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Laccases are copper-containing enzymes involved in the degradation of lignocellulosic materials and used in the treatment of phenol-containing wastewater. In this study we investigated the effect of culture conditions, i.e. submerged or semi-solid, and copper supplementation on laccase production by Trametes pubescens grown on coffee husk, soybean pod husk, or cedar sawdust. The highest specific laccase activity was achieved when the culture was conducted under submerged conditions supplemented with copper (5 mM), and using coffee husk as substrate. The crude extracts presented two laccase isoforms with molecular mass of 120 (Lac1) and 60 kDa (Lac2). Regardless of the substrate, enzymatic crude extract and purified fractions behaved similarly at different temperatures and pHs, most of them presented the maximum activity at 55 °C and a pH range between 2 and 3. In addition, they showed similar stability and electro-chemical properties. At optimal culture conditions laccase activity was 7.69±0.28 U mg-1 of protein for the crude extract, and 0.08±0.001 and 2.86±0.05 U mg-1 of protein for Lac1 and Lac2, respectively. In summary, these results show the potential of coffee husk as an important and economical growth medium to produce laccase, offering a new alternative use for this common agro-industrial byproduct.

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Anaerobic membrane bioreactors (AnMBRs) have demonstrated an excellent capability to treat domestic wastewater. However, biofouling reduces membrane permeability, increasing operational costs and overall energy demand. Soluble microbial products (SMPs) that build up on the membrane surface play a significant role in the biofouling. In this study, the production of SMPs in a 32 L submerged AnMBR operated at three different organic loads (3.0, 4.1 and 1.2 kg chemical oxygen demand (COD)/m3d for phases 1, 2 and 3, respectively) during long-term operation of the reactor (144, 83 and 94 days) were evaluated. The samples were taken from both the permeate and the sludge at three different heights (0.14, 0.44 and 0.75 m). Higher production of SMPs was obtained in phase 2, which was proportional to the membrane fouling. There were no statistically significant differences (p > 0.05) in the SMPs extracted from sludge at different heights among the three phases. In the permeate of phases 1, 2 and 3, the membrane allowed the removal of 56%, 70% and 64% of the SMP concentration in the sludge. SMPs were characterized by molecular weight (MW). A bimodal behavior was obtained, where fractions prevailed with an MW < 1 kDa, associated with SMPs as utilization-associated products (UAPs) caused fouling by the pore-blocking mechanism. The chemical analysis found that, in the SMPs, the unknown COD predominated over the known COD, such as carbohydrates and proteins. These results suggest that further studies in SMP characterization should focus on the unknown COD fraction to understand the membrane fouling in AnMBR systems better.