Coupling chemical oxidation processes and Leptosphaerulina sp. myco-remediation to enhance the removal of recalcitrant organic pollutants in aqueous systems.

This research evaluated for the first time, the coupling of chemical oxidation processes with Leptosphaerulina sp. (a Colombian fungus), to degrade a refractory pollutant. For such purpose, a model contaminant (crystal violet, CV) was considered. Initially, the pollutant, at high concentrations (i.e., 200 and 50 mg L-1), was submitted to the fungus action. However, the CV inhibited the growth and enzymatic production of the fungus. Then, three chemical oxidation processes: TiO2-photocatalysis, sonochemistry, or electrochemistry (with a Ti/IrO2 anode in sodium chloride) were used as treatments previous to the myco-remediation. These oxidative treatments led to the pollutant degradation (~100%) by the action of radicals or active chlorine species, but they showed low mineralization. Indeed, the total organic carbon removal (TOC) was 54, ~15, and 31% to TiO2-photocatalysis (after 12 h), sonochemistry (after 12 h), and electrochemistry (after 1.33 h), respectively. Thus, the resultant solutions from the chemical oxidations were submitted to the action of Leptosphaerulina sp. (this time effective fungus growth and enzymes production were observed). It was found that the TOC removals by the fungus were 87, 84, and 83% for solutions pre-treated by TiO2-photocatalysis (12 h), sonochemical (12 h), and electrochemical (1.33 h) treatments, respectively. Regarding the enzymatic production, TiO2-photocatalysis/Leptosphaerulina sp., ultrasonication/Leptosphaerulina sp., and electrochemical oxidation/Leptosphaerulina sp. combinations reached the highest activities of laccase (0.6 U mg-1, at day 15), manganese peroxidase (1.35 U mg-1, at day 7) and versatile peroxidase (1.72 U mg-1, at day 15), respectively. The results from this work evidence feasibility of the pre-treatment with chemical oxidation processes as a strategy to enhance Leptosphaerulina sp. action toward recalcitrant organic pollutants (as CV) in water.

Enhancement of ligninolytic enzymes production and decolourising activity in Leptosphaerulina sp. by co-cultivation with Trichoderma viride and Aspergillus terreus.

This work investigated fungal co-culture as inducer of ligninolytic enzymes and decolourising activity in the Colombian strain Leptosphaerulina sp., an ascomycete white-rot fungus isolated from lignocellulosic material. Aspergillus niger, Aspergillus fumigatus, Aspergillus terreus, Trichoderma viride, Fusarium sp. and Penicillium chrysogenum were tested as Leptosphaerulina sp. inducers. The best fungal combinations in terms of enzyme production, fungal growth and decolourising activity were selected from solid media experiments. Response surface methodology (RSM) was utilised to optimise enzyme production and decolourising activity in liquid media. Solid media assays evidenced T. viride and A. terreus as the best Leptosphaerulina sp. inducers. The RSM identified a triple co-culture inoculated with T. viride (1000 µL) and A. terreus (1000 µL) into a 7-day culture of Leptosphaerulina sp. as the best treatment. This triple combination significantly improved ligninolytic enzymes production and Reactive Black 5 dye removal when compared to the Leptosphaerulina sp. monoculture and previously used chemical inducers. These results demonstrated the potential of fungal co-culture as an environmentally-friendly method to enhance Leptosphaerulina sp. enzymes production and decolourising activity.