Leonardo Casieri

Decolorization and detoxication of reactive industrial dyes by immobilized fungi Trametes pubescens and Pleurotus ostreatus

Trametes pubescens and Pleurotus ostreatus, immobilized on polyurethane foam cubes in bioreactors, were used to decolorize three industrial and model dyes at concentrations of 200, 1000 and 2000 ppm. Five sequential cycles were run for each dye and fungus. The activity of laccase, Mn-dependent and independent peroxidases, lignin peroxidase, and aryl-alcohol oxidase were daily monitored during the cycles and the toxicity of media containing 1000 and 2000 ppm of each dye was assessed by the Lemna minor (duckweed) ecotoxicity test. Both fungi were able to efficiently decolorize all dyes even at the highest concentration, and the duckweed test showed a significant reduction (p ≤ 0.05) of the toxicity after the decolorization treatment. T. pubescens enzyme activities varied greatly and no clear correlation between decolorization and enzyme activity was observed, while P. ostreatus showed constantly a high laccase activity during decolorization cycles. T. pubescens showed better decolorization and detoxication capability (compared to the better known P. ostreatus). As wide differences in enzyme activity of the individual strains were observed, the strong decolorization obtained with the two fungi suggested that different dye decolorization mechanisms might be involved.

Survey of ectomycorrhizal, litter-degrading, and wood-degrading Basidiomycetes for dye decolorization and ligninolytic enzyme activity

Basidiomycetes are essential in forest ecology, being deeply involved in wood and litter decomposition, humification, and mineralization of soil organic matter. The fungal oxidoreductases involved in these processes are today the focus of much attention with a view to their applications. The ecological role and potential biotechnological applications of 300 isolates of Basidiomycetes were assessed, taking into account the degradation of model dyes in different culture conditions and the production of oxidoreductase enzymes. The tested isolates belong to different ecophysiological groups (wood-degrading, litter-degrading, ectomycorrhizal, and coprophilous fungi) and represent a broad systematic and functional biodiversity among Basidiomycetes occurring in deciduous and evergreen forests of northwest Italy (Piedmont Region). The high number of species tested and the use of different culture conditions allowed the investigation of the degradation activity of several novel species, neglected to date. Oxidative enzyme activities varied widely among all ecophysiological groups and laccases were the most commonly detected enzymes. A large number of isolates (86%), belonging to all ecophysiological groups, were found to be active against at least one model dye; the wood-degrading fungi represented the most efficient group. Noteworthily, also some isolates of litter-degrading and ectomycorrhizal fungi achieved good decolorization yield. The 25 best isolates were then tested against nine industrial dyes commonly employed in textile industries. Three isolates of Bjerkandera adusta efficiently decolorized the dyes on all media and can be considered important candidates for application in textile wastewater treatment.

Interactions between engineered tomato plants expressing antifungal enzymes and nontarget fungi in the rhizosphere and phyllosphere

The introduction of genetically modified (GM) plants in agroecosystems raises concern about possible effects on nontarget species. The impact of a tomato line transformed for constitutive expression of tobacco beta-1,3-glucanase and chitinase on indigenous nonpathogenic fungi was investigated. In greenhouse experiments, no significant differences were found in the colonization by arbuscular mycorrhizal fungi. Diversity indices computed from over 20 500 colonies of culturable rhizosphere and phyllosphere saprotrophic microfungi, assigned to 165 species (plus > 80 sterile morphotypes), showed no significant differences between GM and wild-type plants. Differences were found by discriminant analysis in both the rhizosphere and the phyllosphere, but such effects were minor compared with those linked to different plant growth stages.

Basidiomycetes from Compost and their Dye Degradation and Enzyme Activities

The degree of decolorization of Poly R-478, a highly recalcitrant anthraquinone dye, by three basidiomycetes belonging to Polyporales isolated from compost was investigated together with the enzymes involved. Decolorization tests in two liquid cultures, one with a simple mycelium inoculum, the other with an inoculum of mycelium grown on straw, resulted in 70% and 87% decolorization respectively in 7 days. However, the efficiency did not increase significantly in the presence of the lignocellulose substrate. The three strains produced laccase and/or manganese peroxidase activity during the decolorization, whereas lignin peroxidase activity was not observed. Previous growth on straw enhanced the synthesis of ligninolytic enzymes, though there was no correlation between enzyme activity and decolorization. The three fungi can be proposed as promising candidates for the treatment of colored industrial effluents and probably for soils contaminated by complex polymers, such as polycyclic aromatic hydrocarbons.