Consumo e digestibilidade aparente total em bovinos sob suplementação com enzimas fibrolíticas

Avaliou-se o efeito da suplementação com enzimas fibrolíticas (celulase e xilanase) sobre o consumo e a digestibilidade aparente total dos nutrientes de dietas compostas de silagem de milho e feno de tifton 85 (Cynodon spp.) cortado aos 90 dias. Oito bovinos (340 kg de PV) fistulados no rúmen foram distribuídos em dois quadrados latinos 4 x 4, em esquema fatorial 2 x 2 (fonte de volumoso e adição ou não de enzimas). O complexo enzimático testado foi proveniente de fonte comercial e extraído dos fungos Aspergillus niger e Trichoderma longibrachiatum, sendo fornecido na quantidade de 12 g/animal/dia, misturado à ração total. Não houve efeito da suplementação enzimática sobre o consumo de nutrientes para ambos os volumosos, porém, a adição de enzimas aumentou a digestibilidade total de FDN, FDA e CEL de 36,87; 36,21 e 46,89%, respectivamente, para 41,19; 40,01 e 50,46%, respectivamente.

Estudo da influência de pré-tratamentos de dois resíduos lignocelulósicos (bagaço do pedúnculo de caju e casca de coco) utilizados como substrato na indução à síntese de enzimas celulolíticas

Nowadays generation ethanol second, that t is obtained from fermentation of sugars of hydrolyses of cellulose, is gaining attention worldwide as a viable alternative to petroleum mainly for being a renewable resource. The increase of first generation ethanol production i.e. that obtained from sugar-cane molasses could lead to a reduction of lands sustainable for crops and food production. However, second generation ethanol needs technologic pathway for reduce the bottlenecks as production of enzymes to hydrolysis the cellulose to glucose i.e. the cellulases as well as the development of efficient biomass pretreatment and of low-cost. In this work Trichoderma reesei ATCC 2768 was cultivated under submerged fermentation to produce cellulases using as substrates waste of lignocellulosic material such as cashew apple bagasse as well as coconut bagasse with and without pretreatment. For pretreatment the bagasses were treated with 1 M NaOH and by explosion at high pressure. Enzyme production was carried out in shaker (temperature of 27ºC, 150 rpm and initial medium pH of 4.8). Results showed that T.reesei ATCC 2768 showed the higher cellulase production when the cashew apple bagasse was treated with 1M NaOH (2.160 UI/mL of CMCase and 0.215 UI/mL of FPase), in which the conversion of cellulose, in terms of total reducing sugars, was of 98.38%, when compared to pretreatment by explosion at high pressure (0.853 UI/mL of CMCase and 0.172 UI/mL of Fpase) showing a conversion of 47.39% of total reducing sugars. Cellulase production is lower for the medium containing coconut bagasse treated with 1M NaOH (0.480 UI/mL of CMcase and 0.073 UI/mL of FPase), giving a conversion of 49.5% in terms of total reducing sugars. Cashew apple bagasse without pretreatment showed cellulase activities lower (0.535 UI/mL of CMCase and 0,152 UI/mL of FPase) then pretreated bagasse while the coconut bagasse without pretreatment did not show any enzymatic activity. Maximum cell concentration was obtained using cashew nut bagasse as well as coconut shell bagasse treated with 1M NaOH, with 2.92 g/L and 1.97 g/L, respectively. These were higher than for the experiments in which the substrates were treated by explosion at high pressure, 1.93 g/L and 1.17 g/L. Cashew apple is a potential inducer for cellulolytic enzymes synthysis showing better results than coconut bagasse. Pretreatment improves the process for the cellulolytic enzyme production

