Effect of aqueous extracts from Ceriporiopsis subvermispora-biotreated wood on the decolorization of Azure B by Fenton-like reactions

Aqueous extracts from wood biotreated with the white-rot fungus Ceriporiopsis subvermispora were evaluated for their Fe3+- and Cu2+-reducing activities and their anti- or prooxidant properties in Fenton-like reactions to decolorize the recalcitrant dye Azure B. The decolorization of Azure B was strongly inhibited in the presence of 10% (v/v) wood extracts. Only 0.1% (v/v)-diluted extracts provided some enhancement of the Azure B decolorization. The iron-containing reactions decolorized more Azure B and consumed substantially more H2O2 than the reactions containing copper. This study demonstrates that water-soluble wood phenols exert anti- or prooxidant effects that depend on their concentration in the reactions and on the type of cation, Fe3+ or Cu2+, used to convert H2O2 to OH radicals. Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.

Effects of exogenous calcium or oxalic acid on Pinus taeda treatment with the white-rot fungus Ceriporiopsis subvermispora

Pinus taeda wood chips were treated with the biopulping fungus Ceriporiopsis subvermispora in calcium-or oxalic acid-amended cultures. The secretion of hydrolytic and oxidative enzymes was inhibited only in the cultures having the highest concentration of calcium (1400 mg kg(-1) wood). Calcium decreased the availability of free oxalic acid, inhibited fungal growth, and reduced lignin mineralization and transformations. Oxalic acid amendment in the cultures was found not to affect the lignin mineralization and transformations; however, it did inhibit the depolymerization reactions detectable in the residual lignin that was retained in the biotreated wood. C. subvermispora presented catabolic activity for oxalic acid in the cultures amended with 1660 mg acid kg(-1) wood, whereas oxalic acid was synthesized when it was amended at low amounts or initially absent in the cultures. These data suggest one ideal ratio of oxalic acid in C. subvermispora cultures and indicate that its exogenous addition does not necessarily accompany the further degradation of lignin. Crown Copyright (C) 2012 Published by Elsevier Ltd. All rights reserved.

Qualidade de sementes de soja colhidas de forma manual e mecânica com diferentes teores de água

A colheita é uma das principais etapas no processo de produção de sementes de soja. O momento adequado para se efetuar a colheita pode variar em função do tipo de colheita e do grau de umidade das sementes. Neste sentido, o objetivo dessa pesquisa foi determinar as alterações fisiológicas de sementes de soja colhidas de forma manual e mecânica com diferentes teores de água. Para tanto, utilizaram-se sementes das cultivares Embrapa 48 e FTS Águia, que têm diferenças no teor de lignina do tegumento, colhidas de forma manual e mecanicamente com 18,0; 15,0 e 12,0% de água. Após a colheita, as sementes foram submetidas à secagem, e a qualidade das sementes foi avaliada pelos testes de germinação, emergência de plântulas, tetrazólio (viabilidade e vigor), envelhecimento acelerado e índice de velocidade de emergência de plântula. Em seguida, as sementes foram armazenadas (20 °C e 45% UR do ar) e analisadas logo após a colheita e aos seis meses de armazenamento. Os resultados obtidos indicam que o parâmetro fisiológico das sementes de soja, não é afetado, quando: se realiza colheita manual com grau de umidade das sementes de 11,4% a 18,4%; e para sementes colhidas à máquina, com grau de umidade entre 12,0% a 15,9%; mantendo a qualidade fisiológica mesmo após seis meses de armazenamento.

Composites from a forest biorefinery byproduct and agrofibers: Lignosulfonate-phenolic type matrices reinforced with sisal fibers

The replacement of phenol with sodium lignosulfonate and formaldehyde with glutaraldehyde in the preparation of resins resulted in a new resol-type phenolic resin, sodium lignosulfonate-glutaraldehyde resin, in addition to sodium lignosulfonate-formaldehyde and phenol-formaldehyde resins. These resins were then used to prepare thermosets and composites reinforced with sisal fibers. Different techniques were used to characterize raw materials and/or thermosets and composites, including inverse gas chromatography, thermogravimetric analysis, and mechanical impact and flexural tests. The substitution of phenol by sodium lignosulfonate in the formulation of the composite matrices increased the impact strength of the respective composites from approximately 400 Jm(-1) to 800 J m(-1) and 1000 J m(-1), showing a considerable enhancement from the replacement of phenol with sodium lignosulfonate. The wettability of the sisal fibers increased when the resins were prepared from sodium lignosulfonate, generating composites in which the adhesion at the fiber-matrix interface was stronger and favored the transference of load from the matrix to the fiber during impact. Results suggested that the composites experienced a different mechanism of load transfer from the matrix to the fiber when a bending load was applied, compared to that experienced during impact. The thermogravimetric analysis results demonstrated that the thermal stability of the composites was not affected by the use of sodium lignosulfonate as a phenolic-type reagent during the preparation of the matrices.

