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Aluminum-tolerant Pseudomonas fluorescens: ROS toxicity and enhanced NADPH production
Authors:Ranji Singh  Robin Beriault  Jeffrey Middaugh  Robert Hamel  Daniel Chenier  Vasu D Appanna  Sergey Kalyuzhnyi
Institution:(1) Department of Chemistry and Biochemistry, Laurentian University, Sudbury, ON P3E2C6, Canada;(2) Northern Centre of Biotechnology and Clinical Research, Sudbury, ON P3E6B4, Canada;(3) Department of Chemical Enzymology, Moscow State University, Moscow, Russia
Abstract:Aluminum (Al) triggered a marked increase in reactive oxygen species (ROS) such as O2 and H2O2 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 (H2O2) 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.
Keywords:Al toxicity  ROS  Isocitrate dehydrogenase  Glucose 6 phosphate dehydrogenase  Superoxide dismutase  NADPH
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