Zentralinstitut für Ernährungs- und Lebensmittelforschung (ZIEL), Abteilung Mikrobiologie, Technische Universität München, Freising, Germany.
Appl Environ Microbiol. 2011 Jan;77(1):281-90. doi: 10.1128/AEM.01403-10. Epub 2010 Oct 29.
Only three pathogenic bacterial species, Salmonella enterica, Clostridium perfringens, and Listeria monocytogenes, are able to utilize both ethanolamine and 1,2-propanediol as a sole carbon source. Degradation of these substrates, abundant in food and the gut, depends on cobalamin, which is synthesized de novo only under anaerobic conditions. Although the eut, pdu, and cob-cbi gene clusters comprise 40 kb, the conditions under which they confer a selection advantage on these food-borne pathogens remain largely unknown. Here we used the luciferase reporter system to determine the response of the Salmonella enterica serovar Typhimurium promoters P(eutS), P(pocR), P(pduF), and P(pduA) to a set of carbon sources, to egg yolk, to whole milk, and to milk protein or fat fractions. Depending on the supplements, specific inductions up to 3 orders of magnitude were observed for P(eutS) and P(pduA), which drive the expression of most eut and pdu genes. To correlate these significant expression data with growth properties, nonpolar deletions of pocR, regulating the pdu and cob-cbi genes, and of eutR, involved in eut gene activation, were constructed in S. Typhimurium strain 14028. During exponential growth of the mutants 14028ΔpocR and 14028ΔeutR, 2- to 3-fold-reduced proliferation in milk and egg yolk was observed. Using the Caenorhabditis elegans infection model, we could also demonstrate that the proliferation of S. Typhimurium in the nematode is supported by an active ethanolamine degradation pathway. Taking these findings together, this study quantifies the differential expression of eut and pdu genes under distinct conditions and provides experimental evidence that the ethanolamine utilization pathway allows salmonellae to occupy specific metabolic niches within food environments and within their host organisms.
仅有三种致病菌,即沙门氏菌、产气荚膜梭菌和单核细胞增生李斯特菌,能够将乙醇胺和 1,2-丙二醇用作唯一的碳源。这些在食物和肠道中大量存在的底物的降解依赖于钴胺素,而钴胺素只能在厌氧条件下从头合成。尽管 eut、pdu 和 cob-cbi 基因簇包含 40kb,但它们使这些食源性病原体具有选择优势的条件在很大程度上仍然未知。在这里,我们使用荧光素酶报告系统来确定沙门氏菌血清型 Typhimurium 的 P(eutS)、P(pocR)、P(pduF)和 P(pduA)启动子对一组碳源、蛋黄、全脂奶以及乳蛋白或脂肪部分的响应。根据补充剂的不同,P(eutS)和 P(pduA)的表达可高达 3 个数量级,这驱动了大多数 eut 和 pdu 基因的表达。为了将这些显著的表达数据与生长特性相关联,我们构建了沙门氏菌 14028 株中 pocR(调节 pdu 和 cob-cbi 基因)和 eutR(参与 eut 基因激活)的非极性缺失突变体。在突变体 14028ΔpocR 和 14028ΔeutR 的指数生长期,观察到在牛奶和蛋黄中的增殖减少了 2-3 倍。使用秀丽隐杆线虫感染模型,我们还证明了乙醇胺降解途径的活性支持了沙门氏菌在线虫中的增殖。综合这些发现,本研究定量了 eut 和 pdu 基因在不同条件下的差异表达,并提供了实验证据,表明乙醇胺利用途径使沙门氏菌能够在食物环境和宿主生物体内占据特定的代谢生态位。