LC-DAD/ESI-MS/MS study of phenolic compounds in ash (Fraxinus excelsior L. and F. americana L.) heartwood. Effect of toasting intensity at cooperage

The phenolic composition of heartwood extracts from Fraxinus excelsior L. and F. americana L., both before and after toasting in cooperage, was studied using LC-DAD/ESI-MS/MS. Low-molecular weight (LMW) phenolic compounds, secoiridoids, phenylethanoid glycosides, dilignols and oligolignols compounds were detected, and 48 were identified, or tentatively characterized, on the basis of their retention time, UV/Vis and MS spectra, and MS fragmentation patterns. Some LMW phenolic compounds like protocatechuic acid and aldehyde, hydroxytyrosol and tyrosol, were unlike to those for oak wood, while ellagic and gallic acid were not found. The toasting of wood resulted in a progressive increase in lignin degradation products with regard to toasting intensity. The levels of some of these compounds in medium-toasted ash woods were much higher than those normally detected in toasted oak, highlighting vanillin levels, thus a more pronounced vanilla character can be expected when using toasted ash wood in the aging wines. Moreover, in seasoned wood, we found a great variety of phenolic compounds which had not been found in oak wood, especially oleuropein, ligstroside and olivil, along with verbascoside and isoverbascoside in F. excelsior, and oleoside in F. americana. Toasting mainly provoked their degradation, thus in medium-toasted wood, only four of them were detected. This resulted in a minor differentiation between toasted ash and oak woods. The absence of tannins in ash wood, which are very important in oak wood, is another peculiar characteristic that should be taken into account when considering its use in cooperage. Copyright (C) 2012 John Wiley & Sons, Ltd.

Unraveling the structure of sugarcane bagasse after soaking in concentrated aqueous ammonia (SCAA) and ethanol production by Scheffersomyces (Pichia) stipitis

Abstract Background Fuel ethanol production from sustainable and largely abundant agro-residues such as sugarcane bagasse (SB) provides long term, geopolitical and strategic benefits. Pretreatment of SB is an inevitable process for improved saccharification of cell wall carbohydrates. Recently, ammonium hydroxide-based pretreatment technologies have gained significance as an effective and economical pretreatment strategy. We hypothesized that soaking in concentrated aqueous ammonia-mediated thermochemical pretreatment (SCAA) would overcome the native recalcitrance of SB by enhancing cellulase accessibility of the embedded holocellulosic microfibrils. Results In this study, we designed an experiment considering response surface methodology (Taguchi method, L8 orthogonal array) to optimize sugar recovery from ammonia pretreated sugarcane bagasse (SB) by using the method of soaking in concentrated aqueous ammonia (SCAA-SB). Three independent variables: ammonia concentration, temperature and time, were selected at two levels with center point. The ammonia pretreated bagasse (SCAA-SB) was enzymatically hydrolysed by commercial enzymes (Celluclast 1.5 L and Novozym 188) using 15 FPU/g dry biomass and 17.5 Units of β-glucosidase/g dry biomass at 50°C, 150 rpm for 96 h. A maximum of 28.43 g/l reducing sugars corresponding to 0.57 g sugars/g pretreated bagasse was obtained from the SCAA-SB derived using a 20% v/v ammonia solution, at 70°C for 24 h after enzymatic hydrolysis. Among the tested parameters, pretreatment time showed the maximum influence (p value, 0.053282) while ammonia concentration showed the least influence (p value, 0.612552) on sugar recovery. The changes in the ultra-structure and crystallinity of native SCAA-SB and enzymatically hydrolysed SB were observed by scanning electron microscopy (SEM), x-ray diffraction (XRD) and solid-state 13C nuclear magnetic resonance (NMR) spectroscopy. The enzymatic hydrolysates and solid SCAA-SB were subjected to ethanol fermentation under separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) by Scheffersomyces (Pichia) stipitis NRRL Y-7124 respectively. Higher ethanol production (10.31 g/l and yield, 0.387 g/g) was obtained through SSF than SHF (3.83 g/l and yield, 0.289 g/g). Conclusions SCAA treatment showed marked lignin removal from SB thus improving the accessibility of cellulases towards holocellulose substrate as evidenced by efficient sugar release. The ultrastructure of SB after SCAA and enzymatic hydrolysis of holocellulose provided insights of the degradation process at the molecular level.