Produção de enzimas extracelulares por Fusarium solani de maracujazeiro amarelo

Uma das principais doenças do maracujazeiro, na maioria dos estados produtores do Brasil, é a podridão do colo, causada por Fusarium solani. Pouco se sabe a respeito da fisiologia deste patógeno do maracujazeiro amarelo, principalmente quanto à produção de enzimas extracelulares. O objetivo do presente trabalho foi verificar, em meios de cultura individuais e apropriados, a produção das enzimas extracelulares amilase, lipase, celulase, proteases (caseinase e gelatinase), lacase (oxidase) e catalase por isolados de F. solani, provenientes de maracujazeiro amarelo. O delineamento experimental adotado foi o inteiramente casualizado, em esquema de dois fatores (nove isolados versus sete enzimas), com três repetições. Todos os isolados de F. solani produziram, de maneira semiquantitativa, as enzimas extracelulares amilase, lipase, celulase, caseinase (protease) e lacase (oxidase). No entanto, a quantidade produzida de cada enzima foi significativamente diferente entre os isolados. As enzimas extracelulares gelatinase (protease) e catalase foram produzidas em pouca quantidade e de maneira igual por todos os isolados do fungo.

Produção de enzimas por fungos em fermentação semi-sólida utilizando bagaço de coco e pedúnculo de caju como substratos

The production of enzymes by microorganisms using organic residues is important and it can be associated with several applications such as food and chemical industries and so on. The objective of this work is the production of CMCase, Xylanase, Avicelase and FPase enzymes by solid state fermentation (SSF) using as substrates: bagasse of coconut and dried cashew stem. The microorganisms employed are Penicillium chrysogenum and an isolated fungus from the coconut bark (Aspergillus fumigatus). Through the factorial design methodology and response surface analysis it was possible to study the influence of the humidity and pH. For Penicillium chrysogenum and the isolated fungus, the coconut bagasse was used as culture medium. In another fermentation, it was used the mixture of coconut bagasse and cashew stem. Fermentations were conducted using only the coconut bagasse as substrate in cultures with Penicillium chrysogenum fungus and the isolated one. A mixture with 50% of coconut and 50% of cashew stem was employed only for Penicillium chrysogenum fungus, the cultivation conditions were: 120 hours at 30 °C in BOD, changing humidity and pH values. In order to check the influence of the variables: humidity and pH, a 2 2 factorial experimental design was developed, and then two factors with two levels for each factor and three repetitions at the central point. The levels of the independent variables used in ascending order (-1, 0, +1), to humidity, 66%, 70.5% and 75% and pH 3, 5 and 7, respectively. The software STATISTICA TM (version 7.0, StatSoft, Inc.) was used to calculate the main effects of the variables and their interactions. The response surface methodology was used to optimize the conditions of the SSF. A chemical and a physic-chemical characterization of the coconut bagasse have determined the composition of cellulose (%) = 39.09; Hemicellulose (%) = 23.80, Total Lignin (%) = 36.22 and Pectin (%) = 1.64. To the characterization of cashew stem, the values were cellulose (g) = 15.91 Hemicellulose (%) = 16.77, Total Lignin (%) = 30.04 and Pectin (%) = 15.24. The results indicate the potential of the materials as substrate for semisolid fermentation enzyme production. The two microorganisms used are presented as good producers of cellulases. The results showed the potential of the fungus in the production of CMCase enzyme, with a maximum of 0.282 UI/mL and the Avicelase enzyme the maximum value ranged from 0.018 to 0.020 UI/ mL, using only coconut bagasse as substrate. The Penicillium chrysogenum fungus has showed the best results for CMCase = 0.294 UI/mL, FPase = 0.058 UI/mL, Avicelase = 0.010 UI/mL and Xylanase = 0.644 UI/ mL enzyme, using coconut bagasse and cashew stem as substrates. The Penicllium chrysogenum fungus showed enzymatic activities using only the coconut as substrate for CMCase = 0.233 UI/mL, FPase = 0.031 to 0.032 UI/ mL, Avicelase = 0.018 to 0.020 UI/mL and Xylanase = 0.735 UI/ mL. Thus, it can be concluded that the used organisms and substrates have offered potential for enzyme production processes in a semi-solid cultivation

