Role of In-House Enzymatic Cocktails from Endophytic Fungi in the Saccharification of Corn Wastes Towards a Sustainable and Integrated Biorefinery Approach

The valorization of agri-food wastes can provide value-added products, enzymes and biofuels. For the second-generation ethanol (2G) production, pulps rich in cellulose are desirable in order to release fermentable sugars. This study investigated the homemade biosynthesis of cellulases and hemicellulases via solid-state fermentation (SSF) using sugarcane bagasse (SB) and wheat bran (WB) for the growth of endophytic fungi (Beauveria bassiana, Trichoderma asperellum, Metarhizium anisopliae and Pochonia chlamydosporia). Cocktails with high enzymatic levels were obtained, with an emphasis for M. anisopliae in the production of β-glucosidase (83.61 U/g after 288 h) and T. asperellum for xylanase (785.50 U/g after 144 h). This novel M. anisopliae β-glucosidase demonstrated acidophile and thermotolerant properties (optimum activity at pH 5.5 and 60 °C and stability in a wide pH range and up to 60 °C), which are suitable for lignocellulose saccharifications. Hence, the M. anisopliae multi-enzyme blend was selected for the hydrolysis of raw and organosolv-pretreated corn straw (CS) and corncob (CC) using 100 CBU/g cellulose. After the ethanol/water (1:1) pretreatment, solid fractions rich in cellulose (55.27 in CC and 50.70% in CS) and with low concentrations of hemicellulose and lignin were found. Pretreated CC and CS hydrolysates reached a maximum TRS release of 12.48 and 13.68 g/L, with increments of 100.80 and 73.82% in comparison to untreated biomass, respectively, emphasizing the fundamental role of a pretreatment in bioconversions. This is the first report on β-glucosidase biosynthesis using M. anisopliae and its use in biomass hydrolysis. These findings demonstrated a closed-loop strategy for internal enzyme biosynthesis integrated to reducing sugar release which would be applied for further usage in biorefineries.