Cellulolytic activity of anaerobic facultative fungi from the digestive tract of sheep fed with banana leaf hay / Atividade celulolítica de fungos aeróbios facultativos do trato digestório de ovinos alimentados com feno da folha de bananeira

The addition of cellulolytic fungi, or their enzymes, in diets containing high levels of fiber are promising strategy for improving the performance. In this study, the aims were to select cellulolytic fungi from the digestive tract of sheep fed different concentrations of banana leaf hay. Thirty lambs raised in a feedlot were evaluated, distributed in a completely randomized design, with diets containing 0, 125, 250, 375, or 500 g/Kg of dry matter and six replications. Approximately 15 mL of ruminal fluid and swabs from the rectal ampulla were collected. The cultures were carried out in a medium containing microcrystalline cellulose (C medium). The mycelial fungi isolates were identified through the microculture technique. Among the fungi from the ruminal fluid, 23 isolates corresponded to the genus Aspergillus and three to Paecilomyces spp. Among the isolates from the rectal ampulla, seven were A. spp., and three were P. spp. The A. genus predominated among the isolates from both evaluated sites (p <0.05). Fragments of these fungi were inoculated in triplicate in medium C at 37 °C and the cellulolytic activity index (CAI) was determined after 24, 48, and 72 hours of incubation. There was no difference in the CAI of Aspergillus spp. from animals fed different diets or of different evaluated sites (P > 0.05). However, 22 isolates of Aspergillus spp. and three of Paecilomyces spp. showed a CAI > 1, indicating biotechnological potential for cellulase production. These selected isolates could be selected for the elaboration of microbial additives in ruminant diets.(AU)

A adição de fungos celulolíticos, ou suas enzimas, em dietas contendo elevados teores de fibras são estratégias pro-missoras para melhorar o desempenho. Neste estudo os objetivos foram selecionar fungos celulolíticos do trato digestório de ovinos alimentados com diferentes concentrações do feno da folha da bananeira (FBH). Foram avaliados 30 borregos criados em sistema intensivo, distribuídos em delineamento inteiramente ao acaso, com cinco dietas contendo 0, 125, 250, 375 ou 500 g/KG de matéria seca em seis repetições. Foram coletados aproximadamente 15 mL de fluido ruminal e swabs da ampola retal. Os cultivos foram realizados em meio de cultura contendo celulose microcristalina (meio C). Os fungos micelianos foram identificados após a técnica de microcultivo. Entre os fungos provenientes do fluido ruminal, 23 isolados corresponderam ao gênero Aspergillus e três a Paecilomyces spp.. Foram identificados nas fezes dos animais sete Aspergillus spp. e três Paecilomyces spp.. O gênero Aspergillus predominou entre os isolados de ambos os sítios avaliados (p =0,013). Fragmentos desses fungos foram inoculados em triplicada em meio C a 37 °C e determinou-se o índice de atividade celulolítica(IAC) após 24, 48 e 72 horas de incubação. Não houve diferença entre CAI de isolados de Aspergillus spp. provenientes dos animais em diferentes dietas ou sítios avaliados (P > 0.05). Entretanto, 22 isolados de Aspergillus spp. e três de Paecilomyces spp. apresentaram IAC >1, indicando potencial biotecnológico para produção de celulases.(AU)

Fungal cellulases: production by solid-state cultivation in packedbed bioreactor using solid agro-industrial by-products as substrates and application for hydrolysis of sugarcane bagasse / Celulases fúngicas: produção por cultivo sólido em biorreator de leito empacotado usando resíduos sólidos agroindustriais como substratos e aplicação na hidrólise de bagaço de cana

