Characterization of a thermostable extracellular tannase produced under submerged fermentation by Aspergillus ochraceus

Background: Tannases are enzymes that may be used in different industrial sectors as, for example, food and pharmaceutical. They are obtained mainly from microorganisms, as filamentous fungi. However, the diversity of fungi stays poorly explored for tannase production. In this article, Aspergillus ochraceus is presented as a new source of tannase with interesting features for biotechnological applications. Results: Extracellular tannase production was induced when the fungus was cultured in Khanna medium with tannic acid as carbon source. The extracellular tannase was purified 9-fold with 2% recovery and a single band corresponding to 85 kDa was observed in SDS-PAGE. The native apparent molecular mass was estimated as 112 kDa. Optima of temperature and pH were 40 degrees C and 5.0, respectively. The enzyme was fully stable from 40 degrees C to 60 degrees C during 1 hr. The activity was enhanced by Mn2+ (33-39%) and NH4+ (15%). The purified tannase hydrolyzed tannic acid and methyl gallate with Km of 0.76 mM and 0.72 mM, respectively, and Vmax of 0.92 U/mg protein and 0.68 U/mg protein, respectively. The analysis of a partial sequence of the tannase encoding gene showed an open read frame of 567 bp and a sequence of 199 amino acids were predicted. TLC analysis revealed the presence of gallic acid as a tannic acid hydrolysis product. Conclusion: The extracellular tannase produced by A. ochraceus showed distinctive characteristics such as monomeric structure and activation by Mn2+, suggesting a new kind of fungal tannases with biotechnological potential. Further, it was the first time that a partial gene sequence for A. ochraceus tannase was described.

Hofmeister effects on the colloidal stability of poly(ethylene glycol)-decorated nanoparticles

The colloidal stability of poly(ethylene glycol)-decorated poly(methyl methacrylate), PMMA/Tween-20, particles was investigated by means of phase separation measurements, in the presence of sodium fluoride (NaF), sodium chloride, sodium bromide, sodium nitrate, or sodium thiocyanate (NaSCN) at 1.0 mol L-1. Following Hofmeister's series, the dispersions of PMMA/Tween-20 destabilized faster in the presence of NaF than with NaSCN. After the phase separation, the systems were homogenized and except for the dispersions in NaF, re-dispersed particles took longer to destabilize, indicating that anions adsorbed on the particles, creating a new surface. Except for F- ions, the adsorption of anions on the polar outmost shell was evidenced by means of tensiometry and small-angle X-ray scattering measurements. Fluoride ions induced the dehydration of the polar shell, without affecting the polar shell electron density, and the formation of very large aggregates. A model was proposed to explain the colloidal behavior in the presence of Hofmeister ions.

Sintering and Microwave Properties of Zirconium Tin Titanate Doped with Select Oxides

Zirconium tin titanate (ZST) is often used as a dielectric resonator for the fabrication of microwave devices. Pure compositions do not sinter easily by solid state sintering; therefore, sintering ZST requires sintering aids capable of creating defects that could improve diffusion processes and/or promote liquid phase sintering. The mechanisms by which the additives influence the microstructure and, consequently, the ZSTs dielectric properties are not very clear. The effects of ZnO, Bi2O3, and La2O3, on the stoichiometry and dielectric properties of ZST sintered at different temperatures were investigated in this study.

Bioprocess and biotecnology: effect of xylanase from Aspergillus niger and Aspergillus flavus on pulp biobleaching and enzyme production using agroindustrial residues as substract

This study compares two xylanases produced by filamentous fungi such as A. niger and A. flavus using agroindustrial residues as substract and evaluated the effect of these enzymes on cellulose pulp biobleaching process. Wheat bran was the best carbon source for xylanase production by A. niger and A. flavus. The production of xylanase was 18 and 21% higher on wheat bran when we compare the xylanase production with xylan. At 50°C, the xylanase of A. niger retained over 85% activity with 2 h of incubation, and A. flavus had a half-life of more than 75 minutes. At 55°C, the xylanase produced by A. niger showed more stable than from A. flavus showing a half-life of more than 45 minutes. The xylanase activity of A. niger and A. flavus were somehow protected in the presence of glycerol 5% when compared to the control (without additives). On the biobleaching assay it was observed that the xylanase from A. flavus was more effective in comparison to A. niger. The kappa efficiency corresponded to 36.32 and 25.93, respectively. That is important to emphasize that the cellulase activity was either analyzed and significant levels were not detected, which explain why the viscosity was not significantly modified.

Uso de aditivos na biodegradação de madeira pelo fungo Ceriporiopsis subvermispora: efeito na peroxidação de lipídios dependente de manganês-peroxidase

Ceriporiopsis subvermispora is a selective fungus in the wood delignification and the most promising in biopulping. Through the lipid peroxidation initiated by manganese peroxidase (MnP), free radicals can be generated, which can act in the degradation of lignin nonphenolic structures. This work evaluated the prooxidant activity (based in lipid peroxidation) of enzymatic extracts from wood biodegradation by this fungus in cultures containing exogenous calcium, oxalic acid or soybean oil. It was observed that MnP significant activity is required to promote lipid peroxidation and wood delignification. Positive correlation between prooxidant activity x MnP was observed up to 300 IU kg-1 of wood.

