Agricultural by-products such as wheat straw are attractive feedstocks for the production of second generation bioethanol due to their high abundance. However, the presence of lignin in these lignocellulosic materials hinders the enzymatic hydrolysis of cellulose. The purpose of this work is to study the ability of a laccase-mediator system to remove lignin improving saccharification, as a pretreatment of wheat straw, and to analyze the chemical modifications produced in the remaining lignin moiety. Up to 48% lignin removal from ground wheat straw was attained by pretreatment with Pycnoporus cinnabarinus laccase and 1-hydroxybenzotriazole (HBT) as mediator, followed by alkaline peroxide extraction. The lignin removal directly correlated with increases (~60%) in glucose yields after enzymatic saccharification. The pretreatment using laccase alone (without mediator) removed up to 18% of lignin from wheat straw. Substantial lignin removal (37%) was also produced when the enzyme-mediator pretreatment was not combined with the alkaline peroxide extraction. Two-dimensional nuclear magnetic resonance (2D NMR) analysis of the whole pretreated wheat straw material swollen in dimethylsulfoxide-d 6 revealed modifications of the lignin polymer, including the lower number of aliphatic side-chains involved in main β-O-4' and β-5' inter-unit linkages per aromatic lignin unit. Simultaneously, the removal of phydroxyphenyl, guaiacyl and syringyl lignin units and of p-coumaric and ferulic acids, as well as a moderate decrease of tricin units, was observed without a substantial change in the wood polysaccharide signals. Especially noteworthy was the formation of Cαoxidized lignin units during the enzymatic treatment.
Four lignin samples were extracted from sugar cane bagasse using four different alcohols (methanol, ethanol, n-propanol, and 1-butanol) via the organosolv-CO2 supercritical pulping process. Langmuir films were characterized by surface pressure vs mean molecular area (Pi-A) isotherms to exploit information at the molecular level carrying out stability tests, cycles of compression/expansion (hysteresis), subphase temperature variations, and metallic ions dissolved into the water subphase at different concentrations. Briefly, it was observed that these lignins are relatively stable on the water surface when compared to those obtained via different extraction processes. Besides, the Pi-A isotherms are shifted to smaller molecular areas at higher subphase temperatures and to larger molecular areas when the metallic ions are dissolved into the subphase. The results are related to the formation of stable aggregates (domains) onto the water subphase by these lignins, as shown in the Pi-A isotherms. It was found as well that the most stable lignin monolayer onto the water subphase is that extracted with 1-butanol. Homogeneous Langmuir-Blodgett (LB) films of this lignin could be produced as confirmed by UV-vis absorption spectroscopy and the cumulative transfer parameter. In addition, FTIR analysis showed that this lignin LB film is structured in a way that the phenyl groups are organized preferentially parallel to the substrate surface. Further, these LB films were deposited onto gold interdigitated electrodes and ITO and applied in studies involving the detection of Cd+2 ions in aqueous solutions at low concentration levels through impedance spectroscopy and electrochemical measurements. FTIR spectroscopy was carried out before and after soaking the thin films into Cd+2 aqueous solutions, revealing a possible physical interaction between the lignin phenyl groups and the heavy metal ions. The importance of using nanostructured systems is demonstrated as well by comparing both LB and cast films.
Sugarcane bagasse and straw, two major agro-industrial byproducts generated by the sugarcane industry, contain significant amounts of carbohydrates that can be hydrolyzed and then converted into ethanol or other valuable compounds. However, access to them is limited by the presence of lignin, a recalcitrant polymer that protects cell-wall polysaccharides from enzymatic hydrolysis. This work demonstrates the ability of an enzymatic pretreatment, based on the laccase from Pycnoporus cinnabarinus, and 1-hydroxybenzotriazole as mediator, to remove and/or modify lignin in sugarcane bagasse and straw residues, improving their subsequent saccharification. Up to 27% and 31% decreases of relative lignin content in ground sugarcane bagasse and straw, respectively, were achieved by the laccase-mediator pretreatment followed by alkaline peroxide extraction. Moreover, the lignin removal directly correlated with improvements in enzymatic saccharification, increasing glucose releases by around 39% and 46% for bagasse and straw, respectively, compared with those of the corresponding controls. Lignin depolymerization and degradation were made evident in the 2D-NMR spectra by a significant reduction in the number of aliphatic side chains involved in the main β-O-4′ and β-5′ interunit linkages, together with a remarkable removal of p-hydroxyphenyl (H), guaiacyl (G), and syringyl (S) lignin units as well as the associated p-coumarates and ferulates, with respect to polysaccharides.