Cellulases are essential for the hydrolysis of lignocellulosic materials in the production of second generation ethanol. Solid-state cultivation is a process that provides high concentrations of enzymes that can be used in this hydrolysis. The objectives of this work were to produce cellulases by cultivating the fungus Myceliophthora thermophila I-1D3b in a packed bed bioreactor with sugarcane bagasse (SCB) and wheat bran (WB) as substrate and to evaluate the efficiency of the enzymatic extract in the hydrolysis of SCB in natura (BIN) and pretreated with ozone, alkali and ultrasound (BOU). The conditions for enzyme production in the bioreactor were SCB:WB at a ratio of 2.3:1 (w/w), 75 % moisture content; 45 ºC; aeration rate 240 L h-¹ and 96 h. The enzyme production was evaluated by endoglucanase, xylanase, filter paper (FPU) and β-glycosidase activities. For the application of the enzymes, a central composed response surface design with 5 repetitions of the central point was used, taking enzyme volume and hydrolysis time as factors. Such cultivation yielded the following enzymatic activities: 723 U gss-¹ of endoglucanases, 2024 U gss-¹ of xylanase, 12.6 U gss-¹ of FPU and 41 U gss-¹ of β-glucosidase. The results of the application tests indicated the best conditions as 7.0 mL of the enzyme extract (4.2 FPU) and 6 hours for BIN and BOU. The best cellulose-glucose conversions were obtained for BOU, reaching 32.1 % at 65 ºC. In conclusion, the enzyme production in the packed bed bioreactor was efficient and BOU pretreatment improved the hydrolysis of biomass, increasing the efficiency of conversion of cellulose to glucose.

Celulases são essenciais para a hidrólise de materiais lignocelulósicos visando à produção de etanol de segunda geração. O cultivo em estado sólido é um processo que proporciona altas concentrações de enzimas que podem ser aplicadas nessa hidrólise. O objetivo deste trabalho foi produzir celulases pelo fungo Myceliophthora thermophila I-1D3b em biorreator de leito empacotado com bagaço decana (BC) e farelo de trigo (FT) como substrato e avaliar a eficiência deste extrato enzimático na hidrólise de bagaço de cana in natura (BIN) e pré-tratado com ozônio, álcali e ultrassom (BOU). As condições para produção de enzimas no biorreator foram BC:FT numa proporção de 2,3:1 (m/m); 75 %de umidade em base úmida; 45 ºC; taxa de aeração 240 L h-1 e 96 h. A produção de enzimas foi avaliada pelas atividades de endoglucanase, xilanase, papel de filtro (FPU) e β-glicosidase. Para a aplicação das enzimas, foi utilizado um planejamento de superfície de resposta central composto com 5 repetições do ponto central, tendo como fatores o volume de extrato enzimático e o tempo de hidrólise. Esse cultivo produziu as seguintes atividades enzimáticas: 723 U gss-1 de endoglucanases, 2024 U gss-¹ de xilanase, 12,6 U gss-¹ de FPU e 41 U gss-¹ de β-glucosidase. Os resultados da aplicação dessas enzimas na hidrólise indicaram como ótimos 7,0 mL de extrato enzimático (4,2 FPU) e 6 horas para BIN e BOU. As melhores conversões de celulose-glicose foram obtidas para a BOU, atingindo 32,1 % a 65 ºC. Em conclusão, a produção de enzimas no biorreator de leito compactado foi eficiente e o pré-tratamento BOU melhorou a hidrólise da biomassa, aumentando a eficiência de conversão de celulose em glicose.

Fungal cellulases: production by solid-state cultivation in packedbed bioreactor using solid agro-industrial by-products as substrates and application for hydrolysis of sugarcane bagasse / Celulases fúngicas: produção por cultivo sólido em biorreator de leito empacotado usando resíduos sólidos agroindustriais como substratos e aplicação na hidrólise de bagaço de cana

Cellulases are essential for the hydrolysis of lignocellulosic materials in the production of second generation ethanol. Solid-state cultivation is a process that provides high concentrations of enzymes that can be used in this hydrolysis. The objectives of this work were to produce cellulases by cultivating the fungus Myceliophthora thermophila I-1D3b in a packed bed bioreactor with sugarcane bagasse (SCB) and wheat bran (WB) as substrate and to evaluate the efficiency of the enzymatic extract in the hydrolysis of SCB in natura (BIN) and pretreated with ozone, alkali and ultrasound (BOU). The conditions for enzyme production in the bioreactor were SCB:WB at a ratio of 2.3:1 (w/w), 75 % moisture content; 45 ºC; aeration rate 240 L h-¹ and 96 h. The enzyme production was evaluated by endoglucanase, xylanase, filter paper (FPU) and β-glycosidase activities. For the application of the enzymes, a central composed response surface design with 5 repetitions of the central point was used, taking enzyme volume and hydrolysis time as factors. Such cultivation yielded the following enzymatic activities: 723 U gss-¹ of endoglucanases, 2024 U gss-¹ of xylanase, 12.6 U gss-¹ of FPU and 41 U gss-¹ of β-glucosidase. The results of the application tests indicated the best conditions as 7.0 mL of the enzyme extract (4.2 FPU) and 6 hours for BIN and BOU. The best cellulose-glucose conversions were obtained for BOU, reaching 32.1 % at 65 ºC. In conclusion, the enzyme production in the packed bed bioreactor was efficient and BOU pretreatment improved the hydrolysis of biomass, increasing the efficiency of conversion of cellulose to glucose.(AU)

