Lignocellulose-degrading fungi newly isolated from central Morocco are potent biocatalysts for olive pomace valorization.

The investigation of lignocellulolytic catalysts is an important feature to face the challenges of lignocellulosic biomass valorization. In central Morocco, fungi were isolated from decaying wood, soil, olive crushing by-products and their compost. One hundred fifty-five isolates were submitted to a selective screening, which served to distinguish 83% of lignocellulolytic isolates. Then, a collection of 56 fungi was subjected to morphological and molecular identification with the ITS5 and ITS4 primers. This approach showed that 45% of the fungal population belonged to the genus Penicillium, followed by Aspergillus 14%, and Fusarium 11%. Alternaria, Trichoderma, Paecilomyces, Cladosporium, Trichocladium, Circinella, and Doratomyces genera are founded with a minority occurrence. Finally, validation of the enzymatic profile was done for 20 isolates, by testing their enzymatic performance on a liquid medium in the presence of cellulose, lignin, and olive pomace. The maximum protein production of 788 µg ml-1 was reached by an Alternaria strain, which produced also 10.6 IU ml-1 of endoglucanase. Thus, a ß-glucosidase activity of 5.1 IU ml-1 was obtained by a Penicillium strain isolated from decaying wood. Regarding ligninolytic activities, olive pomace was the most suitable substrate to detect these activities. Decaying wood strains presented the most remarkable results with 1.1 IU ml-1, 0.7 IU ml-1 et 0.3 IU ml-1 for laccase, LiP and MnP, respectively. The use of the selected fungi and olive pomace as local biomass are important factors for the development of green processes targeting the valorization of this by-product into high-value molecules.

Lignocellulolytic Enzymes Production by Four Wild Filamentous Fungi for Olive Stones Valorization: Comparing Three Fermentation Regimens.

Lignocellulolytic enzymes play a crucial role in efficiently converting lignocellulose into valuable platform molecules in various industries. However, they are limited by their production yields, costs, and stability. Consequently, their production by producers adapted to local environments and the choice of low-cost raw materials can address these limitations. Due to the large amounts of olive stones (OS) generated in Morocco which are still undervalued, Penicillium crustosum, Fusarium nygamai, Trichoderma capillare, and Aspergillus calidoustus, are cultivated under different fermentation techniques using this by-product as a local lignocellulosic substrate. Based on a multilevel factorial design, their potential to produce lignocellulolytic enzymes during 15 days of dark incubation was evaluated. The results revealed that P. crustosum expressed a maximum total cellulase activity of 10.9 IU/ml under sequential fermentation (SF) and 3.6 IU/ml of ß-glucosidase activity under submerged fermentation (SmF). F. nygamai recorded the best laccase activity of 9 IU/ml under solid-state fermentation (SSF). Unlike T. capillare, SF was the inducive culture for the former activity with 7.6 IU/ml. A. calidoustus produced, respectively, 1,009 µg/ml of proteins and 11.5 IU/ml of endoglucanase activity as the best results achieved. Optimum cellulase production took place after the 5th day under SF, while ligninases occurred between the 9th and the 11th days under SSF. This study reports for the first time the lignocellulolytic activities of F. nygamai and A. calidoustus. Furthermore, it underlines the potential of the four fungi as biomass decomposers for environmentally-friendly applications, emphasizing the efficiency of OS as an inducing substrate for enzyme production.

Consolidated bioethanol production from olive mill waste: Wood-decay fungi from central Morocco as promising decomposition and fermentation biocatalysts.

Meknes region is a Moroccan olive-processing area generating high amounts of non-valorized Olive Mill Waste (OMW). Fungi are natural decomposers producing varied enzyme classes and effectively contributing to the carbon cycle. However, structural complexity of biomass and modest performances of wild fungi are major limits for local biorefineries. The objective of current research is to assess the ability of local fungi for bioethanol production from OMW using Consolidated Bioprocessing (CBP). This is done by characterizing lignocellulolytic potential of six wood-decay and compost-inhabiting ascomycetes and selecting potent fermentation biocatalysts. High and diversified activities were expressed by Fusarium solani and Fusarium oxysporum: 9.36 IU. mL-1 and 2.88 IU. mL-1 total cellulase activity, 0.54 IU. mL-1 and 0.57 IU. mL-1 laccase activity, respectively, and 8.43 IU. mL-1 lignin peroxidase activity for the latter. F. oxysporum had maximum bioethanol production and yield of 2.47 g.L-1 and 0.84 g.g-1, respectively, qualifying it as an important bio-agent for single-pot local biorefinery.