Marcelo Franco

Optimization of productions of cellulolytic enzymes by Aspergillus niger using residue of mango a substrate

The present paper analyses the effects of water activity (0.88, 0.94 and 0.97) and of fermentation time (24, 48, 72, 96 and 120 hours) on the kinetic activity of enzymes cellulolytic, produced during the solid state fermentation of waste from the improvement of mango, with the aid of fungus species Aspergillus niger. Solid state fermentation was carried out at 35°C inside a bacteriological incubator. The statistical results indicated that the best activity for enzyme CMCase was 7.26U g-1 after 74.51 hours of fermentation, whereas for enzyme FPase was 2.55U g-1 after 98.52 hours, both presenting best results in approximately 0.928 of water activity. Pareto charts have showed that fermentation time has greater effect over the activity of enzyme CMCase, while the water activity variable has greater effect over enzyme FPase activity. During fermentation the fungus synthesized the enzymes without the need of inductors other than mango residue and water.

Application of Response Surface Methodology for Producing Cellulolytic Enzymes by Solid-state Fermentation from the Puple Mombin (Spondias purpurea L.) Residue

This study aimed at demonstrating the effects of fermentation time (24, 48, 72, 96, and 120 h) and water activity (0.943, 0.970, and 0.985) on the production of cellulolytic enzymes by solid-state fermentation of purple mombin (Spondias purpurea L.) residue using Aspergillus niger. The fermentation was carried out at 35°C and the enzyme production was measured as endoglucanase and total cellulose activities. The optimum condition for endoglucanase was water activity 0.974 and 93.8 h of fermentation, reaching a production of 3.21 U/g of residue; whereas for total cellulase it was 0.958 and 79.4 h achieving 12.1 U/g of residue. Fermentation time had a greater effect on the endoglucanase activity, while water activity had a more significant influence on the total cellulase activity. Endoglucanase had optimum activity at temperature of 50°C and pH 5.0. Although cellulase total optimum activity was also at pH 5.0, the maximum activity was at 60°C.

Enzymatic saccharification of lignocellulosic residues using cellulolytic enzyme extract produced by Penicillium roqueforti ATCC 10110 cultivated on residue of yellow mombin fruit

The aim of this work was to enzymatic saccharification of food waste was performed by crude enzymatic cellulolytic extract produced by P. roqueforti cultivated in yellow mombin residue. The best yield of reducing sugars (259.45mgg-1) was achieved with sugarcane bagasse after 4h; the hydrolysis of corn cob, rice husk and peanut hull resulted in yields around 128-180mgg-1. The addition of 10mmolL-1 of Mn2+ potentiated the saccharification of sugarcane bagasse, in about 86%. The temperature and substrate (sugarcane bagasse) concentration parameters were optimized using a Doehlert Design and, a maximum sugar yield of 662.34±26.72mgg-1 was achieved at 62.40°C, 0.22% (w/v) of substrate, with the addition of Mn2+. Sugar yield was significantly high when compared to previous studies available in scientific literature, suggesting the use of crude cellulolytic supplemented with Mn2+ an alternative and promising process for saccharification of sugarcane bagasse.

Prickly palm cactus husk as a raw material for production of ligninolytic enzymes by Aspergillus niger

Prickly palm cactus husk was used as a solid-state fermentation support-substrate for production of the ligninolytic enzymes laccase, peroxide manganese, and lignin peroxidase by Aspergillus niger. Effects of water activity, temperature, and fermentation time on enzymatic production were evaluated using a central composite rotatable design. Response surface methodology revealed that maximum enzyme production was achieved at 73.38 h of fermentation, a water activity of 0.87 Aw, at 28.74°C for laccase, at 65.33 h, 0.89 Aw, and 28.96°C for lignin peroxidase, and at 70.44 h, 0.91 Aw, and 28.84°C for manganese peroxidase. Optimized enzyme production was 9,023.67 UI/L for laccase, 2,234.75 UI/L for lignin peroxidase, and 8,534.81 UI/L for manganese peroxidase. Thermostability and pH stability were observed for all enzymes. Enzymatic deactivation kinetic experiments indicated that enzymes remained active after freezing of crude extracts.

Statistical Optimization of Culture Conditions and Characterization for Ligninolytic Enzymes Produced from Rhizopus Sp. Using Prickly Palm Cactus Husk

Dry prickly palm cactus (Nopalea cochenillifera) husk was investigated as a substrate for Rhizopus sp. cultivation in the solid state, aiming at the production of laccase (Lac), lignin peroxidase (LiP), and manganese peroxidase (MnP). The optimization of fermentation was evaluated by an experimental design and it was obtained, for each enzyme, maximum productivities (U g−1 h−1) of: 0.085 ± 0.02 (MnP), 0.066 ± 0.001 (LiP), and 0.023 ± 2.3.10−4 (Lac), at the conditions of 10 g of substrate, 72 h of fermentation, aw = 0.865, and 30°C. The enzymes thermal and pH stabilities were evaluated and it was observed better results at temperatures no higher than 60°C and pH of 5.0; in addition, the storage of these enzymes was better at − 25°C than at 4°C. Since the prickly palm cactus is an agricultural substrate and specially because of its low cost, it is important to propose different applications for it as, for example, an alternative substrate for biotechnological processes.

