A combined biological process was investigated for effective textile wastewater treatment. The process consisted of a first step performed by selected fungal biomasses, mainly devoted to the effluent decolourisation, and of a subsequent stage by means of activated sludge, in order to reduce the remaining COD and toxicity. In particular, the treatment with Trametes pubescens MUT 2400, selected over nine strains, achieved very good results in respect to all parameters. The final scale-up phase in a moving bed bioreactor with the supported biomass of the fungus allowed to verify the effectiveness of the treatment with high volumes. Despite promising results, further steps must be taken in order to optimize the use of these biomasses for a full exploitation of their oxidative potential in textile wastewater treatment.

11Bioeconomy stresses the need of green processes promoting the development of 12 new methods for biocatalyzed alkene reductions. 13A functional screening of 28 fungi belonging to Ascomycota, Basidiomycota 14 and Zygomycota isolated from different habitats was performed. Their capability to 15 reduce C=C double bonds was evaluated towards three substrates (cyclohexenone, α-16 methylnitrostyrene and α-methylcinnamaldehyde) with different electron-withdrawing 17 groups, i.e., ketone, nitro and aldehyde, respectively. 18Almost all the fungi showed this reducing activity. Noteworthy Gliomastix 19 masseei, Mucor circinelloides and Mucor plumbeus resulted very versatile and 20 effective, being able to reduce all the model substrates quickly and with high yields. 21 2

Autochthonous fungi from contaminated wastewater are potential successful agents bioremediation thanks to their adaptation to pollutant toxicity and to competition with other microorganisms present in wastewater treatment plant. Biological treatment by means of selected fungal strains could be a potential tool to integrate the leachate depuration process, thanks to their fungal extracellular enzymes with non-selective catalytical activity. In the present work, the treatability of two real samples (a crude landfill leachate and the effluent coming from a traditional wastewater treatment plant) was investigated in decolorization experiments with fungal biomasses. Five autochthonous fungi, Penicillium brevicompactum MUT 793, Pseudallescheria boydii MUT 721, P. boydii MUT 1269, Phanerochaete sanguinea MUT 1284, and Flammulina velutipes MUT 1275, were selected in a previous miniaturized decolorization screening. Their effectiveness in terms of decolorization, enzymatic activity (laccases and peroxidases), biomass growth and ecotoxicity removal was compared with that of five allochthonous fungal strains, Pleurotus ostreatus MUT 2976, Porostereum spadiceum MUT 1585, Trametes pubescens MUT 2400, Bjerkandera adusta MUT 3060 and B. adusta MUT 2295, selected for their well known capability to degrade recalcitrant pollutants. Moreover, the effect of biomass immobilization on polyurethane foam (PUF) cube was assessed. The best decolorization (60%) was achieved by P. spadiceum MUT 1585, P. boydii MUT 721 and MUT 1269. In the first case, the DP was achieved gradually, suggesting a biodegradation process with the involvement of peroxidases. On the contrary, the two autochthonous fungi seem to bioremediate the effluent mainly by biosorption, with the abatement of the toxicity (up to 100%). The biomass immobilization enhanced enzymatic activity, but not the DP. Moreover, it limited the biomass growth for the fast growing fungi, MUT 721 and MUT 1269. In conclusion, robust and versatile strains coming from well-characterized collections of microorganisms can obtain excellent results comparing and even exceeding the bioremediation yields of strains already adapted to pollutants.

The reduction of C=C double bond, a key reaction in organic synthesis, is mostly achieved by traditional chemical methods. Therefore, the search for enzymes capable of performing this reaction is rapidly increasing. Old Yellow Enzymes (OYEs) are flavin-dependent oxidoreductases, initially isolated from Saccharomyces pastorianus. In this study, the presence and activation of putative OYE enzymes was investigated in the filamentous fungus Mucor circinelloides, which was previously found to mediate C=C reduction. Following an in silico approach, using S. pastorianus OYE1 amminoacidic sequence as template, ten putative genes were identified in the genome of M. circinelloides. A phylogenetic analysis revealed a high homology of McOYE1-9 with OYE1-like proteins while McOYE10 showed similarity with thermophilic-like OYEs. The activation of mcoyes was evaluated during the transformation of three different model substrates. Cyclohexenone, α-methylcinnamaldehyde and methyl cinnamate were completely reduced in few hours and the induction of gene expression, assessed by qRT-PCR, was generally fast, suggesting a substrate-dependent activation. Eight genes were activated in the tested conditions suggesting that they may encode for active OYEs. Their expression over time correlated with C=C double bond reduction.

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