Malone Aaron S, Chung Yoon-Kyung, Yousef Ahmed E
Department of Food Science and Technology, The Ohio State University, 2015 Fyffe Road, Columbus, OH 43210, USA.
Appl Environ Microbiol. 2006 Apr;72(4):2661-71. doi: 10.1128/AEM.72.4.2661-2671.2006.
Seventeen Escherichia coli O157:H7 strains were treated with ultrahigh pressure at 500 MPa and 23 +/- 2 degrees C for 1 min. This treatment inactivated 0.6 to 3.4 log CFU/ml, depending on the strain. The diversity of these strains was confirmed by pulsed-field gel electrophoresis (PFGE) analysis, and there was no apparent association between PFGE banding patterns and pressure resistance. The pressure-resistant strain E. coli O157:H7 EC-88 (0.6-log decrease) and the pressure-sensitive strain ATCC 35150 (3.4-log decrease) were treated with a sublethal pressure (100 MPa for 15 min at 23 +/- 2 degrees C) and subjected to DNA microarray analysis using an E. coli K-12 antisense gene chip. High pressure affected the transcription of many genes involved in a variety of intracellular mechanisms of EC-88, including the stress response, the thiol-disulfide redox system, Fe-S cluster assembly, and spontaneous mutation. Twenty-four E. coli isogenic pairs with mutations in the genes regulated by the pressure treatment were treated with lethal pressures at 400 MPa and 23 +/- 2 degrees C for 5 min. The barotolerance of the mutants relative to that of the wild-type strains helped to explain the results obtained by DNA microarray analysis. This study is the first report to demonstrate that the expression of Fe-S cluster assembly proteins and the fumarate nitrate reductase regulator decreases the resistance to pressure, while sigma factor (RpoE), lipoprotein (NlpI), thioredoxin (TrxA), thioredoxin reductase (TrxB), a trehalose synthesis protein (OtsA), and a DNA-binding protein (Dps) promote barotolerance.
将17株大肠杆菌O157:H7菌株在500兆帕、23±2摄氏度下进行超高压处理1分钟。根据菌株不同,该处理使细菌数量减少0.6至3.4个对数CFU/毫升。通过脉冲场凝胶电泳(PFGE)分析证实了这些菌株的多样性,并且PFGE条带模式与耐压性之间没有明显关联。将耐压菌株大肠杆菌O157:H7 EC - 88(减少0.6个对数)和压力敏感菌株ATCC 35150(减少3.4个对数)用亚致死压力(在23±2摄氏度下100兆帕处理15分钟)处理,然后使用大肠杆菌K - 12反义基因芯片进行DNA微阵列分析。高压影响了EC - 88多种细胞内机制中许多基因的转录,包括应激反应、硫醇 - 二硫键氧化还原系统、铁硫簇组装和自发突变。对24对在压力处理调控基因中发生突变的大肠杆菌同基因对在400兆帕、23±2摄氏度下进行致死压力处理5分钟。突变体相对于野生型菌株的耐压性有助于解释DNA微阵列分析获得的结果。本研究首次表明,铁硫簇组装蛋白和延胡索酸硝酸盐还原酶调节因子的表达会降低耐压性,而σ因子(RpoE)、脂蛋白(NlpI)、硫氧还蛋白(TrxA)、硫氧还蛋白还原酶(TrxB)、海藻糖合成蛋白(OtsA)和一种DNA结合蛋白(Dps)则促进耐压性。