Production and properties of xylanases from Aspergillus terricola Marchal and Aspergillus ochraceus and their use in cellulose pulp bleaching

Aspergillus terricola and Aspergillus ochraceus, isolated from Brazilian soil, were cultivated in Vogel and Adams media supplemented with 20 different carbon sources, at 30 A degrees C, under static conditions, for 120 and 144 h, respectively. High levels of cellulase-free xylanase were produced in birchwood or oat spelt xylan-media. Wheat bran was the most favorable agricultural residue for xylanase production. Maximum activity was obtained at 60 A degrees C and pH 6.5 for A. terricola, and 65 A degrees C and pH 5.0 for A. ochraceus. A. terricola xylanase was stable for 1 h at 60 A degrees C and retained 50% activity after 80 min, while A. ochraceus xylanase presented a t (50) of 10 min. The xylanases were stable in an alkali pH range. Biobleaching of 10 U/g dry cellulose pulp resulted in 14.3% delignification (A. terricola) and 36.4% (A. ochraceus). The brightness was 2.4-3.4% ISO higher than the control. Analysis in SEM showed defibrillation of the microfibrils. Arabinase traces and beta-xylosidase were detected which might act synergistically with xylanase.

Xylanases from Aspergillus niger, Aspergillus niveus and Aspergillus ochraceus produced under solid-state fermentation and their application in cellulose pulp bleaching

This study describes the production of xylanases from Aspergillus niveus, A. niger, and A. ochraceus under solid-state fermentation using agro-industrial residues as substrates. Enzyme production was improved using a mixture of wheat bran and yeast extract or peptone. When a mixture of corncob and wheat bran was used, xylanase production from A. niger and A. ochraceus increased by 18%. All cultures were incubated at 30 A degrees C at 70-80% relative humidity for 96 h. For biobleaching assays, 10 or 35 U of xylanase/g dry cellulose pulp were incubated at pH 5.5 for 1 or 2 h, at 55 A degrees C. The delignification efficiency was 20%, the brightness (percentage of ISO) increased two to three points and the viscosity was maintained confirming the absence of cellulolytic activity. These results indicated that the use of xylanases could help to reduce the amount of chlorine compounds used in cellulose pulp treatment.

Produção de quitosanase por aspergillus ochraceus em cultivo descontínuo submerso

In this work a 24 factorial design with triplicate at central point was used in order to investigate the influence of chitosan concentration (substrate) (Cs), culture media temperature (CMT), aeration ratio (AR) as well as agitation (A) on chitosanase production by Aspergillus ochraceus. Experiments were carried out using the following levels to the factors: (Cs) (-1) 0.1%; (0) 0.15%; (+1) 0.2%; (TMC) (-1) 25 minutes; (0) 30 minutes; (+1) 35 minutes; (RA) (-1) 0.4; (0) 0.6; (+1) 0.8; (A) (-1) 90 rpm, (0) 120 rpm, (+1) 150 rpm. One chitosanolytic activity (U.mL-1) was defined as the enzyme necessary to produce 1.0 mmol.min-1 of glicosamine by mL of extract. Chitosanolytic assays were carried out using two extract volumes, 0.05 and 0.1 mL, respectively. Results showed that was possible to produce chitosanase of order aproximatelly 5,9 U.mL-1 by Aspergillus ochraceus and chitosanolytic activity was increased by increment on substrate concentration, aeration ratio as well as agitation while media culture temperature increment decreased activity

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

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Control of amylase production and growth characteristics of Aspergillus ochraceus

The growth and the extracellular amylase production by Aspergillus ochraceus were studied in a stationary culture medium. Maximum growth rate of this fungus was found after 5 days of incubation at 30° C, but maximum amylase production was obtained after 2 days. The highest amylase production were attained with lactose, maltose, xylose and starch as carbon sources. The extracellular amylase production and mycelial growth were influenced by the concentration of starch. Other carbohydrates supported growth but did not induce amylase synthesis and glucose repressed it, indicating catabolite repression in this microorganism. The presence of both mechanisms of induction and repression suggests that at least these multiple forms of regulation are present in A. ochraceus. Of the nitrogen sources tested, casaminoacids, ammonium nitrate and sodium nitrate stimulated the highest yield of amylase. Optimal amylase production was obtained at pH 5.0, but enzyme activity was found only in the 4.0-6.0 pH range. These results were probably due to the inhibitory effect of NH 4 +-N in the culture medium.