Palma forrageira (Opuntia ficus indica e Nopalea cochenillifera) como matéria-prima para produção de etanol celulósico e enzimas celulolíticas

The need for new sources of energy and the concern about the environment have pushed the search for renewable energy sources such as ethanol. The use of lignocellulosic biomass as substrate appears as an important alternative because of the abundance of this raw material and for it does not compete with food production. However, the process still meets difficulties of implementation, including the cost for production of enzymes that degrade cellulose to fermentable sugars. The aim of this study was to evaluate the behavior of the species of cactus pear Opuntia ficus indica and Nopalea cochenillifera, commonly found in northeastern Brazil, as raw materials for the production of: 1) cellulosic ethanol by simultaneous saccharification and fermentation (SSF) process, using two different strains of Saccharomyces cerevisiae (PE-2 and LNF CA-11), and 2) cellulolytic enzymes by semi-solid state fermentation (SSSF) using the filamentous fungus Penicillium chrysogenum. Before alcoholic fermentation process, the material was conditioned and pretreated by three different strategies: alkaline hydrogen peroxide, alkaline using NaOH and acid using H2SO4 followed by alkaline delignification with NaOH. Analysis of composition, crystallinity and enzymatic digestibility were carried out with the material before and after pretreatment. In addition, scanning electron microscopy images were used to compare qualitatively the material and observe the effects of pretreatments. An experimental design 2² with triplicate at the central point was used to evaluate the influence of temperature (30, 40 and 45 °C) and the initial charge of substrate (3, 4 and 5% cellulose) in the SSF process using the material obtained through the best condition and testing both strains of S. cerevisiae, one of them flocculent (LNF CA-11). For cellulase production, the filamentous fungus P. chrysogenum was tested with N. cochenillifera in the raw condition (without pretreatment) and pretrated hydrothermically, varying the pH of the fermentative medium (3, 5 and 7). The characterization of cactus pear resulted in 31.55% cellulose, 17.12% hemicellulose and 10.25% lignin for N. cochenillifera and 34.86% cellulose, 19.97% hemicellulose and 15.72% lignin for O. ficus indica. It has also been determined, to N. cochenillifera and O. ficus indica, the content of pectin (5.44% and 5.55% of calcium pectate, respectively), extractives (26.90% and 9.69%, respectively) and ashes (5.40% and 5.95%). Pretreatment using alkaline hydrogen peroxide resulted in the best cellulose recovery results (86.16% for N. cochenillifera and 93.59% for O. ficus indica) and delignification (48.79% and 23.84% for N. cochenillifera and O. ficus indica, respectively). This pretreatment was also the only one which did not increase the crystallinity index of the samples, in the case of O. ficus indica. However, when analyzing the enzymatic digestibility of cellulose, alkali pretreatment was the one which showed the best yields and therefore it was chosen for the tests in SSF. The experiments showed higher yield of conversion of cellulose to ethanol by PE-2 strain using the pretreated N. cochenillifera (93.81%) at 40 °C using 4% initial charge of cellulose. N. cochenillifera gave better yields than O. ficus indica and PE-2 strain showed better performance than CA-11. N. cochenillifera proved to be a substrate that can be used in the SSSF for enzymes production, reaching values of 1.00 U/g of CMCase and 0.85 FPU/g. The pretreatment was not effective to increase the enzymatic activity values

Associação entre polimorfismos em enzimas de reparo de DNA e ocorrência de meningite bacteriana.

