Evaluation of the Fermentative Capacity of Saccharomyces cerevisiae CAT-1 and BB9 Strains and Pichia kudriavzevii BB2 at Simulated Industrial Conditions.

The search for promising yeasts that surpass the fermentative capacity of commercial strains, such as Saccharomyces cerevisiae CAT-1, is of great importance for industrial ethanol processes in the world. Two yeasts, Pichia kudriavzevii BB2 and Saccharomyces cerevisiae BB9, were evaluated in comparison to the industrial yeast S. cerevisiae CAT-1. The objective was to evaluate the performance profile of the three studied strains in terms of growth, substrate consumption, and metabolite formation, aiming to determine their behaviour in different media and pH conditions. The results showed that under cultivation conditions simulating the medium used in the industrial process (must at 22° Brix at pH 3.0) the highest ethanol productivity was 0.41 g L-1 h-1 for S. cerevisiae CAT-1, compared to 0.11 g L-1 h-1 and 0.16 g L-1 h-1 for P. kudriavzevii and S. cerevisiae BB2, respectively. S. cerevisiae CAT-1 produced three times more ethanol in must at pH 3.0 (28.30 g L-1) and in mineral medium at pH 3.0 (29.17 g L-1) and 5.0 (30.70 g L-1) when compared to the value obtained in sugarcane must pH 3.0 (9.89 g L-1). It was concluded that S. cerevisiae CAT-1 was not limited by the variation in pH in the mineral medium due to its nutritional composition, guaranteeing better performance of the yeast even in the presence of stressors. Only S. cerevisiae CAT-1 expressed he constitutive invertase enzyme, which is responsible for hydrolysing the sucrose contained in the must.

Biochemical characterization and evaluation of invertases produced from Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa for the production of fructooligosaccharides.

Invertases are used for several purposes; one among these is the production of fructooligosaccharides. The aim of this study was to biochemically characterize invertase from industrial Saccharomyces cerevisiae CAT-1 and Rhodotorula mucilaginosa isolated from Cerrado soil. The optimum pH and temperature were 4.0 and 70 °C for Rhodotorula mucilaginosa invertase and 4.5 and 50 °C for Saccharomyces cerevisiae invertase. The pH and thermal stability from 3.0 to 10.5 and 75 °C for R. mucilaginosa invertase, respectively. The pH and thermal stability for S. cerevisiae CAT-1 invertase from 3.0 to 7.0, and 50 °C, respectively. Both enzymes showed good catalytic activity with 10% of ethanol in reaction mixture. The hydrolysis by invertases occurs predominantly when sucrose concentrations are ≤5%. On the other hand, the increase in the concentration of sucrose to levels above 10% results in the highest transferase activity, reaching about 13.3 g/L of nystose by S. cerevisiae invertase and 12.6 g/L by R. mucilaginosa invertase. The results demonstrate the high structural stability of the enzyme produced by R. mucilaginosa, which is an extremely interesting feature that would enable the application of this enzyme in industrial processes.

Catalytic Properties of Amylolytic Enzymes Produced by Gongronella butleri Using Agroindustrial Residues on Solid-State Fermentation.

Amylases catalyze the hydrolysis of starch, a vegetable polysaccharide abundant in nature. These enzymes can be utilized in the production of syrups, alcohol, detergent, pharmaceutical products, and animal feed formulations. The aim of this study was to optimize the production of amylases by the filamentous fungus Gongronella butleri by solid-state fermentation and to evaluate the catalytic properties of the obtained enzymatic extract. The highest amylase production, 63.25 U g-1 (or 6.32 U mL-1), was obtained by culturing the fungus in wheat bran with 55% of initial moisture, cultivated for 96 h at 25°C. The enzyme presented optimum activity at pH 5.0 and 55°C. The amylase produced was stable in a wide pH range (3.5-9.5) and maintained its catalytic activity for 1 h at 40°C. Furthermore, the enzymatic extract hydrolyzed starches from different vegetable sources, presenting predominant dextrinizing activity for all substrates evaluated. However, the presence of glucose was observed in a higher concentration during hydrolysis of corn starch, indicating the synergistic action of endo- and exoamylases, which enables the application of this enzymatic extract to produce syrups from different starch sources.

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.