Produção de biossurfactantes pelos fungos Aspergillus ochraceus e Penicillium expansum em fermentação semi-sólida utilizando resíduos agroindustriais como substrato

Pós-graduação em Microbiologia - IBILCE

Antifungal activity of flavonoids from Heteropterys byrsonimifolia and a commercial source against Aspergillus ochraceus: In silico interactions of these compounds with a protein kinase

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Xylanase and beta-Xylosidase Production by Aspergillus ochraceus: New Perspectives for the Application of Wheat Straw Autohydrolysis Liquor

The xylanase biosynthesis is induced by its substrate-xylan. The high xylan content in some wastes such as wheat residues (wheat bran and wheat straw) makes them accessible and cheap sources of inducers to be mainly applied in great volumes of fermentation, such as those of industrial bioreactors. Thus, in this work, the main proposal was incorporated in the nutrient medium wheat straw particles decomposed to soluble compounds (liquor) through treatment of lignocellulosic materials in autohydrolysis process, as a strategy to increase and undervalue xylanase production by Aspergillus ochraceus. The wheat straw autohydrolysis liquor produced in several conditions was used as a sole carbon source or with wheat bran. The best conditions for xylanase and beta-xylosidase production were observed when A. ochraceus was cultivated with 1% wheat bran added of 10% wheat straw liquor (produced after 15 min of hydrothermal treatment) as carbon source. This substrate was more favorable when compared with xylan, wheat bran, and wheat straw autohydrolysis liquor used separately. The application of this substrate mixture in a stirred tank bioreactor indicated the possibility of scaling up the process to commercial production.

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.

Identificação de Aspergillus spp: toxigênico em arroz

A composição química e o modo de cultivo do arroz o tornam susceptível à contaminação fúngica e, consequentemente, por micotoxinas. Considerando-se o expressivo consumo de arroz e a possibilidade de ser potencial fonte de micotoxinas, especial atenção deve ser dispensada quanto à qualidade do produto adquirido. Assim, o presente trabalho teve como objetivo avaliar espécies do gênero Aspergillus quanto à capacidade toxigênica, em diferentes subgrupos de arroz. As amostras constituíram-se de 31 marcas de arroz referentes aos subgrupos branco polido (21) e parboilizado (10), mais comumente comercializadas na cidade de Lavras - MG. Ao contrário dos outros subgrupos, a incidência de Aspergillus flavus e Aspergillus niger em amostras de arroz branco polido aumentou significativamente após a desinfecção. Pôde-se observar que, 50% dos Aspergillus flavus e 50% dos Aspergillus niger encontrados, foram considerados toxigênicos para o subgrupo branco polido. Na amostra de arroz parboilizado, 67% dos Aspergillus flavus eram potenciais produtores. O Aspergillus ochraceus não se revelou como toxigênico. Este estudo permitiu concluir que, apesar de trabalhos isolados, a presença de fungos toxigênicos em arroz é verídico, o que se torna relevante por se tratar de um cereal importante no cenário mundial.

Screening of Filamentous Fungi to Identify Biocatalysts for Lupeol Biotransformation

The goal of the study was to evaluate the ability of filamentous fungi to biotransform the pentacyclic triterpene lupeol. The microbial transformations were carried out in shake flasks in different media. Experiments were also run with control flasks. Samples of each culture were taken every 24 hours, extracted with ethyl acetate, and analyzed by GC-MS. The biotransformation of lupeol by Aspergillus ochraceus and Mucor rouxii afforded two compounds in each culture, which were detected in the cultures developed for more than seven days only in the Koch's K1 medium. The obtained data demonstrated that A. ochraceus is a good biocatalyst to introduce double bonds in the lupeol structure, whereas M. rouxii exhibits ability to biocatalyze oxygen insertions in that pentacyclic triterpene. Mass spectrometry was demonstrated to be an efficient analytical method to select promising biocatalysts for the compound investigated in this study. The biotransformation processes were influenced by the culture medium and incubation period. The obtained results open the perspective of using A. ochraceus and M. rouxii in pentacyclic triterpene biotransformations.