Despite advances in vaccine development and therapy, bacterial meningitis (BM) remains a major cause of death and long-term neurological disabilities. As part of the host inflammatory response to the invading pathogen, factors such as reactive oxygen species are generated, which may damage DNA and trigger the overactivation of DNA repair mechanisms. It is conceivable that the individual susceptibility and outcome of BM may be in part determined by non synonymous polymorphisms that may alter the function of crucial BER DNA repair enzymes as PARP-1, OGG-1 and APE-1. These enzymes, in addition to their important DNA repair function, also perform role of inflammatory regulators. In this work was investigated the non synonymous SNPs APE-1 Asn148Glu, OGG-1 Ser326Cys,PARP-1 Val762Ala, PARP-1 Pro882Leu and PARP-1 Cys908Tyr in patients with bacterial meningitis (BM), chronic meningitis (CM), aseptic meningitis (AM) and not infected (controls). As results we found increased frequency of variant alleles of PARP-1 Val762Ala (P = 0.005) and APE-1 Asn148Glu (P=0.018) in BM patients, APE-1 Asn148Glu in AM patients (P = 0.012) and decrease in the frequency of the variant allele OGG-1 Ser326Cys in patients with CM (P = 0.013), regarding the allelic frequencies in the controls. A major incidence of individuals heterozygous and/ or polymorphic homozygous in BM for PARP-1 Val762Ala (P= 0.0399, OD 4.2, 95% IC 1.213 -14.545) and PARP-1 Val762Ala/ APE-1 Asn148Glu (P = 0.0238, OD 11.111, 95% IC 1.274 - 96.914) was observed related to what was expected in a not infected population. It was also observed a major incidence of combined SNPs in the BM patients compared with the control group (P=0.0281), giving evidences that SNPs can cause some susceptibility to the disease. This combined effect of SNPs seems to regulate the principal cytokines and other factors related to BM inflammatory response and point the importance of DNA repair not only to repair activity when DNA is damaged, but to others essential functions to human organism balance.

Relação entre padrão comportamental, estágios do ciclo de muda e atividade de enzimas digestivas proteolíticas em juvenis do camarão marinho Litopenaeus vannamei (Crustacea: Penaeidae)

Shrimp farming in Brazil is a consolidated activity, having brought economical and social gains to several states with the largest production concentrated in the northeast. This fact is also reflected in higher feed intake, necessitating a more efficient feed management. Currently, management techniques already foresee food loss due to molting. In this sense, studies relating shrimp s digestive physiology, molting physiology and behavioral response of shrimp feed can optimize the feed management. Thus, our study aimed to evaluate the behavioral response of the marine shrimp L. vannamei (Crustacea: Penaeidae) in accordance with the stages of moulting cycle and feeding schedules based on higher or lower activity of proteolytic digestive enzymes; also, to investigate the influence of feeding schedule on hepatosomatic index and non-specific and specific protease activity (trypsin). Experiments were carried out at the Laboratory of Shrimp Behavioral Studies at UFRN in partnership with the Laboratory of Enzimology UFPE. Juveniles of L. vannamei weighting 5.25 g (+ 0.25 g) were kept in aquaria at a density of 33 shrimp m -2. In the first experiment, shrimp were fed in the light phase or in the dark phase for 8 days; in the ninth day, the animals were observed for 15 minutes every hour during the 12 hours of each phase of the photoperiod. We recorded the frequency of inactivity, exploration, food intake, burrowing, swimming and crawling behavior. At the end of the 12th observation session, the shrimp were sacrified and classified by the method of setogenesis in the molt cycle stages A, B, C, D0, D1, D2 or D3. We found that the shrimp in A stage show high levels of inactivity. Moreover, the frequency of food intake was very low. The shrimp in D3 stage also had low food intake and high inactivity associated with elevated frequencies of burrowing. In the second experiment, shrimp were kept in physiological acclimation to experimental conditions for 28 days, distributed in 12 treatments in the light phase and 12 treatments in the dark phase. In the end, the animals were sacrified and dissected to assess non-specific and specific protease activity (trypsin) activity. In general, these parameters did not vary among animals fed in the light phase and those fed in the dark phase. However, significant differences were found in the activity of specific and nonspecific proteases in relation to food treatment. In the light phase, the major proteolytic activities converged to 10 hours after the start of the light phase, while the lowest activities converged to 6 hours after the beginning of this phase. In the dark phase, the highest enzyme activity converged to 12 hours after the onset of phase, while the lowest activities converged to 3 hours after the onset of phase. In the third experiment, we sought to evaluate the behavioral responses of shrimp in relation to dietary treatments based on higher or lower activity of proteolytic enzymes, considering the results of the second experiment. The behavioral categories observed were the same as the ones in the first experiment, with observations of 30 minutes (15min before and 15min after food supply). We found variation in behavioral responses as a function of the treatments, with greater intake of food in shrimp fed during the period of greatest activity of proteolytic enzymes, in the light phase. Thus we see that periodic events associated with the shrimp s physiology interfere in their behavioral responses, revealing situations that are more adjustable to the provision of food, and consequently optimizing feeding management

