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

Self-fertility and polyembryony in South American yellow trumpet trees (Handroanthus chrysotrichus and H. ochraceus, Bignoniaceae): a histological study of postpollination events

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

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.

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.

Slaughterhouses fungal burden assessment: a contribution for the pursuit of a better assessment strategy

In slaughterhouses, the biological risk is present not only from the direct or indirect contact with animal matter, but also from the exposure to bioaerosols. Fungal contamination was already reported from the floors and walls of slaughterhouses. This study intends to assess fungal contamination by cultural and molecular methods in poultry, swine/bovine and large animal slaughterhouses. Air samples were collected through an impaction method, while surface samples were collected by the swabbing method and subjected to further macro- and micro-scopic observations. In addition, we collected air samples using the impinger method in order to perform real-time quantitative PCR (qPCR) amplification of genes from specific fungal species, namely A. flavus, A. fumigatus and A. ochraceus complexes. Poultry and swine/bovine slaughterhouses presented each two sampling sites that surpass the guideline of 150 CFU/m3. Scopulariopsis candida was the most frequently isolated (59.5%) in poultry slaughterhouse air; Cladosporium sp. (45.7%) in the swine/bovine slaughterhouse; and Penicillium sp. (80.8%) in the large animal slaughterhouse. Molecular tools successfully amplified DNA from the A. fumigatus complex in six sampling sites where the presence of this fungal species was not identified by conventional methods. This study besides suggesting the indicators that are representative of harmful fungal contamination, also indicates a strategy as a protocol to ensure a proper characterization of fungal occupational exposure.

Espécies de Aspergillus produtoras de aflatoxina, na região Araraquarense, SP

No presente trabalho foram isoladas, e estudadas quanto à produção de aflatoxina, espécies do gênero Aspergillus, principalmente as linhagens de A. flavus ocorrentes na região Araraquarense. o isolamento, seguindo métodos usuais em microbiologia, foram executados em amostras de amendoim, provenientes de duas épocas do ano, ou seja, da safra das «águas» e da safra da «seca». Após o isolamento as culturas foram classificadas. Dessa classificação pôde-se obter: 102 culturas de A. flavus; 4 culturas de A. oryzae; var. effusus; 2 culturas de A. oryzae; 2 culturas de A. parasiticus e 1 cultura do Grupo A. ochraceus. Durante os trabalhos 4 culturas foram perdidas, dessa forma foram estudadas realmente 107 culturas. Todas as culturas, a seguir, foram estudadas para a verificação de sua habilidade em produzir aflatoxina. A extração da toxina foi feita no micélio e no meio de cultura, usando-se técnicas padrões. A separação dos metabólitos foi feita em placas de camada delgada de silicagel, usando-se como solvente o benzeno-acetato de etila-etnol e quantificadas sob luz ultra-violeta. Analisando-se as 107 culturas de Aspergillus, notou-se que apenas 33 delas (31%) produziram aflatoxina, sendo que destas, 4 produriram apenas no meio de cultura, enquanto que duas produziram aflatoxina apenas no micélio. Observando-se ainda o comportamento das culturas que produziram aflatoxina, verificou-se que a produção da toxina no micélio foi muito maior que no meio da cultura, algumas produzindo elevadas quantidades (até 400 ppm de B1 e 300 ppm de G1). 3) Uma percentagem razoável de amostras (65% na série A e 70% na série B) produziram aflatoxina em pelo menos uma das colônias isoladas. 4) Algumas colônias produziram elevada quantidade de aflatoxina, chegando a 400 ppm de Bi e 300 ppm de d. 5) Uma baixa proporção do total das colônias (31%) produziu aflatoxina. 6) Dentre as demais espécies de Aspergillus apenas uma colônia de A. oryzae produziu aflatoxina e mesmo assim, só no meio de cultura. 7) Não houve correlação entre a produção de aflatoxina no meio de cultura e no micélio (r = 0,027), porém houve uma correlação fortemente positiva entre a produção de aflatoxina Bx e Gx no meio de cultura (r = 0,988) e também no micélio (r = 0,826).