Celulases são essenciais para a hidrólise de materiais lignocelulósicos visando à produção de etanol de segunda geração. O cultivo em estado sólido é um processo que proporciona altas concentrações de enzimas que podem ser aplicadas nessa hidrólise. O objetivo deste trabalho foi produzir celulases pelo fungo Myceliophthora thermophila I-1D3b em biorreator de leito empacotado com bagaço decana (BC) e farelo de trigo (FT) como substrato e avaliar a eficiência deste extrato enzimático na hidrólise de bagaço de cana in natura (BIN) e pré-tratado com ozônio, álcali e ultrassom (BOU). As condições para produção de enzimas no biorreator foram BC:FT numa proporção de 2,3:1 (m/m); 75 %de umidade em base úmida; 45 ºC; taxa de aeração 240 L h-1 e 96 h. A produção de enzimas foi avaliada pelas atividades de endoglucanase, xilanase, papel de filtro (FPU) e β-glicosidase. Para a aplicação das enzimas, foi utilizado um planejamento de superfície de resposta central composto com 5 repetições do ponto central, tendo como fatores o volume de extrato enzimático e o tempo de hidrólise. Esse cultivo produziu as seguintes atividades enzimáticas: 723 U gss-1 de endoglucanases, 2024 U gss-¹ de xilanase, 12,6 U gss-¹ de FPU e 41 U gss-¹ de β-glucosidase. Os resultados da aplicação dessas enzimas na hidrólise indicaram como ótimos 7,0 mL de extrato enzimático (4,2 FPU) e 6 horas para BIN e BOU. As melhores conversões de celulose-glicose foram obtidas para a BOU, atingindo 32,1 % a 65 ºC. Em conclusão, a produção de enzimas no biorreator de leito compactado foi eficiente e o pré-tratamento BOU melhorou a hidrólise da biomassa, aumentando a eficiência de conversão de celulose em glicose.(AU)

Características do trato digestivo, metabolizabilidade e retenção de nutrientes em frangos de corte alimentados com complexo enzimático / Characteristics of the digestive tract, metabolizability and nutrient retention in broilers fed enzymatic complex

The objective was to evaluate the digestive tract characteristics, metabolizability and nutrient retention of broilers fed diets supplemented with enzyme complex (EC). To evaluate the characteristics of the digestive tract 600 female Cobb 500 birds were used, distributed in a completely randomized design, with 5 inclusion levels of the EC (0; 100, 200, 300 and 400 g/ton) and 6 replicates of 20 birds each. To evaluate the metabolizability and the retention of nutrients 200 female Cobb 500 birds at 15 days of age were used, distributed in a completely randomized design with 5 levels of supplementation of the EC and 4 replicates of 10 birds each. No significant effects (P>0.05) were observed for the supplementation of the EC in the intestinal pH, digestive organ weight, intestinal length and metabolizable coefficients of dry matter and crude protein. The metabolizable coefficient of ethereal extract was influenced in a quadratic decreasing form (P<0.01). The metabolizable coefficients of calcium (Ca) and phosphorus (P) were influenced in a quadratic increase (P<0.01), resulting in increased Ca retention in 21.39% and P in 9.56%. Supplementation of the EC in broiler diets improves the metabolizability and retention of P and Ca, without affecting the other parameters evaluated.(AU)

Características do trato digestivo, metabolizabilidade e retenção de nutrientes em frangos de corte alimentados com complexo enzimático / Characteristics of the digestive tract, metabolizability and nutrient retention in broilers fed enzymatic complex