Níveis séricos de enzimas de função hepática em frangos de corte de criação industrial clinicamente saudáveis

The values for the main hepatic enzymes included in the profiles of screen clinical biochemistry, alanine-aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (FA), lactate desidrogenase (LDH) and gamaglutamiltransferase (GGT), in samples of serum of broiler chickens in industrial system, clinically healthy, starting from the seventh day of life, until the slaughter (42 days) in weekly intervals were determined. Significant variations were not observed in the analyses in relation to the age of the birds for none of the appraised enzymes.

Concentração de enzimas amilolíticas na hidrólise do amido de gengibre

Uma possibilidade de incremento na cadeia produtiva do gengibre seria o uso de rizomas desclassificados para comercialização in natura, ou mesmo o resíduo da extração de óleos, como matérias-primas para a obtenção de bebidas destiladas. O amido não é diretamente fermentável, necessitando de uma hidrólise prévia de suas cadeias para a obtenção de glicose. Neste trabalho, objetivou-se avaliar o efeito das concentrações de amilases sobre o perfil de açúcares e rendimento no processo de hidrólise-sacarificação de gengibre. O processo seguiu o delineamento central composto rotacional para dois fatores, totalizando 11 tratamentos. Os resultados obtidos mostram o efeito das concentrações da α-amilase (Termamyl 2X) e da amiloglucosidase (AMG 300L) sobre o teor de glicose do hidrolisado e efeito da concentração de amiloglucosidase sobre o teor de dextrina. Os maiores rendimentos de hidrólise da suspensão de gengibre foram obtidos nas condições de elevada concentração de -amilase e amiloglucosidase.

Atividade das enzimas invertases e acúmulo de sacarose em cana-de-açúcar sob efeito do nitrato de potássio, etefon e etil-trinexapac

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

Atividade de enzimas oxidativas do Lentinula edodes em substratos agroindustriais

Basidyomycete Lentinula edodes has its related enzymatic activity mainly on the agribusiness waste kind used as a substrate. The objective of this work was to verify the activity of oxidative enzymes lacase (Lac), lignina peroxidase (LiP) and manganese peroxidase (MnP) of three L. edodes strains, in stationary system, cultivated the 25 degrees C, in the absence of light, in substrates wtih 20% of bran of rice, 1% of CaCO(3) and 79% of rice husk (CA), eucalyptus sawdust (SE), cassava bagasse (BM) and sugarcane bagasse (BC), adjusted to 60% of humidity. The Lac and MnP activities were bigger in eucalyptus sawdust (SE) and sugarcane bagasse (BC). The UP activity was not induced for tested substrates. The rice husk (CA) and cassava bagasse (BM) Substrates, although are not adequate to produce Lac or MnP, can be used as additives to increase the porosity, air availability and easy metabolism polysaccharides.

