Behavior of the Brown-rot Fungus Gloeophyllum trabeum on Thermally-modified Eucalyptus grandis Wood

ABSTRACT In this study, we aimed evaluate the behavior of the brown-rot fungus Gloeophylum trabeum and white-rot fungus Pycnoporus sanguineus on thermally-modified Eucalyptus grandis wood. To this end, boards from five-year-eleven-month-old E. grandis trees, taken from the Duratex-SA company stock, were thermally-modified between 180 °C and 220 °C in the Laboratory of Wood Drying and Preservation at Universidade Estadual Paulista -UNESP, Botucatu, Sao Paulo state Brazil. Samples of each treatment were tested according to the ASTM D-2017 (2008) technical norm. The accelerated decay caused by the brown-rot fungus G. trabeum was compared with the decay caused by the white-rot fungus P. sanguineus, studied b

Modification of lignin structure caused by fungi

O licor negro, cuja composição majoritária é de lignina, é o principal subproduto gerado após a obtenção de polpa celulósica. Este polímero natural é caracterizado pela elevada massa molecular (MM) e composição aromática. Processos físico-químicos são empregados na fragmentação da lignina com o objetivo de obter compostos aromáticos, cuja fonte não renovável é oriunda de derivados do petróleo. A biodegradação da lignina, causada pela ação de enzimas ligninolíticas fúngicas pode prover compostos aromáticos de baixo peso molecular, como fenóis e antioxidantes, utilizados em diversos segmentos industriais. O objetivo deste trabalho foi estudar a modificação estrutural da lignina causada por fungos basidiomicetos coletados de mata nativa e campos de reflorestamento de eucalipto. Os ensaios de biodegradação foram inicialmente conduzidos utilizando-se 10 espécies de fungos, cultivados em duas concentrações de licor negro (10% e 15%, v/v). No ensaio com licor menos concentrado, utilizaram-se 10 espécies, e no ensaio com licor 15% utilizou-se somente a espécie Pycnoporus sanguineus. As análises realizadas para o estudo da modificação estrutural da lignina foram: espectroscopia na região do infravermelho (FTIR/ATR), cromatografia líquida de gel filtração e espalhamento de raios-X a baixo ângulo (SAXS). P. sanguineus e espécies de fungos do gênero Trametes possuem elevada capacidade de consumir a lignina presente no licor 10%, após 14 dias. A atividade de enzimas ligninolíticas de P. sanguineus foi investigada nas duas concentrações de licor. No licor 10%, observou-se elevada atividade das enzimas ligninolíticas manganês peroxidase (MnP), lacase e peroxidases; somente a MnP foi detectada com maior atividade no licor 15%. Análises de Demanda Bioquímica de Oxigênio (DBO) indicaram que o licor negro 10% biodegradado por P. sanguineus possui elevada carga de DBO, sendo tóxico a Daphia magna e Hydra attenuatta. Análises de metabólitos produzidos pelos fungos no licor negro 10% por cromatografia gasosa acoplada à espectrometria de massas (GC-MS) indicaram, principalmente, a produção de açúcares e álcoois em concentrações não muito diferentes dos seus respectivos controles. Embora os fungos sejam capazes de reduzir a massa molecular da lignina presente no licor negro, compostos derivados da sua fragmentação podem sofrer reações de condensação ou repolimerização, contribuindo, eventualmente, para elevar sua massa molecularBlack liquor which the bulk content is lignin, is the main product generated after obtaining cellulose pulp. This natural polymer is characterized by high molecular weight (MW) and aromatic compounds. Physical-chemical processes can be employed to fragment lignin and to produce aromatic compounds to replace those from non-renewable sources such as petroleum. Biodegradation of lignin can provide low molecular weight compounds, such as phenols and antioxidants, which may be employed in various industrial processes. The aim of this work was to investigate the structural modification of lignin caused by basidiomycetes fungi, obtained from native forest and Eucalyptus fields. The biodegradation assay was done with10 fungi species cultivated with two concentrations of black licor (10% and 15%, v/v). The assay with 10% liquor, 10 species of basidiomycete fungi were used, and in the assay with 15% liquor, it was used only Pycnoporus sanguineus. The analyses carried out to determine the structural modification of lignin were: infrared spectroscopy (FTIR/ATR), liquid chromatography gel filtration and small angle X-ray scattering (SAXS). P. sanguineus and fungi of Trametes genera had high capacity to consume the lignin content in the 10% liquor, after 14 days of cultivation. After that, P. sanguineus was selected for assays of ligninolytic enzymes activities in the two liquor concentrations. High activities of manganese peroxidase (MnP), laccase and peroxidase were detected in the 10% liquor, whereas in the concentration of 15% only high MnP activity was detected. Biochemical Oxygen Demand, analysis (BOD) indicated that the biodegraded liquor had high BOD load, and it was toxic to Daphia magna and Hydra attenuata. Metabolites produced by the fungi in the liquor 10% were analysed by gas chromatography-mass spectrometry (GC-MS), which indicated mainly the production of sugars and alcohols in concentrations not far from their respective controls. Although the fungus is able to decrease molecular weight of lignin, its fragments derivatives can undergo condensation or repolymerization reactions, contributing, eventually, to increase its molecular weigh