The objective was to evaluate the digestive tract characteristics, metabolizability and nutrient retention of broilers fed diets supplemented with enzyme complex (EC). To evaluate the characteristics of the digestive tract 600 female Cobb 500 birds were used, distributed in a completely randomized design, with 5 inclusion levels of the EC (0; 100, 200, 300 and 400 g/ton) and 6 replicates of 20 birds each. To evaluate the metabolizability and the retention of nutrients 200 female Cobb 500 birds at 15 days of age were used, distributed in a completely randomized design with 5 levels of supplementation of the EC and 4 replicates of 10 birds each. No significant effects (P>0.05) were observed for the supplementation of the EC in the intestinal pH, digestive organ weight, intestinal length and metabolizable coefficients of dry matter and crude protein. The metabolizable coefficient of ethereal extract was influenced in a quadratic decreasing form (P<0.01). The metabolizable coefficients of calcium (Ca) and phosphorus (P) were influenced in a quadratic increase (P<0.01), resulting in increased Ca retention in 21.39% and P in 9.56%. Supplementation of the EC in broiler diets improves the metabolizability and retention of P and Ca, without affecting the other parameters evaluated.(AU)

Production of cellulases by Penicillium oxalicum through solid state fermentation using agroindustrial substrates / Produção de celulases por Penicillium oxalicum em fermentação em estado sólido usando diferentes substratos agroindustriais

The purpose of this study was to define the factors that influence the production of cellulases by Penicillium oxalicum, a strain obtained from a leaf-cutting ant colony and identified based on the ITS gene. The experimental design included solid state fermentation using sugarcane bagasse and lignocellulosic sorghum as the lignocellulosic substrate. The variables were analyzed using a 25-1 fractional factorial design, with three replicates on the central point. All the variables analyzed influenced the production of at least one of the three cellulose types analyzed. The highest values observed were: FPase 4.2 U g-1, CMCase 9.2 U g-1 and Avicelase 8.4 U g-1 using lignocellulosic sorghum as the substrate. The best conditions for enzyme production were: incubation temperature at 40ºC, initial moisture of 60%, pH of 4.0 and a growth period of four days using lignocellulosic sorghum as the substrate.(AU)

O estudo teve como foco a determinação de fatores que influenciam a produção de celulases por uma cepa isolada de ninho de formigas cortadeiras e identificada por meio do gene ITS como Penicillium oxalicum. O processo produtivo foi Fermentação em Estado Sólido utilizando como substrato lignocelulósico bagaço de cana-de-açúcar e sorgo lignocelulósico. As variáveis foram analisadas através de um planejamento fatorial fracionário 25-1, com três repetições no ponto central. Todas as variáveis analisadas influenciaram a produção de pelo menos um dos três tipos de celulases analisados. As maiores atividades observadas foram: FPase 4,2 U g-1; CMCases 9,2 U g-1 e avicelase 8,4 U g-1, utilizando sorgo lignocelulósico como substrato. As melhores condições para produção foram: temperatura de incubação a 40oC, umidade inicial 60%, pH 4,0, tempo de cultivo de quatro dias, utilizando como substrato sorgo lignocelulósico.(AU)

Enzymatic saccharification and fermentation of cellulosic date palm wastes to glucose and lactic acid

The bioconversion of cellulosic wastes into high-value bio-products by saccharification and fermentation processes is an important step that can reduce the environmental pollution caused by agricultural wastes. In this study, enzymatic saccharification of treated and untreated date palm cellulosic wastes by the cellulases from Geobacillus stearothermophilus was optimized. The alkaline pre-treatment of the date palm wastes was found to be effective in increasing the saccharification percentage. The maximum rate of saccharification was found at a substrate concentration of 4% and enzyme concentration of 30 FPU/g of substrate. The optimum pH and temperature for the bioconversions were 5.0 and 50 °C, respectively, after 24 h of incubation, with a yield of 31.56 mg/mL of glucose at a saccharification degree of 71.03%. The saccharification was increased to 94.88% by removal of the hydrolysate after 24 h by using a two-step hydrolysis. Significant lactic acid production (27.8 mg/mL) was obtained by separate saccharification and fermentation after 72 h of incubation. The results indicate that production of fermentable sugar and lactic acid is feasible and may reduce environmental pollution by using date palm wastes as a cheap substrate. (AU)

Cellulase activity and dissolved organic carbon release from lignocellulose macrophyte-derived in four trophic conditions

