Oxidative stress sensitivity and superoxide dismutase of a wild-type parent strain and a respiratory mutant of Candida albicans

Abstract

It is important to know responses of the pathogenic fungi to reactive oxygen species by which hosts protect themselves against fungal infection. In the present study, sensitivities to the superoxide radical (O2−) and superoxide dismutase (SOD) were compared between a wild‐type parent strain and a respiration‐deficient mutant of Candida albicans. When their survival was examined on an agar medium containing an intracellular O2− generator, paraquat (PQ), the parent strain was selectively killed by increasing the PQ concentration. In contrast, when cells of both strains were illuminated in a riboflavin solution, they exhibited similar sensitivity to O2− generated extracellularly by photo‐reduced riboflavin. There were no large differences in sensitivity to hydrogen peroxide in the two strains. Thus, the high tolerance of the mutant to PQ was suggested to result from low intracellular O2− generation by PQ due to the respiratory deficiency. It is generally accepted that fungal cells contain manganese (Mn)‐SOD in the mitochondria and copper and zinc (CuZn)‐SOD in the cytoplasm. Cyanide‐insensitive SOD activity (attributable to Mn‐SOD) was dominant in the parent strain throughout growth phases, whereas cyanide‐sensitive activity (attributable to CuZn‐SOD) occurred in the mutant. The activity bands of Mn‐ and CuZn‐SODs were clearly separated by electrophoresis of the cell extracts of both strains on non‐denaturing polyacrylamide gels. The electrophoretic profiles obtained were consistent with the results of the activity assay. These results showed that the respiratory deficiency affected oxidative stress sensitivity and SOD in C. albicans.

Functional analysis of the superoxide dismutase family in Aspergillus fumigatus

“…For the phenotypic analysis, we tested the sensitivity of the Δ sod mutants to oxidative stress conditions induced by menadione (up to 80 µM), paraquat (up to 8 mM). The sensitivity to extracellular superoxide ions of SOD mutants was analysed using the method described by Ito‐kuwa et al. (1999) with the addition of up to 1 mg ml −1 riboflavin to the medium in the presence of light.…”

“…For the phenotypic analysis, we tested the sensitivity of the Dsod mutants to oxidative stress conditions induced by menadione (up to 80 mM), paraquat (up to 8 mM). The sensitivity to extracellular superoxide ions of SOD mutants was analysed using the method described by Ito-kuwa et al (1999) with the addition of up to 1 mg ml -1 riboflavin to the medium in the presence of light. A positive control was performed using an YPD agar plate containing 2.45 ¥ 10 -3 M NBT chloride (Sigma) and 0.25 mg ml -1 riboflavin in order to confirm the production of superoxide radicals in the medium.…”

Antioxidant enzyme influences germination, stress tolerance, and virulence of Isaria fumosorosea

“…Cells of entomopathogenic fungi contain multiple cytosolic ROS detoxifying enzymes, such as superoxide dismutases, catalases, cytochrome c peroxidases, glutathione peroxidases, glutaredoxins and peroxiredoxins . SODs localized in the mitochondrial intermembrane space (CuZn SOD) and in the matrix (Mn SOD) form the first line of antioxidant defense against . Catalases are also one of the most important enzymatic systems used to degrade H 2 O 2 into water and oxygen, thereby lowering intracellular hydrogen peroxide levels .…”