EIIC, and EIID). Two other genes in the dga operon (dgaE and dgaF) were required for wild-type growth of S. Typhimurium with D-glucosaminate. Transcription of dgaABCDEF was dependent on RpoN (sigma(54)) and an RpoN-dependent activator gene we designate dgaR. Introduction of a plasmid bearing dgaABCDEF under the control of the lac promoter into Escherichia coli strains DH5 alpha, BL21, and JM101 allowed these strains to grow on minimal medium containing D-glucosaminate as the sole carbon and nitrogen source. Biochemical and genetic data support a catabolic pathway in which D-glucosaminate, as it is transported across the cell membrane, is phosphorylated at the C-6 position by DgaABCD. DgaE converts the resulting D-glucosaminate-6-phosphate to 2-keto-3-deoxygluconate 6-phosphate (KDGP), which is subsequently cleaved by the aldolase DgaF to form glyceraldehyde-3-phosphate and pyruvate. DgaF catalyzes the same reaction as that catalyzed by Eda, a KDGP aldolase in the Entner-Doudoroff pathway, and the two enzymes can substitute for each other in their respective pathways. Examination of the Integrated Microbial Genomes database revealed that orthologs of the dga genes are largely restricted to certain enteric bacteria and a few species in the phylum Firmicutes.