RESUMO
The white rot basidiomycete Ganoderma lucidum was evaluated for its capability to tolerate and to degrade the herbicide diuron. Diuron at a subtoxic concentration was added at the start of the cultivation in glucose liquid stationary cultures. Under this condition diuron was a laccase inducer. Almost 50% of the initially present diuron was removed after 15 d of cultivation. Two diuron metabolites were found N'-(3,4-dichlorophenyl)-N-methylurea (DCPMU) and 3,4-dichlorophenylurea (DCPU). The addition of the cytochrome P450 inhibitors 1-aminobenzotriazole and piperonyl butoxide reduced significantly the capability of the fungus in degrading diuron. The activities of superoxide dismutase and catalase were significantly increased in the mycelial extracts by the presence of diuron. On the other hand, diuron did not cause any significant alteration in the levels of reactive oxygen species. Additionally, laccase could also degrade diuron in vitro and this degradation was increased by the addition of synthetic mediators, 3-ethylbenzthiazoline-6-sulphonic acid and acetylacetone. Significant reduction in the toxicity, as evaluated by the Lactuca sativa bioassay, was observed after G. lucidum treatment. In conclusion, G. lucidum is able to metabolize diuron by intra- and extracellular mechanisms, without the accumulation of toxic products.
Assuntos
Diurona/metabolismo , Farmacorresistência Fúngica , Herbicidas/metabolismo , Reishi/metabolismo , Biotransformação , Catalase/metabolismo , Diurona/farmacologia , Herbicidas/farmacologia , Lacase/metabolismo , Pentanonas/farmacologia , Butóxido de Piperonila/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Reishi/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Triazóis/farmacologiaRESUMO
In this work, a ß-glucosidase of Aspergillus awamori with a molecular weight of 180â¯kDa was produced in solid-state cultures using a mixture of pineapple crown leaves and wheat bran. Maximum production of the enzyme (820⯱â¯30â¯U/g substrate) was obtained after 8â¯days of culture at 28⯰C and initial moisture of 80%. The crude enzyme was efficiently immobilized on glutaraldehyde cross-linked commercial gelatin. Immobilization changed the kinetics of the enzyme, whose behavior could no longer be described by a saturation function of the Michaelis-Menten type. Comparative evaluation of the free and immobilized enzyme showed that the immobilized enzyme was more thermostable and less inhibited by glucose than the free form. In consequence of these properties, the immobilized enzyme was able to hydrolyze cellobiose more extensively. In association with Trichoderma reesei cellulase, the free and immobilized ß-glucosidase increased the liberation of glucose from cellulose 3- and 5-fold, respectively. Immobilization of the A. awamori ß-glucosidase on glutaraldehyde cross-linked commercial gelatin is an efficient and cheap method allowing the reuse of the enzyme by at least 10 times.
Assuntos
Aspergillus/enzimologia , Enzimas Imobilizadas/química , Gelatina/química , beta-Glucosidase/química , Celobiose/química , Celulose/química , Glucose/química , Hidrólise , Cinética , TemperaturaRESUMO
Yerba mate (Ilex paraguariensis) is a plant that grows naturally in South America. From its leaves and thin stems different kinds of beverages are prepared (chimarrão, tererê and tea mate), all of them rich in bioactive substances. The aim of this study was to evaluate the influence of in vitro gastrointestinal digestion and colonic fermentation on the stability of the polyphenols and on the antioxidant, antimicrobial and antitumoral activities of the yerba mate beverages. The phenolic chromatographic profile revealed that both the in vitro digestion and the colonic fermentation caused a pronounced decrease in 3,5-O-dicaffeoylquinic acid and 5-O-caffeoylquinic acid in the preparations. However, 3-O-caffeoylquinic acid, 4-O-caffeoylquinic acid and salvianolic acid I were only barely affected in all preparations. Despite the decrease in the phytochemicals content, yerba mate beverages maintain their functional properties such as antioxidant, antibacterial and antitumoral activities.
Assuntos
Fermentação , Ilex paraguariensis/metabolismo , Bebidas , Extratos VegetaisRESUMO
In this work, liquid nitrogen was used for the first time in the pretreatment of plant biomasses for purposes of enzymatic saccharification. After treatment (cryocrushing), the initial rates of the enzymatic hydrolysis of eucalyptus sawdust and rice hull were increased more than ten-fold. Cryocrushing did not modify significantly the contents of cellulose, hemicellulose and lignin in both eucalyptus sawdust and rice hulls. However, substantial disorganization of the lignocellulosic materials in consequence of the pretreatment could be observed by electron microscopy. Cryocrushing was highly efficient in improving the saccharification of the holocellulose component of the plant biomasses (from 4.3% to 54.1% for eucalyptus sawdust and from 3.9% to 40.6% for rice hull). It is important to emphasize that it consists in a simple operation with low requirements of water and chemicals, no corrosion, no release of products such as soluble phenolics, furfural and hydroxymethylfurfural and no waste generation.
Assuntos
Eucalyptus/química , Nitrogênio/química , Oryza/química , Biomassa , Celulose/química , Hidrólise , Lignina/análise , Nitrogênio/análise , Polissacarídeos/química , Madeira/químicaRESUMO
Pleurotus pulmonarius was cultivated on a corncob-based substrate for producing of mushrooms and for assessing the transformation of the lignocellulosics during the development of fungal biomass. Associated events, such as the release of relevant enzymes and the H2O2 generation, were also monitored. The peaks of laccase and catalase activities occurred at the 5th day and that of Mn peroxidase at the 30th day, simultaneously with a high activity of superoxide dismutase. Increase in the endocellulase and xylanase activities was observed after 10 days, with maximal activities achieved during the 20-30-day period. Maximal values of H2O2 were found after 10 days of cultivation. Electron microscopy and Fourier transform infrared (FTIR) spectroscopy showed strong alterations in the lignocellulosic fibers. The uncultivated and the cultivated substrates at different times were hydrolyzed with commercial cellulase and ß-glucosidase. The highest values of reducing sugars (110.5 ± 5.6 µmol/mL), being 65 % glucose, were obtained using the 20-day cultivated substrate. After the fruiting stage (first flush), enzymatic hydrolysis of the spent mushroom substrate (SMS) yielded 53.0 ± 2.8 and 77.5 ± 4.0 µmol/mL of glucose and total reducing sugars, respectively. Although the release of reducing sugars of the P. pulmonarius SMS was lower than that obtained after 20 days of cultivation, it was still 50 % higher than that obtained using the uncultured substrate. This observation, combined with the fact that SMS constitutes a residue generated as a by-product of the depletion of an agro-industrial residue, allows to conclude that this material offers an interesting economic perspective for the obtainment of cellulosic ethanol.