Figure 8. Zwf1 and the Ctt1 catalase activity is increased in Maf1-deficient mutant.

Yeast cells logarithmically growing in YPD (A, B, C) or YPGly (A) medium were harvested at D₆₀₀ ≈ 1.0. (A) For Zwf1 activity assay, NADH breakdown was measured at 340 nm in time at 30°C. For catalase activity (B), hydrogen peroxide decomposition in reaction mixtures containing yeast cell-free extracts was monitored as a change in absorbance at 240 nm in time at 30°C. Results are presented as total mean enzymatic activity from five independent biological replicates with standard deviation expressed as µmol·min⁻¹·mg⁻¹ protein. Asterisk (*) indicate P-value <0.05 according to Student's t-test for biological replicates. (C) Glutathione GSSG/GSH ratio in maf1Δ and rpc128-1007 does not indicate oxidative stress. GSSG, GSH and total glutathione levels were measured according to Quantification kit for oxidized and reduced glutathione (Sigma–Aldrich, 38185) from the kinetic method in agreement with the manufacturer's protocol. Yeast strains were grown in rich medium supplemented with 2% glucose until D₆₀₀ ≈ 1.0. The absorbance was obtained for four biological samples assayed in duplicate at 405 nm. [GSSG]/[GSH] ratio was measured separately for each sample and averaged. Results are shown as a mean value with standard deviation from four biological replicates. Asterisk (*) indicate P-value <0.05 according to Student's t-test for biological replicates.