Bearson Bradley L, Lee In Soo, Casey Thomas A
Swine Odor and Manure Management Research Unit, USDA, ARS, National Soil Tilth Laboratory, NSRIC-2103, Ames, IA 50011, USA.
Department of Biotechnology, Hannam University, Daejeon 305-811, Republic of Korea.
Microbiology (Reading). 2009 Mar;155(Pt 3):805-812. doi: 10.1099/mic.0.022905-0.
Micro-organisms may simultaneously encounter multiple stresses in their environment. To investigate the protection that several known Escherichia coli O157 : H7 acid-resistance systems might provide against both oxidative and acid stress, the addition of diamide, a membrane-permeable thiol-specific oxidizing agent, or hydrogen peroxide were used concurrent with acid challenge at pH 2.5 to determine bacterial survival. The addition of either diamide or hydrogen peroxide decreased bacterial survival in a dose-dependent manner for E. coli O157 : H7 during challenge at pH 2.5 following overnight growth in LB MES pH 5.5 (acid-resistance system 1, AR1). In contrast, the presence of either glutamate or arginine during challenge provided significant protection against diamide- and hydrogen peroxide-induced oxidative stress during pH 2.5 acid challenge. Oxidative stress protection during acid challenge required gadC and adiA for the glutamate- (AR2) and arginine- (AR3) dependent acid-resistance systems, respectively. In addition, maximal protection against oxidative stress in the presence of glutamate required a low external pH (pH 2.5), since pH 5.5 did not protect. This study demonstrates that the glutamate- and arginine-dependent acid-resistance systems of E. coli O157 : H7 can simultaneously protect against oxidative stress during extreme acid challenge.
微生物在其环境中可能同时遭遇多种压力。为了研究几种已知的大肠杆菌O157 : H7耐酸系统对氧化应激和酸应激可能提供的保护作用,使用膜通透性硫醇特异性氧化剂二酰胺或过氧化氢,并在pH 2.5的酸应激挑战时同时添加,以确定细菌的存活率。在LB MES pH 5.5(耐酸系统1,AR1)中过夜培养后,在pH 2.5的挑战过程中,添加二酰胺或过氧化氢均以剂量依赖的方式降低了大肠杆菌O157 : H7的存活率。相比之下,在挑战过程中添加谷氨酸或精氨酸可在pH 2.5的酸应激挑战期间提供显著保护,抵御二酰胺和过氧化氢诱导的氧化应激。对于依赖谷氨酸的(AR2)和依赖精氨酸的(AR3)耐酸系统,酸应激挑战期间的氧化应激保护分别需要gadC和adiA。此外,在谷氨酸存在下对氧化应激的最大保护需要低外部pH(pH 2.5),因为pH 5.5没有保护作用。这项研究表明,大肠杆菌O157 : H7依赖谷氨酸和精氨酸的耐酸系统在极端酸应激挑战期间可同时抵御氧化应激。