RESUMO-Objetivou-se determinar o efeito de níveis de lisina digestível, utilizando-se o conceito de proteína ideal, sobre o desempenho e as características de carcaça de 40 leitões machos castrados (peso inicial de 30,02 + 1,38 kg e peso final 60,44 + 1,81 kg) de alto potencial genético para deposição de carne magra. Os animais foram distribuídos em delineamento de blocos ao acaso, com quatro tratamentos, cinco repetições e dois animais por unidade experimental. Os tratamentos consistiram de uma ração basal (18,08% de PB e 3.250 kcal de EM/kg) suplementada com L-lisina HCL, resultando em rações com 0,80; 0,90; 1,00 e 1,10% de lisina digestível. As rações foram suplementadas com níveis crescentes de aminoácidos sintéticos, mantendo-se constantes as relações entre os níveis desses aminoácidos essenciais e os de lisina, com base na digestibilidade verdadeira. Não houve efeito dos tratamentos sobre o consumo de ração, o ganho de peso diário e a concentração de uréia no plasma dos animais. Observou-se comportamento linear da conversão alimentar, que melhorou, e do consumo de lisina digestível diário, que aumentou em função dos níveis de lisina digestível da ração. Os tratamentos não influenciaram as porcentagens de água, proteína e gordura e a deposição de gordura na carcaça dos animais. Os níveis de lisina digestível afetaram de forma linear crescente a deposição de proteína na carcaça. O nível de 1,10% de lisina digestível, correspondente a um consumo de lisina digestível de 21,94 g/dia (3,43 g de Lis/Mcal de EM) proporcionou os melhores resultados de desempenho e características de carcaça de suínos machos castrados de alto potencial genético dos 30 aos 60 kg. Palavras-chave: aminoácidos, carcaça, fase de crescimento, genótipo, uréia Dietary digestible lysine requirements, based on the ideal protein concept, for barrows with high genetic potential from 30 to 60 kg ABSTRACT-The effect of increasing dietary digestible lysine levels, based on the ideal protein concept, on performance and carcass characteristics of forty barrows with high genetic potential for lean gain averaging initial and final body weights of 30.02 + 1.38 kg and 60.44 + 1.81 kg, respectively, was evaluated in this experiment. The animals were assigned to a randomized blocks design, with four treatments, five replicates and two animals per experimental unit. The treatments consisted of a basal diet (18.08% of CP and 3,250 kcal of ME/kg) supplemented with increasing levels of HCL-L-lysine (0.80, 0.90, 1.00, and 1.10% of digestible lysine). The diets were supplemented with constant synthetic amino acids:lysine ratios based on the true digestibility. No treatment effects on feed intake, daily weight gain and plasma urea concentration were observed. Feed:gain ratio and digestible lysine intake increased linearly as the dietary digestible lysine levels increased. No significant treatment effects on percentages of water, protein and fat and carcass fat deposition were observed. It was observed increasing linear effect of treatments on carcass protein dep...
Lignins were isolated from spruce, wheat straw, and eucalyptus by using the milled wood lignin (MWL) method. Functional groups and compositional analyses were assessed via 2D NMR and P NMR to realize their effect on enzyme binding. Films of the lignins were fabricated and ellipsometry, atomic force microscopy, and water contact angle measurements were used for their characterization and to reveal the changes upon enzyme adsorption. Moreover, lignin thin films were deposited on quartz crystal microgravimetry (QCM) and surface plasmon (SPR) resonance sensors and used to gain further insights into the lignin-cellulase interactions. For this purpose, a commercial multicomponent enzyme system and a monocomponent Trichoderma reesei exoglucanase (CBH-I) were considered. Strong enzyme adsorption was observed on the various lignins but compared to the multicomponent cellulases, CBH-I displayed lower surface affinity and higher binding reversibility. This resolved prevalent questions related to the affinity of this enzyme with lignin. Remarkably, a strong correlation between enzyme binding and the syringyl/guaiacyl (S/G) ratio was found for the lignins, which presented a similar hydroxyl group content (P NMR): higher protein affinity was determined on isolated spruce lignin (99% G units), while the lowest adsorption occurred on isolated eucalyptus lignin (70% S units). The effect of electrostatic interactions in enzyme adsorption was investigated by SPR, which clearly indicated that the screening of charges allowed more extensive protein adsorption. Overall, this work furthers our understanding of lignin-cellulase interactions relevant to biomass that has been subjected to no or little pretreatment and highlights the widely contrasting effects of the nature of lignin, which gives guidance to improve lignocellulosic saccharification and related processes.
