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The strategy to reuse the mycelium grown in solid-state cultivation for enzyme production was developed in this work. Also, the nutrients and syringe-type bioreactor necessary for the enzymatic production have also been tested to make the process more sustainable and less onerous. For the best condition of supplementation in the mycelia turnover system, the enzymatic activities over filter paper were 5 times and 2 times higher than the ones found in the conventional production by Penicillium echinulatum S1M29 and Trichoderma reesei RUT C30, respectively. A xylanase activity of 1303 U/g by P. echinulatum S1M29 was reported in this work after the four cycles of extraction, showing the efficiency of this novel strategy for this fungus. The reuse of the mycelium in solid-state cultivation proved to be a promising strategy to increase production and productivity and reduce the amounts of inputs used in enzyme production processes.

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