Department of Food Science, Faculty of Sciences Copenhagen University, Rolighedsvej 30, 1958, Frederiksberg C, Denmark.
BMC Microbiol. 2012 Aug 13;12:174. doi: 10.1186/1471-2180-12-174.
During the transmission route from poultry to the human host, the major foodborne pathogen C. jejuni may experience many types of stresses, including low pH caused by different acids. However, not all strains are equally sensitive to the stresses. The aim of this study was to investigate the response to acid stress of three sequenced C. jejuni strains with different acid tolerances using HCl and acetic acid.
Two-dimensional gel electrophoresis was used for proteomic analysis and proteins were radioactively labelled with methionine to identify proteins only related to acid exposure. To allow added radioactive methionine to be incorporated into induced proteins, a modified chemically defined broth was developed with the minimal amount of methionine necessary for satisfactory growth of all strains. Protein spots were analyzed using image software and identification was done with MALDI-TOF-TOF. The most acid-sensitive isolate was C. jejuni 327, followed by NCTC 11168 and isolate 305 as the most tolerant. Overall, induction of five proteins was observed within the pI range investigated: 19 kDa periplasmic protein (p19), thioredoxin-disulfide (TrxB), a hypothetical protein Cj0706 (Cj0706), molybdenum cofactor biosynthesis protein (MogA), and bacterioferritin (Dps). Strain and acid type dependent differences in the level of response were observed. For strain NCTC 11168, the induced proteins and the regulator fur were analysed at the transcriptomic level using qRT-PCR. In this transcriptomic analysis, only up-regulation of trxB and p19 was observed.
A defined medium that supports the growth of a range of Campylobacter strains and suitable for proteomic analysis was developed. Mainly proteins normally involved in iron control and oxidative stress defence were induced during acid stress of C. jejuni. Both strain and acid type affected sensitivity and response.
在从禽类到人类宿主的传播途径中,主要食源性病原体空肠弯曲菌可能会经历多种类型的应激,包括不同酸引起的低 pH 值。然而,并非所有菌株对这些应激都同样敏感。本研究旨在使用 HCl 和乙酸研究三种不同耐酸能力的测序空肠弯曲菌菌株对酸应激的反应。
使用二维凝胶电泳进行蛋白质组学分析,并使用放射性标记的甲硫氨酸对仅与酸暴露相关的蛋白质进行标记。为了允许添加的放射性甲硫氨酸掺入诱导的蛋白质中,开发了一种改良的化学定义培养基,其中含有满足所有菌株生长所需的最小量的甲硫氨酸。使用图像软件分析蛋白质斑点,并使用 MALDI-TOF-TOF 进行鉴定。最敏感的分离株是空肠弯曲菌 327,其次是 NCTC 11168 和分离株 305,最耐受。总体而言,在研究的 pI 范围内观察到五种蛋白质的诱导:19 kDa 周质蛋白(p19)、硫氧还蛋白-二硫化物(TrxB)、一种假定蛋白 Cj0706(Cj0706)、钼辅因子生物合成蛋白(MogA)和菌铁蛋白(Dps)。观察到菌株和酸类型依赖性的反应水平差异。对于 NCTC 11168 菌株,使用 qRT-PCR 在转录组水平上分析了诱导蛋白和调控因子 fur。在这个转录组分析中,只观察到 trxB 和 p19 的上调。
开发了一种支持一系列弯曲菌菌株生长并适合蛋白质组学分析的定义培养基。空肠弯曲菌在酸应激下主要诱导参与铁控制和氧化应激防御的蛋白质。菌株和酸类型均影响敏感性和反应。