Vinasse from the distillation of winemaking residues is a wastewater characterized by high levels of aromatic compounds. Batch cultures of Phlebia rufa showed a significant (p < 0.05) correlation between laccase activity and initial vinasse concentration. The pattern of biodegradation of hydroxybenzoic acids, hydroxycinnamic acids and flavonoids, assessed by HPLC-DAD, revealed that p-hydroxybenzoic acid is the most recalcitrant compound. Vinasse-induced laccase showed electrophoretic homogeneity and molecular weight of 62 kDa after being purified 21-fold. Optimum pH for oxidation of 2,6-dimethoxyphenol (2,6-DMP) was 3.5 and optimum temperature was 50 °C, with an activation energy of 42.8 kJ mol-1. Catalytic efficiency of 2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) oxidation is about two orders of magnitude higher than 2,6-DMP oxidation, being their Km values 36.2 ± 2.6 μM and 303.0 ± 44.7 μM, respectively and kcat values 486.1 s-1 and 179.6 s-1, respectively. Akaike information criterion and Akaike weights were used to discriminate inhibition models that best fitted 2,6-DMP oxidation in the presence of inhibitors. Inhibition constants of mixed-type inhibitors azide and fluoride, and competitive-type inhibitor chloride, showed the following inhibitors potency: azide > fluoride > chloride. Taken together, this study is consistent with the assumption that P. rufa could be a useful tool for aerobic degradation of phenolic-rich wastewaters.
Keywords: Aromatic compounds; Enzyme inhibition; Laccase kinetics; Phlebia rufa; Winemaking residues.