Seleção de fungos degradadores de madeira para uso na destoca biológica de Eucalyptus spp

O eucalipto é a arbórea mais plantada no Brasil, devido ao seu amplo uso e após seu corte, os tocos e raízes que permanecem no local, dificultam o manejo da cultura. Com base nessa necessidade, o objetivo do presente estudo foi avaliar as características fisiológicas e funcionais de fungos basidiomicotas, isolados de campos de reflorestamento de Eucalyptus spp., com vistas à possibilidade do uso destes na destoca natural. Os fungos Pycnoporus sanguineus, Lentinus bertieri, Xylaria sp. e Lentinula edodes e um isolado não identificado foram estudados. No primeiro experimento, avaliou-se a patogenicidade dos mesmos em plantas de eucalipto urograndis (10 meses de idade), através da inoculação de disco de meio de cultura colonizado e não colonizado (Testemunha). As avaliações foram feitas aos 30 e 60 dias após a inoculação, com base no tamanho das lesões internas e externas. No segundo experimento, avaliou-se o crescimento micelial dos fungos P. sanguineus, L. bertieri, Xylaria sp., L. edodes e S. ostrea, em substrato à base de serragem enriquecida com farelos, e em três composições de meios de cultura, Batata-Dextrose-Ágar (BDA); Malte-Ágar (MA) e Serragem-Dextrose-Ágar (SDA), e incubados a 23, 27 e 31ºC. No terceiro experimento, avaliou-se a produção das enzimas ligninocelulolíticas lacase e celulase, e outras ligadas à degradação de tecidos vegetais, as proteases, pectinases e lipase, em meios de cultura específicos. No quarto experimento, estudou-se, pelo método “soil block”, a perda de massa (%) de corpos-de-prova de eucalipto urograndis, inoculados com os fungos P. sanguineus, L. bertieri, Xylaria sp. e L. edodes, mantidos sob dois regimes de umidade (50 e 100%), e avaliados aos 30, 60, 90 e 120 após inoculação. Estudaram-se também as características químicas da madeira, após 120 dias da inoculaçãoThe Eucalyptus spp. tree is the most planted in Brazil, due to its widespread use and after its use, the stumps and roots that remains in the area, hinder crop management. Based on this need, the purpose of this work was evaluate the physiological and functional characteristics of basidiomycetes fungi, obtained in reforestation fields of Eucalyptus spp., with the possibility of using the fungi in natural stump removal. Pycnoporus sanguineus, Lentinus bertieri, Xylaria sp., Stereum ostrea and a unidentified isolate, were studied. In the first, was evaluate the fungi pathogenicity in eucalipt urograndis plants (10 months old), by means of disk inoculation into culture medium colonized and non-colonized (control) by fungi. Evaluations were performed at 30 and 60 days after inoculation based on the external and internal size of lesions. In experiment 2, the mycelial growth of P. sanguineus, L. bertieri, Xylaria sp., L. edodes and S. ostrea was evaluated in bran and sawdust-based substrate, as well as in three culture media: Potato-Dextrose-Agar (PDA), Malt-Agar (MA) and Sawdust-Dextrose-Agar (SDA), and at three incubation temperatures, at 23, 27 and 31ºC. In experiment 3, the production of lignolytic enzymes lacase and cellulase, and other enzymes related to degradation of plant tissues, like protease, pectinases and lipase was assessed in specific culture media. In experiment 4, mass loss (%) and chemical features of eucalipt urograndis wood degraded by the soil block method were evaluated by the P. sanguineus, L. bertieri, Xylaria sp. and L. edodes fungi. Thus, test specimens were inoculated with the fungi and kept under two moisture regimes, 50 and 100%, and incubated at 25 ±2ºC. Mass loss was assessed at 30, 60, 90 and 120 days after inoculation. The chemical features of eucalipt urograndis test specimens were only analyzed at 120 daysCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Deterioration of Wood Plastics Composites by the White-Rot Fungus Pycnoporus sanguineus