Enzimas termoestáveis: fontes, produção e aplicação industrial

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

Meta-análise da digestibilidade ileal de aminoácidos e minerais em suínos alimentados com dietas contendo enzimas

O objetivo deste trabalho foi avaliar, por meio da meta-análise, o efeito da fitase e da xilanase sobre a digestibilidade ileal aparente (DIa) de aminoácidos, cálcio e fósforo, em suínos em fase de crescimento. A base de dados consistiu de 21 artigos publicados entre 1998 e 2009, no total de 82 tratamentos e 644 suínos. A meta-análise foi realizada por análise gráfica, de correlação, de variância-covariância. As concentrações de fósforo fítico e as frações fibra em detergente neutro, fibra em detergente ácido e lignina em detergente ácido, nas dietas, apresentaram correlações baixas e negativas com a DIa do cálcio, fósforo e aminoácidos. A adição de fitase às dietas aumentou em 2% a DIa da arginina, em 14% a do cálcio e em 34% a do fósforo. A DIa da arginina, fenilalanina, isoleucina e lisina foi 3,3% superior em suínos alimentados com dietas com xilanase, em relação às dietas sem a enzima. O fósforo fítico e as fibras, nas dietas, reduzem a DIa do cálcio, do fósforo e dos aminoácidos essenciais. O uso de fitase e xilanase, nas dietas, melhora o aproveitamento de cálcio, fósforo e alguns aminoácidos. No entanto, o excesso de cálcio e fósforo nas dietas reduz a ação da fitase sobre a digestibilidade ileal dos nutrientes.

Avaliação da celulase e pectinase como enzimas complementares, no processo de hidrólise-sacarificação do farelo de mandioca para produção de etanol

Neste trabalho objetivou-se avaliar o uso de enzimas complementares no processo enzimático de hidrólise e sacarificação para a produção de etanol a partir do resíduo fibroso das fecularias. Os resultados obtidos demonstraram que 63,42% do amido foram hidrolisados no tratamento em que não se utilizaram enzimas complementares. No tratamento com as duas enzimas complementares foram hidrolisados 89,55%, no tratamento com celulase 65,42% e no tratamento com pectinase 88,73%. A prensagem do resíduo após o processo de hidrólise e sacarificação mostrou-se eficiente, ficando 10,43% do total de açúcares obtidos retidos no resíduo fibroso final. Portanto, o tratamento em que se utilizou a pectinase como enzima complementar na hidrólise foi o melhor. A celulase não apresentou efeito significativo no rendimento do processo.

Suplementação de enzimas exógenas em dieta microparticulada para larvicultura do pacu

Este experimento foi realizado para avaliar os efeitos da suplementação de enzimas exógenas (pancreatina suína) em microdietas sobre o crescimento, a sobrevivência e as alterações morfológicas do trato digestório de larvas de pacu, Piaractus mesopotamicus. Foram testados oito programas alimentares: alimentação exclusiva com náuplios de Artemia (AV); alimentação exclusiva com dieta microparticulada com (DMP) ou sem (DM) suplementação enzimática; substituição, aos cinco dias, dos náuplios de Artemia por dietas inertes com (AV5DMP) ou sem (AV5DM) suplementação; e substituição dos náuplios aos dez dias por dietas com (AV10DMP) ou sem (AV10DM) suplementação. O experimento teve duração de 28 dias. Larvas que receberam o alimento vivo durante todo o período experimental apresentaram maiores médias de peso. O efeito negativo da supressão do alimento vivo sobre o crescimento das larvas foi verificado tanto na substituição aos cinco dias como aos dez dias. No entanto, nas avaliações biométricas subseqüentes, observaram-se efeitos positivos da suplementação enzimática; a partir do 20º dia de experimento, as larvas que receberam a dieta suplementada com enzima exógena apresentaram peso médio estatisticamente superior ao daquelas alimentadas com a dieta sem suplementação. As diferenças morfológicas mais evidentes proporcionadas pela suplementação enzimática foram observadas nas larvas que receberam substituição alimentar aos cinco dias. As diferenças foram relativas à quantidade de grânulos de zimogênio no pâncreas e às inclusões supranucleares no intestino. As larvas submetidas à transição alimentar aos dez dias de experimento já apresentavam diferenciação morfológica do sistema digestório mais avançada, assemelhando-se muito às larvas do tratamento com alimento vivo. Os resultados deste experimento indicam que a suplementação com pancreatina proporcionou efeitos positivos sobre o crescimento e a sobrevivência das larvas de pacu.