Considering the importance of lignocellulose macrophyte-derived for the energy flux in aquatic ecosystems and the nutrient concentrations as a function of force which influences the decomposition process, this study aims to relate the enzymatic activity and lignocellulose hydrolysis in different trophic statuses. Water samples and two macrophyte species were collected from the littoral zone of a subtropical Brazilian Reservoir. A lignocellulosic matrix was obtained using aqueous extraction of dried plant material (40 °C). Incubations for decomposition of the lignocellulosic matrix were prepared using lignocelluloses, inoculums and filtered water simulating different trophic statuses with the same N:P ratio. The particulate organic carbon and dissolved organic carbon (POC and DOC, respectively) were quantified, the cellulase enzymatic activity was measured by releasing reducing sugars and immobilized carbon was analyzed by filtration. During the cellulose degradation indicated by the cellulase activity, the dissolved organic carbon daily rate and enzyme activity increased. It was related to a fast hydrolysable fraction of cellulose that contributed to short-term carbon immobilization (ca. 10 days). After approximately 20 days, the dissolved organic carbon and enzyme activity were inversely correlated suggesting that the respiration of microorganisms was responsible for carbon mineralization. Cellulose was an important resource in low nutrient conditions (oligotrophic). However, the detritus quality played a major role in the lignocelluloses degradation (i.e., enzyme activity) and carbon release.(AU)

Enhanced alkaline cellulases production by the thermohalophilic Aspergillus terreus AUMC 10138 mutated by physical and chemical mutagens using corn stover as substrate

Abstract A thermohalophilic fungus, Aspergillus terreus AUMC 10138, isolated from the Wadi El-Natrun soda lakes in northern Egypt was exposed successively to gamma and UV-radiation (physical mutagens) and ethyl methan-sulfonate (EMS; chemical mutagen) to enhance alkaline cellulase production under solid state fermentation (SSF) conditions. The effects of different carbon sources, initial moisture, incubation temperature, initial pH, incubation period, inoculum levels and different concentrations of NaCl on production of alkaline filter paper activity (FPase), carboxymethyl cellulase (CMCase) and β-glucosidase by the wild-type and mutant strains of A. terreus were evaluated under SSF. The optimum conditions for maximum production of FPase, CMCase and β-glucosidase were found to be the corn stover: moisture ratio of 1:3(w/v), temperature 45 °C, pH range, 9.0–11.0, and fermentation for 4, 4 and 7 day, respectively. Inoculum levels of 30% for β-glucosidase and 40% for FPase, CMCase gave the higher cellulase production by the wild-type and mutant strains, respectively. Higher production of all three enzymes was obtained at a 5% NaCl. Under the optimized conditions, the mutant strain A. terreus M-17 produced FPase (729 U/g), CMCase (1,783 U/g), and β-glucosidase (342 U/g), which is, 1.85, 1.97 and 2.31-fold higher than the wild-type strain. Our results confirmed that mutant strain M-17 could be a promising alkaline cellulase enzyme producer employing lignocellulosics especially corn stover.(AU)

Caracterização de fungos isolados da região Amazônica quanto ao potencial para produção das enzimas envolvidas na conversão da biomassa vegetal / Characterization of fungi isolated from the Amazon region for the potential of biomass-degrading enzymes production

A conversão da biomassa vegetal proveniente de resíduos agroindustriais e florestais em biocombustíveis e bioprodutos, dentro do conceito de biorrefinarias, é de grande interesse, principalmente para o Brasil, onde a agroenergia possui um enorme potencial de desenvolvimento. Entretanto, para garantir a viabilidade do processo de conversão, é fundamental reduzir o custo das enzimas utilizadas na etapa de hidrólise. Para isso, deve-se dispor da peça chave deste processo, que é o microrganismo. Nesse contexto, o objetivo deste trabalho foi avaliar fungos isolados da região Amazônica em relação ao potencial de produção das enzimas celulases e xilanases. De um total de 40 isolados cultivados por fermentação em estado sólido (FES), durante 10 dias, os fungos que se destacaram quanto à produção de endoglucanase (351,79Ug-1 em 120h) e -glicosidase (62,31Ug-1em 72h) foi o P47C3 (A. niger), e na produção de xilanase (1076,94Ug-1 em 72h) e FPase (2,46Ug-1 em 120h) foram o P6B2 (A. oryzae) e o P40B3, respectivamente. Os resultados obtidos demonstram o enorme potencial de aplicação das enzimas produzidas pelos fungos isolados da Amazônia, contribuindo, assim, para gerar os avanços tecnológicos necessários para o aumento da eficiência do uso da biomassa vegetal como fonte de energia renovável.(AU)

