Lignocellulosic biomass is a significant source of sustainable fuel and high-value chemical production. However, due to the complex cross-linked three-dimensional network structure, lignin is highly rigid to degradation. In natural environments, the degradation is performed by wood-rotting fungi. The process is slow, and thus, the use of lignin degradation by fungi has not been regarded as a feasible technology in the industrial lignocellulose treatment. Fungi produce a wide variety of ligninolytic enzymes that can be directly introduced in industrial processing of lignocellulose. Within this study, screening of ligninolytic enzyme production using decolorization of ABTS and Azure B dyes was performed for 10 fungal strains with potentially high enzyme production abilities. In addition to standard screening methods, media containing lignin and hay biomass as carbon sources were used to determine the change in enzyme production depending on the substrate. All selected fungi demonstrated the ability to adapt to a carbon source limitation; however, four strains indicated the ability to secrete ligninolytic enzymes in all experimental conditions—Irpex lacteus, Pleurotus dryinus, Bjerkandera adusta, and Trametes versicolor—respectively displayed a 100%, 82.7%, 82.7%, and 55% oxidation of ABTS on lignin-containing media and 100%, 87.9%, 78%, and 70% oxidation of ABTS on hay-containing media after 168 h of incubation. As a result, the most potent strains of fungi were selected to produce lignocellulose-degrading enzymes and to demonstrate their potential application in biological lignocellulose pretreatment.

Lignocellulosic biomass can be used as a source for energy, fuel and valuable chemical production. From all available technologies, biological approaches have been recognized as the most environmentally friendly and sustainable ones. At the same time, high conversion costs, low efficiency and environmental issues still hinder the introduction of biological processes into industrial scale manufacturing. The aim of this study was to determine the most suitable enzyme cocktail recovery conditions from a biomass–fungal culture of the white-rot basidiomycete Irpex lacteus. Subsequent evaluation of the overall enzyme cocktail efficiency to release fermentable carbohydrates from biomass showed that prolonged fungal cultivation decreases the quality of the produced enzyme cocktail. At the same time, introduction of ultrasound pre-treatment during enzyme extraction improved the recovered enzyme cocktail efficiency in converting biomass to fermentable sugars, yielding up to 0.25 g of fermentable sugar per g dry hay biomass and up to 0.11 g per g dried straw or microalgae substrates. The results demonstrated that the production of lignocellulose-degrading enzymes from fungi is more sensitive than previously described, especially in terms of fungal growth, culture sterility and incubation conditions.

Tertiary wastewater treatment with microalgae incorporates environmental sustainability with future technologies and high exploitation costs. Despite the apparent ecological benefits of microalgae-assisted wastewater treatment/biomass-based resource production, technological improvements are still essential to compete with other technologies. Bio-flocculation instead of mechanical harvesting has been demonstrated as an alternative cost-effective approach. So far, mostly filamentous fungi of genus Aspergillus have been used for this purpose. Within this study, we demonstrate a novel approach of using white-rot fungi, with especially high potential of algae—Irpex lacteus complex that demonstrates efficiency with various microalgae species at a broad range of temperatures (5–20 °C) and various pH levels. Harvesting of microalgae from primary and secondary wastewater resulted in 73–93% removal efficiencies within the first 24 h and up to 95% after 48 h. The apparent reuse potential of the algae—I. lacteus pellets further complements the reduced operating costs and environmental sustainability of bio-flocculation technology.

The growing demand for novel enzyme producers to meet industrial and environmental needs has driven interest in lignocellulose-degrading fungi. In this study, lignocellulolytic enzyme production capabilities of environmental fungal isolates collected from boreal coniferous and nemoral summer green deciduous forests were investigated, using Congo Red, ABTS, and Azure B as indicators of cellulolytic and ligninolytic enzyme productions. Through qualitative and quantitative assays, the study aimed to identify promising species for lignocellulose-degrading enzyme secretion and assess their potential for biotechnological applications. Primary screening tests showed intensive enzyme secretion by certain isolates, particularly white rot fungi identified as Trametes pubescens and Cerrena unicolor. These fungi exhibited high efficiency in degrading Congo Red and Azure B. The isolates achieved up to a 93.30% decrease in Congo Red induced color intensity and over 78% decolorization of Azure B within 168 hours. Within 336 hours, these fungi reached nearly 99% removal of Congo Red and up to 99.79% decolorization of Azure B. Enzyme activity analysis confirmed the lignin-degrading capabilities of T. pubescens, which exhibited laccase activity exceeding 208 U/mL. Furthermore, Fomitopsis pinicola showed the highest cellulose-degrading potential among the studied fungi, achieving cellulase activity over 107 U/L during Congo Red decolorization. Previously undescribed enzyme-producing species, such as Peniophora cinerea, Phacidium subcorticalis, and Cladosporium pseudocladosporioides, also demonstrated promising lignocellulolytic enzyme production potential, achieving up to 98.65% and 99.80% decolorization of Congo Red and Azure B, respectively. The study demonstrates novel candidates for efficient lignocellulolytic enzyme production with broad biotechnological applications such as biomass conversion, wastewater treatment, textile dye and other complex chemical removal, and environmental remediation.

Wastewater treatment plants (WWTPs), particularly activated sludge systems, generate significant amounts of various types of waste, including screenings, primary sludge, and secondary sludge. While substantial research has been conducted on the recovery and valorization of sewage sludge, the treatment and utilization of screenings remain underexplored. In response, this study investigates the potential of white rot fungi to degrade cellulose-containing waste screened during the preliminary treatment and examines the production patterns of lignocellulolytic enzymes in the presence of this waste. The studied fungi exhibited variable enzymatic responses depending on the type of substrate, however, their adaptability highlighted the potential in fungal-mediated bioconversion processes. P. dryinus and T. versicolor were identified as strong and adaptive candidates for oxidative enzyme production, with P. dryinus showing laccase activity up to 1691.75 ± 12.22 U/mg and degrading 44.46% of carbohydrates in tested screenings. I. lacteus and B. adusta were predominantly observed in cellulolytic enzyme production, with B. adusta ensuring a 43.49% reduction in carbohydrate content of screenings. As a result of fungal cultivation in WWTP waste, the production potential of 34 to 46 kg of sugars per ton of screenings was determined. Therefore, the study presents a promising approach for the sustainable treatment of screenings and the development of waste management and resource recovery strategies for WWTP-derived waste.

The study investigates the application of white rot fungi for reactor-scale microalgae harvesting and explores the mechanisms underlying the algal-fungal interactions and their impact on biomass composition. Enzymatic analysis and microscopy revealed that the formation of algal-fungal complexes and successful harvesting are coupled with fungal cellulose-degrading enzyme production and hydrolytic processes of microalgae cells. Fluorescence intensity decreased by over 80 % in cells stained with Calcofluor-white after interaction with white rot fungi, indicating the reduction in cellulose content in microalgal cells caused by fungal enzymatic activity. These enzymes also caused significant cell damage and more than 50 % decrease in microalgae cell size. The presence of cellulolytic enzymes broadens the potential application of the resulting biomass in various biotechnological applications. Moreover, reactor-scale bioflocculation resulted in over 95 % T. obliquus and almost 85 % C. vulgaris harvesting efficiency from secondary wastewater within less than 24 h, demonstrating the method's scalability and industrial applicability.