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.
Profiling microorganisms with potential for amylase production in low cost culture media has been widely recognized due to its broad applicability. The aim of this study was to select yeast strains with potential to produce amylolytic enzymes by solid state fermentation. Fifty-four (54) strains were assessed and three exhibited ability to produce amylases: Candida parapsilosis with 14.68 U/mL (146.8 U/g substrate); Rhodotorula mucilaginosa with 25.0 U/mL (250 U/g substrate), and Candida glabrata with 25.39 U/mL (253.9 U/g substrate), in solid state fermentation, for 120 h at 28°C, using wheat bran with 70% moisture. The enzymes exhibited maximum activity at a pH of 7.0 and at 60°C. Amylases demonstrated satisfactory structural stability, maintaining their catalytic activity after 1 h at 50°C. All enzymes were ethanol tolerant and retained more than 70% of their original activities in 15% ethanol solution. Corn starch was efficiently hydrolyzed by enzymes and the extracts produced by C. parapsilosis and C. glabrata exhibited dextrinizing activity, while those produced by R. mucilaginosa exhibited saccharifying activity.
ABSTRACT: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 3.0-11.0. 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.
Microbial β-glucosidases can be used in several industrial processes, including production of biofuels, functional foods, juices, and beverages. In the present work, production of β-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 β-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. β-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 noncompetitive inhibition. In the presence of up to 10% ethanol, β-glucosidase maintained its catalytic activity. In addition to β-glucosidase, the enzymatic extract showed activity of 36 U/g for endoglucanase, 256.2 U/g for xylanase, and 18.2 U/g for β-xylosidase. The results allow to conclude that the fungus T. crustaceus has considerable potential for production of β-glucosidase and xylanase when cultivated in agroindustrial residues, thereby reducing the cost of these biocatalysts.
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