ß-glucosidase from thermophilic fungus Thermoascus crustaceus: production and industrial potential.

Microbial ß-glucosidases can be used in several industrial processes, including production of biofuels, functional foods, juices, and beverages. In the present work, production of ß-glucosidase by solid state cultivation of the fungus Thermoascus crustaceus in a low-cost cultivation medium (comprising agroindustrial residues) was evaluated. The highest production of ß-glucosidase, about 415.1 U/g substrate (or 41.51 U/mL), was obtained by cultivating the fungus in wheat bran with 70% humidity, during 96 h at 40°C. The enzymatic activity was optimum at pH 4.5 and 65°C. ß-Glucosidase maintained its catalytic activity when incubated at a pH range of 4.0-8.0 and temperature of 30-55°C. The enzyme was strongly inhibited by glucose; even when the substrate and glucose concentrations were equal, the inhibition was not reversed, suggesting a non-competitive inhibition. In the presence of up to 10% ethanol, ß-glucosidase maintained its catalytic activity. In addition to ß-glucosidase, the enzymatic extract showed activity of 36 U/g for endoglucanase, 256.2 U/g for xylanase, and 18.2 U/g for ß-xylosidase. The results allow to conclude that the fungus T. crustaceus has considerable potential for production of ß-glucosidase and xylanase when cultivated in agroindustrial residues, thereby reducing the cost of these biocatalysts.

Production and Catalytic Properties of Amylases from Lichtheimia ramosa and Thermoascus aurantiacus by Solid-State Fermentation.

The present study compared the production and the catalytic properties of amylolytic enzymes obtained from the fungi Lichtheimia ramosa (mesophilic) and Thermoascus aurantiacus (thermophilic). The highest amylase production in both fungi was observed in wheat bran supplemented with nutrient solution (pH 4.0) after 96 hours of cultivation, reaching 417.2 U/g of dry substrate (or 41.72 U/mL) and 144.5 U/g of dry substrate (or 14.45 U/mL) for L. ramosa and T. aurantiacus, respectively. The enzymes showed higher catalytic activity at pH 6.0 at 60°C. The amylases produced by L. ramosa and T. aurantiacus were stable between pH 3.5-10.5 and pH 4.5-9.5, respectively. The amylase of L. ramosa was stable at 55°C after 1 hour of incubation, whereas that of T. aurantiacus maintained 60% of its original activity under the same conditions. Both enzymes were active in the presence of ethanol. The enzymes hydrolyzed starch from different sources, with the best results obtained with corn starch. The enzymatic complex produced by L. ramosa showed dextrinizing and saccharifying potential. The enzymatic extract produced by the fungus T. aurantiacus presented only saccharifying potential, releasing glucose monomers as the main hydrolysis product.

Production of xylanase by a new strain of Thermoascus aurantiacus: obtainment of enzymatic extract with reduced cellulolytic activity for application in pulp and paper industries / Produção de xilanase por nova linhagem de Thermoascus aurantiacus: obtenção de extrato enzimático com reduzida atividade celulolítica para aplicação em indústria de papel e celulose

Xylanases are useful in several industrial segments, including pulp and paper bleaching, animal feed, and bread-making processes. However, the industrial use of these enzymes is closely related to its production cost and its catalytic properties. The process of solid state fermentation enables the use of agro-industrial residues as substrates for microbial cultivation and enzymes production, reducing costs. In the present study, different cultivation parameters were evaluated for the xylanase production by the thermophilic fungus Thermoascus aurantiacus, by solid state fermentation, using agro-industrial residues as substrates. High production of xylanase (1701.9 U g-1 of dry substrate) was obtained using wheat bran containing 65% of initial moisture, at 120 h of cultivation, and 45°C. The xylanase showed optimal activity at pH 5.0 and 75°C; its stability was maintained at pH 3.0­11.0. The enzyme retained its catalytic potential after 1 h, at 75°C. The enzymatic extract produced under optimized conditions showed reduced activities of endoglucanase and FPase. Our results, including the xylanase production by T. aurantiacus in low-cost cultivation medium, high structural stability of the enzyme, and reduced cellulolytic activity, encourage the application of this enzymatic extract in pulp and paper bleaching processes.

As xilanases apresentam aplicabilidade em diferentes segmentos industriais, como: branqueamento de papel e celulose, ração animal e panificação. No entanto, a utilização industrial dessas enzimas está intimamente relacionada com seu custo de produção e suas propriedades catalíticas. O processo de fermentação em estado sólido possibilita o uso de resíduos agroindustriais como substratos, para o cultivo microbiano e produção de enzimas, reduzindo o custo da produção enzimática. No presente trabalho, diferentes parâmetros de cultivo foram avaliados para produção de xilanase por cultivo em estado sólido do fungo termófilo Thermoascus aurantiacus, utilizando resíduos agroindustriais como substratos. A maior produção de xilanase, 1701,9 U g-1 de substrato seco, foi obtida no cultivo em farelo de trigo, contendo 65% de umidade inicial, em 120 horas de cultivo a 45°C. A xilanase produzida apresentou atividade ótima em pH 5,0 a 75°C, mantendo sua estabilidade em pH 3,0 a 11,0. A enzima manteve seu potencial catalítico após 1 h a 75°C. O extrato enzimático produzido nas condições otimizadas apresentou reduzida atividade de endoglucanase e FPase. Os resultados obtidos no presente trabalho (produção de xilanase pelo fungo em meios de cultivo de baixo custo, elevada estabilidade estrutural da enzima e reduzida atividade celulolítica) estimulam a aplicação desse complexo enzimático em processos de branqueamento de papel e celulose.