Fungos associados a grãos de café (Coffea arabica L.) beneficiados no sudoeste da Bahia

Diversos fatores podem interferir na qualidade do café, especialmente aqueles relacionados às etapas pós-colheita de processamento e secagem. Algumas espécies de fungos podem se associar a grãos de café durante a pós-colheita, podendo ocasionar alterações indesejáveis. O objetivo deste estudo foi avaliar a influência dos processamentos via seca (natural), seco em terreiro de terra, e via úmida (despolpado), seco em terreiro de cimento, tradicionalmente empregados na região sudoeste da Bahia, na incidência de fungos em grãos de café beneficiados produzidos na safra 2007/2008. O experimento consistiu de 4 tratamentos: a) café natural de Barra do Choça; b) café natural de Encruzilhada; c) café despolpado de Barra do Choça e d) café despolpado de Encruzilhada; e 5 repetições. Foram coletadas 20 amostras de grãos de café oriundas de diferentes propriedades cafeeiras nestes municípios. Os resultados obtidos foram avaliados pelo teste de médias t de Bonferroni a 5% de probabilidade. Houve diferença estatística significativa entre os tratamentos analisados para a infestação fúngica. Os gêneros detectados foram: Aspergillus, Penicillium e Fusarium, sendo que o gênero Aspergillus foi o de maior incidência, no qual foram identificadas oito espécies: Aspergillus ochraceus, A. niger, A. flavus, A. foetidus, A. tubingensis, A. auricomus, A. sojae e A. oryzae. Foi detectada a maior incidência de fungos em grãos de café oriundos de processamento natural do que de processamento despolpado.

Fungitoxicidade in vitro de iprodione sobre o crescimento micelial de fungos que se associam a sementes de arroz

Este estudo teve o objetivo de avaliar a fungitoxicidade de iprodione, em diferentes concentrações, sobre o crescimento micelial in vitro dos fungos mais comuns disseminados pelas sementes de arroz no Estado de Minas Gerais, Brasil. Isolados de Alternaria alternata, Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Curvularia oryzae, Drechslera oryzae, Gerlachia oryzae, Phoma sorghina e Pyricularia grisea foram obtidos de sementes de arroz e o seu cresci-mento micelial foi avaliado em meio BDA contendo o fungicida nas concentrações de 10mg.l-1, 50mg.l-1, 100mg.l-1 e 500mg.l-1, após sete dias de incubação. O efeito de iprodione foi avaliado por meio da ED50 (dose necessária para inibir em 50% o crescimento micelial). O fungicida iprodione inibe com alto efeito (ED50<10mg.l-1) o crescimento micelial dos fungos A. alternata, A. niger, A. ochraceus, C. oryzae, D. oryzae e G. oryzae, médio efeito (ED50 entre 10 e 50mg.l-1) Phoma sorghina e baixo efeito (ED50>50mg.l-1) Pyricularia grisea. Os fungos A. niger e A. ochraceus foram os mais sensíveis a iprodione, tendo apresentado ED50 de 2,5 e 3,1mg.l-1, respectivamente.

Avaliação do potencial de uso da restrição hídrica em teste de sanidade de sementes de trigo visando à detecção de fungos

Objetivou-se, com o presente trabalho, avaliar a viabilidade do uso da restrição hídrica, em substituição ou alternativa ao uso da técnica de congelamento de sementes de trigo, no teste de incubação em substrato de papel, blotter test. O trabalho consistiu na comparação das duas técnicas por meio do exame de 60 amostras de sementes, coletadas em diferentes áreas do Estado de Minas Gerais e outras regiões do país, tendo como restritor hídrico o soluto manitol, no potencial de -1,10 MPa, valor este determinado preliminarmente com base na capacidade de redução da germinação das sementes de trigo em condições do teste de sanidade. A incidência média de Bipolaris sorokiniana não foi afetada por nenhuma das técnicas comparadas, havendo, por outro lado, um maior estímulo da restrição hídrica em relação à Pyricularia grisea, Aspergillus ochraceus, Cladosporium cladosporioides e Epicoccum purpuracens. Para Alternaria alternata, Drechslera tritici-repentis e Fusarium graminearum, os maiores valores de incidência ocorreram no método do congelamento.

Identification of the antimicrobial substances produced by Solanum palinacanthum (Solanaceae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)