Abstract
Commensal and pathogenic fungi are exposed to hydrogen peroxide (H2O2) produced by macrophages of the host. Pathogenic fungi counteract the harmful effects of H2O2 with the enzyme catalase (EC 1.11.1.6), which decomposes two molecules of H2O2 to two molecules of H2O and O2. Contribution of antioxidant systems on fungal virulence is actively studied. Measurement of catalase activity can contribute to the elucidation of the factors that influence the regulation of this pivotal enzyme. Here we describe a simple spectrophotometric method in which the activity of catalase is measured in total yeast extracts. Decomposition of H2O2 by the yeast extract is followed by the decrease in absorbance at 240 nm. The difference in absorbance through time (ΔA240) is inferred as the measure of catalase activity.
Keywords: Catalase, Candida, Glabrata, Yeast, CTA1
Materials and Reagents
Equipment
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Recipes
Acknowledgments
This protocol is based on the methodology reported by Aebi (1984), and by Weydert and Cullen (2010). Our adapted method was first published in Cuellar-Cruz et al. (2009). This work was funded by a CONACYT grant no. CB-2010-153929 to A.D.L.P. Finally, we thank Guadalupe Gutierrez-Escobedo for technical assistance.
References
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