ORIGINAL PAPER Removal pattern of vinasse phenolics by Phlebia rufa, characterization of an induced laccase and inhibition kinetics modeling Joana M. C. Fernandes . Irene Fraga . Rui M. F. Bezerra . Albino A. Dias Received: 11 September 2020 / Accepted: 21 March 2021 / Published online: 27 March 2021 Ó The Author(s), under exclusive licence to Springer Nature B.V. 2021 Abstract Vinasse from the distillation of winemak- ing residues is a wastewater characterized by high levels of aromatic compounds. Batch cultures of Phlebia rufa showed a significant (p \ 0.05) correla- tion 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 com- pound. Vinasse-induced laccase showed elec- trophoretic 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 activa- tion energy of 42.8 kJ mol -1 . Catalytic efficiency of 2,2 0 -azinobis(3-ethylbenzthiazoline-6-sulfonic acid) oxidation is about two orders of magnitude higher than 2,6-DMP oxidation, being their K m values 36.2 ± 2.6 lM and 303.0 ± 44.7 lM, respectively and k cat 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. Inhibi- tion 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 Introduction Biosynthesis of secondary aromatic compounds by plants occurs in response to the interaction between a particular genotype and environmental effects, e.g., edaphoclimatic stress conditions. Secondary aromatic compounds are found in most plant tissues, and many of them have been identified in several fruits (Velioglu et al. 1998), including grape berries from Vitis vinifera Supplementary Information The online version contains supplementary material available at https://doi.org/10.1007/ s10532-021-09936-z. J. M. C. Fernandes  I. Fraga  R. M. F. Bezerra  A. A. Dias (&) CITAB - Centre for the Research and Technology of Agro-Environmental and Biological Sciences, UTAD – Universidade de Tra ´s-os-Montes e Alto Douro, 5000-801 Vila Real, Portugal e-mail: jdias@utad.pt I. Fraga  R. M. F. Bezerra  A. A. Dias Department of Biology and Environment, UTAD – Universidade de Tra ´s-os-Montes e Alto Douro, 5000-801 Vila Real, Portugal 123 Biodegradation (2021) 32:287–298 https://doi.org/10.1007/s10532-021-09936-z