Comparison between the univariate and multivariate analysis on the partial characterization of the endoglucanase produced in the solid state fermentation by Aspergillus oryzae ATCC 10124

ABSTRACT Endoglucanase production by Aspergillus oryzae ATCC 10124 cultivated in rice husks or peanut shells was optimized by experimental design as a function of humidity, time, and temperature. The optimum temperature for the endoglucanase activity was estimated by a univariate analysis (one factor at the time) as 50°C (rice husks) and 60°C (peanut shells), however, by a multivariate analysis (synergism of factors), it was determined a different temperature (56°C) for endoglucanase from peanut shells. For the optimum pH, values determined by univariate and multivariate analysis were 5 and 5.2 (rice husk) and 5 and 7.6 (peanut shells). In addition, the best half-lives were observed at 50°C as 22.8 hr (rice husks) and 7.3 hr (peanut shells), also, 80% of residual activities was obtained between 30 and 50°C for both substrates, and the pH stability was improved at 5–7 (rice hulls) and 6–9 (peanut shells). Both endoglucanases obtained presented different characteristics as a result of the versatility of fungi in different substrates.

Production of lipase from Penicillium sp. using waste oils and Nopalea cochenillifera

ABSTRACT The present communication deals with the production of lipase from Penicillium sp. using waste oils and palm cactus (Nopalea cochenillifera) especially as nutrient source of low cost. Two different waste oils were tested: waste frying oil from an industrial kitchen and waste lubricating oil (WLO) from a gas station. Using Doehlert experimental design and response surface methodology, the optimum conditions for lipase production were 96 h fermentation, WLO as the inductor, with specific activity of 0.22 UA mg−1. The enzyme was able to remain with more than 58% of its original activity until 30 min at 60°C. The kinetic constants were Km = 9.93 mM and Vmax = 2.58 UA min−1 using p-nitrophenyl palmitate (p-NPP) as substrate. Results showed that Penicillium sp. was able to produce lipase from waste oils using N. cochenillifera, thus having biotechnological potential in waste oil biotransformation.

Aspergellus niger como produtor de enzimas celuloliticas a partir farelo de cacau (Theobroma cacao)

The aim of this work was to study the application of the fungus Aspergillus niger as a producer of the cellulolytic enzymes CMCase, FPase and Xylanase by solid-state fermentation of cocoa (Theobroma cacao). We evaluated the effect of fermentation time (24, 72, and 120 hours) and water activity (0.963, 0.976 and 0.983) on the production of enzymes. Fermentations were performed at 30o C in a bacteriological incubator. The optimization of ideal conditions for enzyme production was carried out using the response surface methodology (RSM). Statistically, the best activity obtained for CMCase was 14.18 U/mL at aw 0.972 and 70.07 hours fermentation, for FPase it was 7.51 U/mL at 0.974 aw and 80.56 hours, while for Xylanase was 11.86 U/mL at aw 0.971 and 64.24 hours.

Effect of the solid state fermentation of cocoa shell on the secondary metabolites, antioxidant activity, and fatty acids

Abstract During cocoa (Theobroma cacao L.) processing, the accumulated cocoa shell can be used for bioconversion to obtain valuable compounds. Here, we evaluate the effect of solid-state fermentation of cacao flour with Penicillium roqueforti on secondary metabolite composition, phenol, carotenoid, anthocyanin, flavonol, and fatty acids contents, and antioxidant activity. We found that the total concentrations of anthocyanins and flavonols did not change significantly after fermentation and the phenolic compound and total carotenoid concentrations were higher. The fermentation process produced an increase in saponin concentration and antioxidant activity, as well as significant changes in the levels of oleic, linoleic, gamma-linolenic, and saturated fatty acids. Based on our findings, we propose that the reuse of food residues through solid state fermentation is viable and useful.

Improvement in menthol extraction of fresh leaves of Mentha arvensis by the application of multi-enzymatic extract of Aspergillus niger

Abstract The objective of this study was to optimize the enzymatic pretreatment process of extracting menthol, a major component of the essential oil stored in the trichomes of fresh leaves of Mentha arvensis, by the action of the crude enzymatic extract previously produced by the Aspergillus niger fungus cultivated in cactus pear. Under the experimental conditions of 42 °C, 141 min and 58 mL of enzyme in 442 mL of water, there was an increase of 186.63% in the hydrodistillation yield, which represents an acceptable positive deviation of 12.54% compared to the predicted value of the model obtained by experimental design. Trichome rupture can be confirmed by scanning electron microscopy (SEM) and the chemical structure of the obtained menthol was not modified according to gas chromatography and infrared spectroscopy. The enzymatic pretreatment applied before the hydrodistillation step was successful and proves the potential of the association of biotechnological techniques with a physicochemical process.