Wood plastic composites (WPC) are characterized by the mixing of wood fibers with plastics, allowing the production of new products whose characteristics are in several aspects superior to those of the original products and represent an expanding class of durable and low-cost materials in which their uses can reduce the environmental footprint and the dependence on petroleum products. Nevertheless, WPC has some setbacks, including biodegradation, which shortens its life span. In this study, the wood composite was exposed to the white-rot fungus Pycnoporus sanguineus in order to evaluate its resistance to biodegradation. The WPC was prepared with a 1:1 ratio of Eucalyptus spp. bark as reinforcement agent and polypropylene as matrix. Mechanical and rheological properties and mass loss were evaluated from 15 to 120 days of fungus exposure. After 15 days, a mass loss was detected, which transmitted a negligible effect on the impact resistance of the composite. For the 120-day fungus-exposed composite, the fungus produced a biofilm under the WPC that create a special environment for lignocellulosic consuming led to deterioration of the mechanical properties and minor changes on the thermal–chemical stability of the WPC. Finally, the study gave a great indication of the susceptibility of a Eucalyptus-based composite to biodegradation

Fragmentation of Lignin from Organosolv Black Liquor by White Rot Fungi

The mycelial growth ability of 13 white rot fungi were separately evaluated in kraft, organosolv, and soda black liquor agar-plates. The fungus able to best grow and decolorize black liquor agar-plates was grown in organosolv black liquor to investigate whether it reduced organosolv lignin molar mass. The fungus Bjerkandera adusta showed fair mycelial growth and decolorization ability in 10% black liquor-agar plates. To obtain low-molecular weight (MW) lignin, B. adusta was cultivated in 150-mL Erlenmeyer flasks containing 10% black liquor and maintained in a shaking culture for 15 days. Lignin was recovered from each Erlenmeyer flask by acid precipitation and was analysed by size exclusion chromatography (SEC) and Fourier transform infrared (FTIR) spectroscopy. The lowest MW of lignin from black liquor was observed on the 11th and 12th days, at 1461 and 1790 kDa, respectively, with the polydispersity close to 1.0, indicating that the molecules were similar in size. Fourier transform infrared spectra bands showed modification of the lignin structure during 9 days, with new bands appearing after five days of lignin biodegradation

Fragmentation of Lignin from Organosolv Black Liquor by White Rot Fungi

The mycelial growth ability of 13 white rot fungi were separately evaluated in kraft, organosolv, and soda black liquor agar-plates. The fungus able to best grow and decolorize black liquor agar-plates was grown in organosolv black liquor to investigate whether it reduced organosolv lignin molar mass. The fungus Bjerkandera adusta showed fair mycelial growth and decolorization ability in 10% black liquor-agar plates. To obtain low-molecular weight (MW) lignin, B. adusta was cultivated in 150-mL Erlenmeyer flasks containing 10% black liquor and maintained in a shaking culture for 15 days. Lignin was recovered from each Erlenmeyer flask by acid precipitation and was analysed by size exclusion chromatography (SEC) and Fourier transform infrared (FTIR) spectroscopy. The lowest MW of lignin from black liquor was observed on the 11th and 12th days, at 1461 and 1790 kDa, respectively, with the polydispersity close to 1.0, indicating that the molecules were similar in size. Fourier transform infrared spectra bands showed modification of the lignin structure during 9 days, with new bands appearing after five days of lignin biodegradation

Potential application of Ganoderma lucidum in solid state fermentation of primary sludge and wheat straw

