Tannase is an enzyme that hydrolyzes esters and lateral bonds of tannins, such as tannic acid, releasing glucose and gallic acid and stands out in the clarification of wines and juices. Fungi of the genera Aspergillus and Penicillium are excellent producers of this enzyme. The search for fungi that produce high levels of tannase as well as new substrates for the enzyme production by the SSF is required. The objectives of this study were to evaluate the production of tannase by Aspergillus and Penicillium species through SSF using leaves and agroindustrial waste barbados cherry and mangaba fruit as substrate, select the best producer, optimize production, characterize the crude enzyme extract, and apply it the clarification of grape juice. Selecting the best producer was performed by planning Placket-Burman and RSM. P. montanense showed highest activity with 41.64 U/mL after 72 h of fermentation residue using barbados cherry, with 3.5% tannic acid and 70% moisture. The enzyme showed the highest activity at pH 9.0 and 50°C. The tannase of P. montanense was stable over a wide pH range and temperature and, when applied to grape juice, showed higher efficiency by reducing 46% of the tannin content after incubation 120 m.
ABSTRACT:The Caatinga biome features an exclusive endemic biodiversity, and is characterized by the presence of xerophytic, deciduous vegetation, high temperatures, and low rainfall. This important park has undergone anthropization, especially through extraction of firewood and timber and growing plants for raising goats. The objectives of this study were to compare the communities of filamentous fungi present in the preserved area and in the anthropized soil of the Catimbau National Park in Buíque, PE, Brazil, and to evaluate the impacts of anthropization on such communities. A total of 12 collections of soil samples were made, six in the preserved area and six in the anthropic area, and the physicochemical properties of the soil samples were analyzed. Fungi were isolated through suspension and serial dilution methods. After growth, the samples were purified and identified based on classical taxonomy, according to specific literature. The diversity, evenness, richness, dominance, frequency, and similarity among the species of filamentous fungi in both areas were assessed based on ecological indexes. A total of 4,488 colony-forming units of filamentous fungi were obtained, which were distributed into 65 species belonging to 15 genera. In the preserved area, higher abundance and richness of species were observed, with predominance of the genera Aspergillus and Penicillium. In both areas, diversity and equitability were high, demonstrating that the species are well distributed in these areas. In the preserved area, the dominant genera were Aspergillus, Gongronella, and Penicillium, whereas Aspergillus was the dominant genus in the anthropic area. Two distinct communities were observed in the areas analyzed. Principal component analysis showed that Penicillium simplicissimum influences the total diversity of both communities. The anthropization that occurred in the Catimbau National Park has changed the composition of the filamentous fungal communities of the site, restricting the number of species and decreasing the abundance of these important microorganisms. This results in ecosystem damage and likely causes relevant major imbalances, with serious consequences, such as possible disappearance of the aforementioned species, as well as of species yet undiscovered by the scientific community.
Tannase (tannin acyl hydrolse, E.C.3.1.1.20) is an enzyme that catalyzes the hydrolysis of ester and depside bonds of hydrolysable tannins. Among the filamentous fungi, the genus Penicillium is recognized as the second best producer of tannase. This enzyme presents applications in several industrial segments, such as in the production of judges and teas. The industrial production of tannase can be performed through solid state fermentation (SSF) by the use of tannin rich fruit residues, in order to minimize production costs. In the present study 31 species of Penicillium isolated from Caatinga and Atlantic Forest were tested quantitatively for the production of tannase. The species were kept in the Collection of Cultures Micoteca -URM (WDCM604) under mineral oil. The enzyme production was carried out under SSF using "cajá" (Spondias lutea L.) and Manga (Mangifera indica L.) residues that are rich in tannins. Mango residue proved to be the best inducer of the production of tannase by Penicillium species, most of which presented high tannase activity, ranging from 14.48 to 89.48 U mL . Therefore, P. rolfssi URM6216 is being indicated for optimization of tannase production by SSF, using mango residue as a substrate. This is the first study on the potential of hog-plum (cajá) and mango wastes as substrates for tannase production by filamentous fungi.
Tannase is an inducible extracellular enzyme produced by filamentous fungi, yeasts, and bacteria by solid-state fermentation (SSF) or submerged fermentation (SmF). Among the filamentous fungi, Aspergillus and Penicillium are recognized as the most efficient producers of tannase. The aims of this study were to evaluate the production of tannase under SSF by isolation from Aspergillus and Penicillium preserved in the Collection of Cultures Micoteca -URM (WDCM604); select the best enzyme producer, and use the crude extract to clarify mangaba and tamarind juices. The optimal conditions were determined by using the Placket-Burman Planning (PB) and response surface methodology (RSM). All tested crops produced activity between 2088.19 and 238.93 U/gds, and Aspergillus carneus URM5577 was the best producer. Through MSR, the best parameters for producing tannase were found to be 70 h of cultivation at pH 6.0, 7% tannic acid at 28°C and, as the response variable, 5449.31 activity U/gds. The optimum purification conditions were the molecular weight of PEG 8000 (g/mol), concentration of PEG 15% (w/w), 25% citrate (w/w), and pH 8.0. Its application in mangaba juice reduced the tannin content by 49.66% after 90 min and in tamarind by 51.82% at 120 min incubation at 37°C.
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