Adição de carotenóides naturais e artificiais na alimentação de galinhas poedeiras: efeitos na qualidade de ovos frescos e armazenados

A pesquisa teve como objetivo avaliar os atributos de qualidade e de durabilidade dos ovos de duas diferentes linhagens de poedeiras, com dietas contendo aditivos artificiais ou naturais, em diferentes períodos de armazenamento. Foram quantificadas durante o armazenamento as variáveis perda de peso, pH da gema e albúmen, gravidade específica, índice de gema (IG) e foi realizada também classificação por peso e Unidade Haugh (UH). Os resultados foram avaliados segundo teste de Tukey (P≤0,05). As linhagens diferiram quanto à classificação extra dos ovos, com maior porcentagem para a Isa Brown. Os ovos obtidos das poedeiras Carijó apresentaram maior porcentagem de gema (Carijó 30,66 e Isa-Brown 24,55). O tempo de armazenamento fez com que a porcentagem de gema aumentasse nas duas linhagens (Carijó 30,66 tempo zero para 34,85 depois de 36 dias; e Isa- Brown de 24,55 para 27,48). As dietas com aditivos artificiais ou naturais não influenciaram a porcentagem de clara (% Clara 58,72 Controle; 59,52 Aditivo; 60,11 Urucum 1,5%; 59,43 e de gema %Gema 27,64 Controle; 27,90 Aditivo; 27,21 Urucum 1,5%; 27,66 Urucum 2%). O pH da clara diferiu entre os tratamentos no tempo inicial e aos 36 dias de armazenamento, mas o pH da gema não diferiu durante todo o armazenamento. O índice de gema diferiu nos tempos de armazenamento, mas não diferiu considerando os tratamentos. Em dietas com aditivos artificiais ou naturais, as linhagens e os tempos de armazenamento influenciam na qualidade e durabilidade dos ovos.

Effects of microbial inoculants and amino acid production by-product on fermentation and chemical composition of sugarcane silages

The objective of this study was to evaluate the chemical composition, fermentation patterns and aerobic stability of sugarcane silages with addition of amino acid production (monosodium glutamate) by-product (APB) and microbial inoculants. Mature sugarcane was chopped and ensiled in laboratory silos (n = 4/treatment) without additives (control) and with APB (10 g/kg), Pioneer 1174® (PIO, 1.0 mg/kg, Lactobacillus plantarum + Streptoccoccus faecium, Pioneer), Lalsil Cana (2.0 mg/kg, Lactobacillus buchineri, Lallemand) or Mercosil Maís 11C33® (1.0 mg/kg, Lactobacillus buchineri + Lactobacillus plantarum + Streptoccoccus faecium, Timac Agro). Fresh silage and silage liquor samples were obtained to assess pH, chemical composition and organic acid concentrations. Silage temperature was recorded throughout seven days to evaluate aerobic stability. The addition of APB decreased lactic acid levels, increased pH and N-NH3 and did not alter ethanol, acetic and butyric acids concentrations or dry matter (DM) losses. Microbial inoculants enhanced acetic acid levels, although only Pioneer 1174® and Mercosil Maís 11C33® lowered ethanol, butyric acid and DM losses. The addition of APB increased CP content and did not modify DM, soluble carbohydrates contents or in vitro dry matter digestibility. Additives did not alter silage maximum temperature or temperature increasing rate; however, Pioneer 1174® and Mercosil Maís 11C33® increased the time elapsed to reach maximum temperature. Monosodium glutamate production by-product does not alter fermentation patterns or aerobic stability of sugarcane silages, whereas homofermentative bacteria can provide silages of good quality.

Contribuição da inclusão de polpa cítrica e casca de soja para a qualidade da silagem de resíduo úmido de cervejaria

Com o objetivo de avaliar a produção da silagem e o uso de aditivos no processo de ensilagem do resíduo úmido de cervejaria, foram realizados 5 tratamentos: controle (C: ensilagem de 100% de resíduo úmido de cervejaria); PC15 (15% de polpa cítrica); PC30 (30% de polpa cítrica); CS15 (15% de casca de soja); CS30 (30% de casca de soja) – com base na matéria fresca do resíduo de cervejaria. As silagens foram confeccionadas em baldes plásticos com 252mm de altura e 245mm de diâmetro (0,06174m³), e amostras foram coletadas para análises bromatológicas, pH, nitrogênio amoniacal, digestão in vitro de matéria seca, ácidos orgânicos e perfil microbiológico. Os resultados foram analisados pelo programa computacional Statistical Analysis System (Statistical..., 1985), sendo verificada a normalidade dos resíduos pelo Teste de Shapiro-Wilk (PROC UNIVARIATE), e as variâncias, pelo Teste de Hartley. Os efeitos dos níveis de adição foram separados por meio de contrastes polinomiais utilizando o nível de significância de 5%. Houve aumento do teor de matéria seca, carboidratos solúveis, ácido lático, digestão in vitro de matéria seca, da população de bactérias ácido láticas e redução do pH, ácido butírico, propiônico e nitrogênio amoniacal a partir das inclusões de polpa cítrica e casca de soja, sendo os melhores resultados encontrados para o tratamento com inclusão de 30% de polpa cítrica (P<0,05). A ensilagem do bagaço de malte por si só é uma alternativa para o produtor rural como suporte alimentar e confecção de silagem de qualidade que pode ser incrementada com o uso de aditivos a serem avaliados de acordo com a relação custo-benefício para eficiência da produção