This study was conducted to investigate the production of lignocellulolytic enzymes and sugars by the fungus Ganoderma lucidum during solid state fermentation (SSF) using primary sludge (PS) and wheat straw (WS) as substrates at different concentration ratios. For fungal growth on SSF, 20 g of each blended substrate was added to Erlenmeyer flasks, which were autoclaved and maintained at room temperature prior to inoculation, whereas for submerged fermentation (SF), flasks containing 25 mL of potato dextrose broth (PDB) were used as standard to check the differences between both methods of growth, and then all flasks were incubated at 25 degrees C in the dark, during 8 and 16 days. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis from the protein extract obtained from solid state fermentation strongly suggested that G. lucidum could produce lignocellulolytic enzymes to degrade primary sludge and wheat straw. Among the sugars, the production of xylose and mannose was disturbed by adding primary sludge. With the addition of primary sludge, high glucuronic acid content was observed. The results suggest that the combination of primary sludge and wheat straw, at concentration ratios of 1: 1 to 1: 3, respectively, can be used as a raw material in the production of lignocellulolytic enzymes and the bioconversion of other types of biomass by G. lucidum.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Behavior of the brown-rot fungus Gloeophyllum trabeum on thermally-modified Eucalyptus grandis wood

In this study, we aimed evaluate the behavior of the brown-rot fungus Gloeophylum trabeum and white-rot fungus Pycnoporus sanguineus on thermally-modified Eucalyptus grandis wood. To this end, boards from five-year-eleven-month-old E. grandis trees, taken from the Duratex-SA company stock, were thermally-modified between 180 ºC and 220 ºC in the Laboratory of Wood Drying and Preservation at Universidade Estadual Paulista - UNESP, Botucatu, Sao Paulo state Brazil. Samples of each treatment were tested according to the ASTM D-2017 (2008) technical norm. The accelerated decay caused by the brown-rot fungus G. trabeum was compared with the decay caused by the white-rot fungus P. sanguineus, studied by Calonego et al. (2010). The results showed that (1) brown-rot fungus caused greater decay than white-rot fungus; and (2) the increase in temperature from 180 to 220 ºC caused reductions between 28.2% and 70.0% in the weight loss of E. grandis samples incubated with G. trabeum.O objetivo deste estudo foi avaliar o comportamento do fungo de podridão parda Gloeophylum trabeum e do fungo de podridão branca Pycnoporus sanguineus sobre a madeira de Eucalyptus grandis modificada termicamente. Tábuas de árvores de E. grandis com cinco anos e 11 meses de idade, da empresa Duratex-SA, foram modificadas termicamente entre 180 ºC e 220 ºC no Laboratório de Secagem e Preservação de Madeiras da UNESP, Botucatu-SP, Brasil. Corpos de prova de cada tratamento foram testados, de acordo com a norma técnica ASTM D-2017 (2008). O apodrecimento acelerado causado pelo fungo de podridão parda G. trabeum foi comparado com o do fungo de podridão branca P. sanguineus, estudado por Calonego et al. (2010). Os resultados mostraram que (1) o apodrecimento causado pelo fungo de podridão parda foi maior que o de podridão branca e (2) o aumento da temperature de 180 para 220 ºC ocasionou reduções de 28,2% a 70,0% na perda de massa dos corpos de prova de E. grandis incubados com o G. trabeum

Behavior of the brown-rot fungus Gloeophyllum trabeum on thermally-modified Eucalyptus grandis wood

In this study, we aimed evaluate the behavior of the brown-rot fungus Gloeophylum trabeum and white-rot fungus Pycnoporus sanguineus on thermally-modified Eucalyptus grandis wood. To this end, boards from five-year-eleven-month-old E. grandis trees, taken from the Duratex-SA company stock, were thermally-modified between 180 ºC and 220 ºC in the Laboratory of Wood Drying and Preservation at Universidade Estadual Paulista - UNESP, Botucatu, Sao Paulo state Brazil. Samples of each treatment were tested according to the ASTM D-2017 (2008) technical norm. The accelerated decay caused by the brown-rot fungus G. trabeum was compared with the decay caused by the white-rot fungus P. sanguineus, studied by Calonego et al. (2010). The results showed that (1) brown-rot fungus caused greater decay than white-rot fungus; and (2) the increase in temperature from 180 to 220 ºC caused reductions between 28.2% and 70.0% in the weight loss of E. grandis samples incubated with G. trabeum