The conversion of biomass from forestry and agroindustrial residues into biofuels and bioproducts, within the biorefinery concept, is of great interest, especially to Brazil, where bioenergy has a huge potential for development. However, to ensure the viability of the conversion process it is essential to reduce the cost of the enzymes used in the hydrolysis step. For this, one must have the key element of this process, which is the microorganism. In this context, the objective of this study was to evaluate different fungi isolated from the Amazon region for their potential in terms of the production of cellulase and xylanase enzymes. Of a total of 40 strains cultivated under solid state fermentation (SSF) for 10 days, the strain that stood out for the production of endoglucanase (351.79Ug-1120h) and -glucosidase (62.31Ug-1 at 72h) was P47C3 (A. niger) whereas for xylanase (1076.94Ug-1 in 72 hours) and FPase (2.46Ug-1 in120 hours) were P6B2 (A. oryzae) and P40B3, respectively. These results demonstrate the great potential application of the enzymes produced by the Amazon isolated fungi, thus contributing to generate the necessary technological advances in order to increase the efficiency of the use of biomass as a renewable energy source.(AU)

Estratégias para produção de celulases através de fermentação submersa utilizando fungos isolados do bioma brasileiro / Strategies to increase cellulase production with submerged fermentation using fungi isolated from the Brazilian biome

fermentação submersa utilizando fungos isolados do bioma brasileiro RESUMO. Estudos sobre novas fontes microbianas e análises mais acuradas das etapas que compõem a produção de celulases são essenciais como estratégias para diminuir os custos gerados pelo uso de celulases nos processos de obtenção de açúcares fermentescíveis. O trabalho teve como objetivo a seleção de fungos filamentosos produtores de celulases e a investigação de parâmetros que envolvem a produção enzimática em fermentação submersa. O primeiro teste consistiu em selecionar os melhores fungos produtores de celulases totais em frascos Erlenmeyer contendo 200 mL de meio de cultura específico. O teste subsequente teve o intuito de investigar a produção enzimática com quatro tipos de açúcares solúveis: glicose, lactose, sacarose e xilose. Nos testes em biorreator foram analisados três diferentes estratégias de inoculação. Na etapa de seleção a melhor atividade de FPase foi apresentada por Trichoderma sp. CMIAT 041 (49,9 FPU L-1) e CMCase pelo fungo Lasiodiplodia theobromae CMIAT 096 (350,0 U L-1). O uso de sacarose mostrou-se ser a melhor opção dentre os açúcares solúveis testados, apresentando os maiores valores de atividade de FPase (49,9 FPU L-1) e CMCase (119,7 U L-1). A melhor estratégia de inoculação foi a suspensão de esporos obtidos a partir de fermentação em farelo de trigo, no tempo 72h.(AU)

Studies on new microbial sources of cellulase and accurate assessment of the steps that increase cellulase production are essential strategies to reduce costs of various processes using such enzymes. This study aimed at the selection of cellulase-producing filamentous fungi, and at the research of parameters involving cellulase production by submerged fermentation. The first test consisted of selecting the best cellulase-producing microorganisms (FPase) in Erlenmeyer flasks containing 200 mL of specific growth medium. The next test was designed to further investigate the enzyme production in fermentation with four types of soluble sugars: glucose, lactose, sucrose and xylose. In bioreactor tests, three different inoculation strategies were analyzed. The best FPase activity was presented by the strain Trichoderma sp. CMIAT 041 (49.9 FPU L-1) and CMCase by the fungus Lasiodiplodia theobromae CMIAT 096 (350.0 U L-1). Sucrose proved to be the best option among the soluble sugars tested, with higher rates of FPase activity (49.9 FPU L-1) and CMCase (119.7 U L-1). The best inoculation strategy for the bioreactor was a spore suspension obtained from a semi-solid state fermentation of wheat bran for 72h.(AU)

Influence of nitrogen sources on the enzymatic activity and grown by Lentinula edodes in biomass Eucalyptus benthamii / Influência das fontes de nitrogênio na atividade enzimática e crescimento de Lentinula edodes na biomassa Eucalyptus benthamii

Lignocellulose is the most abundant environmental component and a renewable organic resource in soil. There are some filamentous fungi which developed the ability to break down and use cellulose, hemicellulose and lignin as an energy source. The objective of this research was to analyze the effect of three nitrogen resources (ammonium sulfate, saltpetre, soybean) in the holocellulolitic activity of Lentinula edodes EF 50 using as substrate sawdust E. benthamii. An experimental design mixture was applied with repetition in the central point consisting of seven treatments (T) of equal concentrations of nitrogen in ammonium sulfate, potassium nitrate and soybean. The enzymatic activity of avicelase, carboxymetilcellulase, β-glucosidase, xylanases and manganese peroxidase was determined. The humidity, pH, water activity (aw) and qualitative analysis of mycelial growth in 8 times of cultivation were evaluated. The results showed negative effect on enzyme production in treatments with maximum concentration of ammonium sulfate and potassium nitrate. The treatments with cooked soybean flour expressed higher enzymatic activities in times of 3, 6 and 9 days of culture, except in the activity of manganese peroxidase. The highest production was observed in the treatment with ammonium sulfate, and soybean (83.86 UI.L–1) at 20 days of cultivation.(AU)

