Thermosensitive polymer-assisted extraction and purification of fungal laccase from citrus pulp wash effluent.

BACKGROUND: This study explores the use of liquid-liquid extraction with thermosensitive polymers for producing laccase (Lac) from Pleurotus sajor-caju. This process leverages liquid waste from the citrus industry, specifically pulp wash. The research delves into extractive fermentation and thermoseparation, both processes being facilitated by a polymer exhibiting a lower critical solution temperature transition. RESULTS: Key factors considered include the choice of polymer, its concentration, pH, separation temperature, and the behavior of the polymer-rich phase post-extractive fermentation concerning the lower critical solution temperature. Notably, under conditions of 45% by weight of Pluronic L-61 and pH 5.0 at 25 °C, the Lac resulted in an enhancement in the purification factor of 28.4-fold, compared with the Lac obtained directly from the fermentation process on the eighth day. There was an 83.6% recovery of the Lac enzyme in the bottom phase of the system. Additionally, the unique properties of Pluronic L-61, which can induce phase separation and also allow for thermoseparation, led to a secondary fraction (aqueous solution) of Lac with purification factor of 2.1 ± 0.1-fold (at 32 ± 0.9 °C and 30 ± 0.3 min without stirring) from the polymeric phase (top phase). Fourier-transform infrared analysis validated the separation data, particularly highlighting the α-helix content in the amide I region (1600-1700 cm-1 ). CONCLUSION: In summary, the insights from this study pave the way for broader industrial applications of these techniques, underscoring benefits like streamlined process integration, heightened selectivity, and superior separation efficacy. © 2023 Society of Chemical Industry.

Toward efficient electrocatalytic degradation of iohexol using active anodes: A laser-made versus commercial anodes.

The X-ray iodinated contrast medium iohexol is frequently detected in aquatic environments due to its high persistence and the inefficiency of its degradation by conventional wastewater treatments. Hence, the challenge faced in this study is the development of an alternative electrochemical treatment using active anodes. We investigate the oxidation of iohexol (16.42 mg L-1) using different operating conditions, focusing on the role of different mixed metal oxide anodes in the treatment efficiency. The electrocatalytic efficiency of the Ti/RuO2-TiO2 anode prepared using a CO2 laser heating and an ionic liquid is compared with Ti/RuO2-TiO2-IrO2 and Ti/IrO2-Ta2O5 commercial anodes. The hypochlorite ions generated by the anodes are also analyzed. The effect of the electrolyte composition (NaCl, Na2SO4, and NaClO4) and current density (15, 30, and 50 mA cm-2) on the iohexol degradation is also studied. The Ti/RuO2-TiO2 laser-made anode is more efficient than the commercial anodes. After optimizing experimental parameters, this anode removes 95.5% of iohexol in 60 min and displays the highest kinetic rate (0.059 min-1) with the lowest energy consumption per order (0.21 kWh m-3order-1), using NaCl solution as the electrolyte and applying 15 mA cm-2. Additionally, iohexol-intensified groundwater was used to compare the efficiency of anodes. The Ti/RuO2-TiO2 is also more efficient in removing the organic charge from the real water matrix (21.7% TOC) than the commercial anodes. Notably, the iohexol removal achieved is higher than all electrochemical treatments already reported using state-of-the-art non-active anodes in lower electrolysis time. Therefore, data from this study indicate that the electrochemical degradation of iohexol using the Ti/RuO2-TiO2 anode is efficient and has excellent cost-effectiveness; thus, it is a promising approach in the degradation of iohexol from wastewater. Furthermore, the Ti/RuO2-TiO2 active anode is competitive and can be an excellent option for treating effluents contaminated with recalcitrant organic compounds such as iohexol.

Pulp wash: a new source for production of ligninolytic enzymes and biomass and its toxicological evaluation after biological treatment.

Pulp wash was used as substrate for the activity of ligninolytic enzymes of the fungus Pleurotus sajor-caju. Activity of laccase (Lac) and manganese peroxidase (MnP) as well as fungal biomass occurred under four conditions: different pulp wash concentrations, pH variation at the optimal pulp wash concentration, different glucose concentrations, and different concentrations of ammonium nitrate. The best enzyme activity and biomass production were obtained with in natura pulp wash and pH corrected to 5.0 (4884 IU/L Lac; 82 IU/L MnP; 25 g/100 mL biomass). However, the addition of glucose and ammonium nitrate to the pulp wash was not necessary for increasing the enzyme activity and biomass production. Efficient removal of pulp wash chemical oxygen demand (99.66%) and biochemical oxygen demand (83.27%) occurred after the mycoremediation with P. sajor-caju in the optimized conditions. Lactuca sativa L. seeds germination bioassay showed a four-fold reduction in the residue toxicity (EC50 28.72%) after the treatment with the fungus. Our findings are consistent with the notion that pulp wash is an excellent substrate for inducing the activity of ligninolytic enzymes and producing fungal biomass, and that the biological treatment is efficient to reduce effluent toxicity.

Role of the antioxidant defense system during the production of lignocellulolytic enzymes by fungi.

Fungi are used for the production of several compounds and the efficiency of biotechnological processes is directly related to the metabolic activity of these microorganisms. The reactions catalyzed by lignocellulolytic enzymes are oxidative and generate reactive oxygen species (ROS). Excess of ROS can cause serious damages to cells, including cell death. Thus, the objective of this work was to evaluate the lignocellulolytic enzymes produced by Pleurotus sajor-caju CCB020, Phanerochaete chrysosporium ATCC 28326, Trichoderma reesei RUT-C30, and Aspergillus niger IZ-9 grown in sugarcane bagasse and two yeast extract (YE) concentrations and characterize the antioxidant defense system of fungal cells by the activities of superoxide dismutase (SOD) and catalase (CAT). Pleurotus sajor-caju exhibited the highest activities of laccase and peroxidase in sugarcane bagasse with 2.6 g of YE and an increased activity of manganese peroxidase in sugarcane bagasse with 1.3 g of YE was observed. However, P. chrysosporium showed the highest activities of exoglucanase and endoglucanase in sugarcane bagasse with 1.3 g of YE. Lipid peroxidation and variations in SOD and CAT activities were observed during the production of lignocellulolytic enzymes and depending on the YE concentrations. The antioxidant defense system was induced in response to the oxidative stress caused by imbalances between the production and the detoxification of ROS.