The Adolpho Lutz collection of Tabanidae: (Diptera): I. The described genera and species, condition of the collection, and selection of lectotypes

A study of the Adolpho Lutz Collection of Tabanidae at the Instituto Oswaldo Cruz and of additional Lutz material at the Instituto Butantan in São Paulo is reported. Of the ninety-four species of Tabanidae validly described by Lutz, type material of eighty-four was recognized, either holotypes, allotypes or syntypes. Lectotypes were selected from among syntype series or remaining specimens and all type material was labelled. Of the ten species of which no type material could be found, neotypes were designated in the case of two species, Erephosis nigricans and Erephosis pseudo-aurimaculata. Types of three species, Chrysops ecuadoriensis, Dichelacera salvadorensis and Esenbeckia nigricorpus are believed to have been in Hamburg and destroyed during the last war. Types of two species, Esenbeckia biscutellata and E. dubia, and additional type material of several others are believed to have been in Montevideo. A request for information about them remains unanswered. Types of the remaining three species, Dichelacera intermedia, Dichelacera laceriascia and Esenbeckia distinguenda could not be found, and it is believed that at least the type of the last species was accidentally destroyed. Three specific of subspecific names proposed by Lutz but palaced by others in synonymy have been revalidated, Acanthocera intermedia, Erephosis brevistria and Esenbeckia fenestrata. Generic placement of two names has been changed, Esenbeckia arcuata ricardoae to Proboscoides, and Selasoma giganteum to Stibasoma. Seven specific names proposed by Lutz appear to be synonyms of earlier names, as follows: Bombylopsis juxtaleonina Lutz and Castro, 1936 = B. leonina Lutz, 1909. Bombylopsis pseudoanalis Lutz, 1909 = B. erythronotata (Bigot, 1892). Esenbeckia fuscipennis var. flavescens Lutz, 1909 = Esenbeckia fuscipennis Wied., 1828. Fidena chrysopyga Lutz and Castro, 1936 = F. atra Lutz and Castro, 1936. Laphriomyia longipalpis Lutz and Castro, 1937 = L. mirabilis Lutz, 1911. Stibasoma semiflavum Lutz, 1915 = St. bicolor Bigot, 1892. Tabanus hesperus Lutz, 1912 = Chlorotabanus (Cryptolylus) innotescens (Walker, 1854). Four Lutz names appear to antedate names proposed by others, viz.: Diachlorus angustifrons Kröber, 1930 and D. ochraceus Kröb., 1928 not Macquart, 1850 = Diachlorus fuscistigma Lutz, 1913. Psalidia fairchildi Barretto, 1950 = dicladocera conspicua Lutz and Neiva, 1914. Fidena pseudo-fulvithorax Kröb., 1931 = Erephopsis flavicrinis Lutz, 1909. Esenbeckia lemniscata Enderlein, 1925 = Esenbeckia clari Lutz, 1909. Some comments on Lutz' system of classification are given together with notes on the genotypes and included species of his genera as revaled by his collection and notes.

Identificação e características culturais de espécies do gênero Aspergillus isoladas de sementes de feijão no Estado da Paraíba

A partir de sementes de feijão (Phaseulus vulgaris) armazenadas no estado da Paraíba, foram isolados os fungos Aspergillus flavus, A. fumigatus, A. lucknowensis, A.niger, A.ochraceus e A. parasiticus. Os fungos foram identificados com base nas características culturais exibidas em meio de Czapek-Agar e morfológicas através de microscópio ótico composto. No oitavo dia de crescimento em meio de BDA, cada espécie apresentou diferenças quanto à curva de crescimento, taxa de crescimento durante o período de incubação e diâmetro das colônias no oitavo dia de acordo com a procedência das amostras de sementes.