We studied the fungal endophyte community of the leaves of Myracrodruon urundeuva in Brazil along with their potential to produce L-asparaginase. In total, 187 endophytes were isolated. The colonization rate of leaf fragments from caatinga (10.41 %) was lower than those from brejo de altitude (39.58 %). Sequences of ITS rDNA from all endophytes revealed relationships with Ascomycota (Botryosphaeriales, Chaetothyriales, Diaporthales, Eurotiales, Glomerellales, Hypocreales, and Pleosporales) and Basidiomycota (Polyporales). The most frequent endophytes were members of the genus Diaporthe. Talaromyces was an indicator genus for caatinga while Phyllosticta, Diaporthe, and Colletotrichum were for brejo de altitude. The composition of endophytic fungi in caatinga and brejo de altitude differed significantly, its richness and diversity (H' = 3.41) being significantly higher in brejo de altitude. Diaporthe sp. URM 7793 was the best producer of L-asparaginase (2.41 U/g), followed by Diaporthe sp. URM 7779 (2.00 U/g), Talaromyces sp. URM 7785 (1.91 U/g), and Diaporthe sp. URM 7792 (1.47 U/g). The composition of endophytic fungi assemblages is strongly influenced by the type of ecosystem. In its natural habitat, M. urundeuva harbors an important diversity of endophytes, which could be used to produce L-asparaginase.

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Acremonium sp. L1-4B isolated from lichen in Antarctica was used to produce extracellular proteases through submerged fermentation using cactus pear extract (Opuntia ficus-indica Mill.). A 2 3 factorial design was applied to optimize the protease production using three independent variables, namely temperature, pH and concentration of yeast extract, was also used a Central Composite Design (CCD) under Response Surface Methodology (RSM). All variables and interactions analyzed in the factorial design were significant or marginally significant, a Central Composite Design was developed, and the Response Surface Methodology towards the highest point it was established. The experimental model was validated under 14°C, pH 7.54, and 0.55% yeast extract, showing a protease activity of 447.65 7 2.6 U/mL by a prediction model of 445.48 U/mL. The enzyme showed a molecular weight of 59 kDa; it was inhibited in the presence of PMSF (serine protease); it presented optimal conditions at pH 8.0 and 50°C; it remained stable at pH in the 3.0-9.0 range and between 10 and 40°C; it showed a tolerance to 3000 mM NaCl as well as to surfactants, hydrogen peroxide and urea at 5%. This paper presents a proposal for an economically attractive production methodology using cactus pear as a primary source of carbon. In addition, the protease secreted by Acremonium sp. L1-4B presented a combination of biochemical characteristics that grants a promising variability of biotechnological applications.

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