Lignocelulose é o componente mais abundante do meio ambiente e recurso orgânico renovável no solo. Alguns fungos filamentosos têm desenvolvido a habilidade de degradar e utilizar celulose, hemicelulose e lignina como fonte de energia. O objetivo deste trabalho foi analisar o efeito de três fontes de nitrogênio (sulfato de amônio, nitrato de potássio e farelo de soja) na atividade enzimática de Lentinula edodes EF 50 utilizando como substrato serragem de E. benthamii. Foi aplicado um planejamento experimental de mistura com três repetições no ponto central constituído de sete tratamentos (T) de iguais concentrações em nitrogênio de sulfato de amônia, nitrato de potássio e farinha de soja cozida. Foram determinadas a atividade enzimática da avicelase, carboximetilcelulase, β-glicosidase, xilanases e manganês peroxidase. Foram avaliados o teor de umidade, pH, atividade de água (aw) e análise qualitativa do crescimento micelial em 8 tempos de cultivo. Os resultados mostraram efeito negativo na produção das enzimas nos tratamentos com máxima concentração de sulfato de amônia e nitrato de potássio. Os tratamentos com farinha de soja cozida expressaram maiores atividades enzimáticas, nos tempos de 3, 6 e 9 dias de cultivo exceto na atividade do manganês peroxidase. A maior produção foi observada no tratamento com sulfato de amônia e farinha de soja cozida (83.86 UI.L–1) em 20 dias de cultivo.(AU)

Extracellular production of avicelase by the thermophilic soil bacterium Bacillus sp. SMIA-2 / Produção de avicelase extracelular pela bactéria termofílica Bacillus sp. SMIA-2

Nowadays, the isolation of new bacterial strains that produce enzymes with novel properties is a subject of great relevance to the scientific community. This study, in order to search for producers of new cellulase strains, investigated the avicelase production by thermophilic Bacillus sp. strain SMIA-2. The best avicelase activity was observed in a culture medium containing 0.5% (w v-1) avicel and 0.5% (w v-1) corn steep liquor with initial pH 7.5-8.0 incubated at 50oC. When avicel was replaced in the medium by the treated sugarcane bagasse (0.5%, w v-1) the avicelase activity levels were not affected. Studies on the avicelase characterization revealed that the optimum pH of the enzyme was found to be 8.5 and the enzyme retained more than 80% of its activity after incubation at room temperature for 2h at pH 6.5-8.5. The optimum temperature of this enzyme was 70oC and the enzyme retained 67% of the original activity after 20 min. of heat treatment at 70oC. Avicelase was stimulated by Mn2+ and Co2+, whereas Hg2+ greatly inhibited the enzyme activity.(AU)

Atualmente, o isolamento de estirpes de bactérias que produzem enzimas com novas propriedades é um tema de grande relevância para a comunidade científica. Este trabalho, buscando por novas cepas produtoras de celulases, investigou a produção de avicelases pelo termofílico Bacillus sp. cepa SMIA-2. A melhor atividade da enzima foi obtida em uma cultura contendo 0,5% (p v-1) avicel e 0,5% (p v-1) água de maceração de milho com pH inicial de pH 7,5-8,0 e incubada a 50oC. A substituição da avicel no meio de cultura pelo bagaço de cana- de- açúcar tratado (0,5%, p v-1) não afetou os níveis de atividade da avicelase. Estudos sobre a caracterização da avicelase revelaram que o pH para atividade ótima da enzima foi 8,5 e que a mesma reteve mais de 80% de sua atividade após ser incubada à temperatura ambiente por 2 h a pH 6,5- 8,5. A temperatura ótima da avicelase foi 70oC e a enzima reteve 67% da sua atividade original após 20 min. de incubação a 70oC. A avicelase foi estimulada pelos íons Mn2+ e Co2+, ao passo que Hg2+ inibiu a atividade da enzima.(AU)