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耐铝荧光假单胞菌:活性氧毒性与增强的烟酰胺腺嘌呤二核苷酸磷酸(NADPH)生成

Aluminum-tolerant Pseudomonas fluorescens: ROS toxicity and enhanced NADPH production.

作者信息

Singh Ranji, Beriault Robin, Middaugh Jeffrey, Hamel Robert, Chenier Daniel, Appanna Vasu D, Kalyuzhnyi Sergey

机构信息

Department of Chemistry and Biochemistry, Laurentian University, Sudbury, ON P3E2C6, Canada.

出版信息

Extremophiles. 2005 Oct;9(5):367-73. doi: 10.1007/s00792-005-0450-7. Epub 2005 Jun 22.

Abstract

Aluminum (Al) triggered a marked increase in reactive oxygen species (ROS) such as O(2) (-) and H(2)O(2) in Pseudomonas fluorescens. Although the Al-stressed cells were characterized with higher amounts of oxidized lipids and proteins than controls, NADPH production was markedly increased in these cells. Blue native polyacrylamide gel electrophoresis (BN-PAGE) analyses coupled with activity and Coomassie staining revealed that NADP(+) -dependent isocitrate dehydrogenase (ICDH, E.C. 1.1.1.42) and glucose-6-phosphate dehydrogenase (G6PDH, E.C. 1.1.1.49) played a pivotal role in diminishing the oxidative environment promoted by Al. These enzymes were overexpressed in the Al-tolerant microbes and were modulated by the presence of either Al or hydrogen peroxide (H(2)O(2)) or menadione. The activity of superoxide dismutase (SOD, E.C. 1.15.1.1), an enzyme known to combat ROS stress was also increased in the cells cultured in millimolar amounts of Al. Hence, Al-tolerant P. fluorescens invokes an anti-oxidative defense strategy in order to survive.

摘要

铝(Al)引发荧光假单胞菌中活性氧(ROS)如超氧阴离子(O₂⁻)和过氧化氢(H₂O₂)显著增加。尽管与对照相比,铝胁迫细胞中氧化脂质和蛋白质的含量更高,但这些细胞中烟酰胺腺嘌呤二核苷酸磷酸(NADPH)的产量显著增加。蓝色非变性聚丙烯酰胺凝胶电泳(BN-PAGE)分析结合活性和考马斯亮蓝染色表明,依赖烟酰胺腺嘌呤二核苷酸磷酸(NADP⁺)的异柠檬酸脱氢酶(ICDH,E.C. 1.1.1.42)和葡萄糖-6-磷酸脱氢酶(G6PDH,E.C. 1.1.1.49)在减轻铝促进的氧化环境中起关键作用。这些酶在耐铝微生物中过表达,并受铝、过氧化氢(H₂O₂)或甲萘醌的存在调节。超氧化物歧化酶(SOD,E.C. 1.15.1.1)是一种已知可对抗ROS应激的酶,在以毫摩尔量的铝培养的细胞中其活性也增加。因此,耐铝荧光假单胞菌为了生存会调用一种抗氧化防御策略。

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