RESUMO -Objetivou-se determinar o efeito de níveis de lisina digestível, utilizando-se o conceito de proteína ideal, sobre o desempenho e as características de carcaça de 40 leitões machos castrados (peso inicial de 30,02 + 1,38 kg e peso final 60,44 + 1,81 kg) de alto potencial genético para deposição de carne magra. Os animais foram distribuídos em delineamento de blocos ao acaso, com quatro tratamentos, cinco repetições e dois animais por unidade experimental. Os tratamentos consistiram de uma ração basal (18,08% de PB e 3.250 kcal de EM/kg) suplementada com L-lisina HCL, resultando em rações com 0,80; 0,90; 1,00 e 1,10% de lisina digestível. As rações foram suplementadas com níveis crescentes de aminoácidos sintéticos, mantendo-se constantes as relações entre os níveis desses aminoácidos essenciais e os de lisina, com base na digestibilidade verdadeira. Não houve efeito dos tratamentos sobre o consumo de ração, o ganho de peso diário e a concentração de uréia no plasma dos animais.Observou-se comportamento linear da conversão alimentar, que melhorou, e do consumo de lisina digestível diário, que aumentou em função dos níveis de lisina digestível da ração. Os tratamentos não influenciaram as porcentagens de água, proteína e gordura e a deposição de gordura na carcaça dos animais. Os níveis de lisina digestível afetaram de forma linear crescente a deposição de proteína na carcaça. O nível de 1,10% de lisina digestível, correspondente a um consumo de lisina digestível de 21,94 g/dia (3,43 g de Lis/Mcal de EM) proporcionou os melhores resultados de desempenho e características de carcaça de suínos machos castrados de alto potencial genético dos 30 aos 60 kg.Palavras-chave: aminoácidos, carcaça, fase de crescimento, genótipo, uréia Dietary digestible lysine requirements, based on the ideal protein concept, for barrows with high genetic potential from 30 to 60 kg ABSTRACT -The effect of increasing dietary digestible lysine levels, based on the ideal protein concept, on performance and carcass characteristics of forty barrows with high genetic potential for lean gain averaging initial and final body weights of 30.02 + 1.38 kg and 60.44 + 1.81 kg, respectively, was evaluated in this experiment. The animals were assigned to a randomized blocks design, with four treatments, five replicates and two animals per experimental unit. The treatments consisted of a basal diet (18.08% of CP and 3,250 kcal of ME/kg) supplemented with increasing levels of HCL-L-lysine (0.80, 0.90, 1.00, and 1.10% of digestible lysine). The diets were supplemented with constant synthetic amino acids:lysine ratios based on the true digestibility. No treatment effects on feed intake, daily weight gain and plasma urea concentration were observed. Feed:gain ratio and digestible lysine intake increased linearly as the dietary digestible lysine levels increased. No significant treatment effects on percentages of water, protein and fat and carcass fat deposition were observed. It was observed increasing linear effect of treatments on carcass protein dep...
Este trabalho foi conduzido com o objetivo de verificar o efeito da adição de antibiótico, probiótico, prebiótico e gluconato de sódio para suínos no período de 28 a 142 dias de idade, correspondente às fases de creche, crescimento e terminação. Foram utilizados 168 leitões desmamados aos 21 dias de idade, distribuídos em delineamento inteiramente casualizado em 42 baias com quatro suínos machos castrados em cada unidade experimental, com seis repetições e sete dietas: ração basal (controle negativo); ração basal + antibiótico; ração basal + probiótico; ração basal + prebiótico; ração basal + probiótico + prebiótico (simbiótico); ração basal + gluconato de sódio; ração basal + gluconato de sódio + probiótico. O desempenho dos animais diferiu entre as dietas e foi melhor nos animais que receberam ração com aditivos no período de 28 a 142 dias de idade. Entre os aditivos, o simbiótico foi mais eficiente, pois promoveu os melhores resultados de ganho de peso e conversão alimentar. Do mesmo modo, a adição de gluconato de sódio + probiótico, prebiótico e probiótico nas rações foi mais eficiente que a adição de antibiótico. A utilização dos aditivos avaliados em substituição ao antibiótico não compromete as características de carcaça e a morfometria intestinal. A associação do probiótico e prebiótico (simbiótico) estudados melhora o desempenho dos suínos nas fases de creche, crescimento e terminação.
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