Chemical and morphological characterization of sugarcane bagasse submitted to a delignification process for enhanced enzymatic digestibility

Abstract Background In recent years, biorefining of lignocellulosic biomass to produce multi-products such as ethanol and other biomaterials has become a dynamic research area. Pretreatment technologies that fractionate sugarcane bagasse are essential for the successful use of this feedstock in ethanol production. In this paper, we investigate modifications in the morphology and chemical composition of sugarcane bagasse submitted to a two-step treatment, using diluted acid followed by a delignification process with increasing sodium hydroxide concentrations. Detailed chemical and morphological characterization of the samples after each pretreatment condition, studied by high performance liquid chromatography, solid-state nuclear magnetic resonance, diffuse reflectance Fourier transformed infrared spectroscopy and scanning electron microscopy, is reported, together with sample crystallinity and enzymatic digestibility. Results Chemical composition analysis performed on samples obtained after different pretreatment conditions showed that up to 96% and 85% of hemicellulose and lignin fractions, respectively, were removed by this two-step method when sodium hydroxide concentrations of 1% (m/v) or higher were used. The efficient lignin removal resulted in an enhanced hydrolysis yield reaching values around 100%. Considering the cellulose loss due to the pretreatment (maximum of 30%, depending on the process), the total cellulose conversion increases significantly from 22.0% (value for the untreated bagasse) to 72.4%. The delignification process, with consequent increase in the cellulose to lignin ratio, is also clearly observed by nuclear magnetic resonance and diffuse reflectance Fourier transformed infrared spectroscopy experiments. We also demonstrated that the morphological changes contributing to this remarkable improvement occur as a consequence of lignin removal from the sample. Bagasse unstructuring is favored by the loss of cohesion between neighboring cell walls, as well as by changes in the inner cell wall structure, such as damaging, hole formation and loss of mechanical resistance, facilitating liquid and enzyme access to crystalline cellulose. Conclusions The results presented herewith show the efficiency of the proposed method for improving the enzymatic digestibility of sugarcane bagasse and provide understanding of the pretreatment action mechanism. Combining the different techniques applied in this work warranted thorough information about the undergoing morphological and chemical changes and was an efficient approach to understand the morphological effects resulting from sample delignification and its influence on the enhanced hydrolysis results.

Treatment of rice straw hemicellulosic hydrolysates with advanced oxidative processes: a new and promising detoxification method to improve the bioconversion process

Abstract Background The use of lignocellulosic constituents in biotechnological processes requires a selective separation of the main fractions (cellulose, hemicellulose and lignin). During diluted acid hydrolysis for hemicellulose extraction, several toxic compounds are formed by the degradation of sugars and lignin, which have ability to inhibit microbial metabolism. Thus, the use of a detoxification step represents an important aspect to be considered for the improvement of fermentation processes from hydrolysates. In this paper, we evaluated the application of Advanced Oxidative Processes (AOPs) for the detoxification of rice straw hemicellulosic hydrolysate with the goal of improving ethanol bioproduction by Pichia stipitis yeast. Aiming to reduce the toxicity of the hemicellulosic hydrolysate, different treatment conditions were analyzed. The treatments were carried out according to a Taguchi L16 orthogonal array to evaluate the influence of Fe+2, H2O2, UV, O3 and pH on the concentration of aromatic compounds and the fermentative process. Results The results showed that the AOPs were able to remove aromatic compounds (furan and phenolic compounds derived from lignin) without affecting the sugar concentration in the hydrolysate. Ozonation in alkaline medium (pH 8) in the presence of H2O2 (treatment A3) or UV radiation (treatment A5) were the most effective for hydrolysate detoxification and had a positive effect on increasing the yeast fermentability of rice straw hemicellulose hydrolysate. Under these conditions, the higher removal of total phenols (above 40%), low molecular weight phenolic compounds (above 95%) and furans (above 52%) were observed. In addition, the ethanol volumetric productivity by P. stipitis was increased in approximately twice in relation the untreated hydrolysate. Conclusion These results demonstrate that AOPs are a promising methods to reduce toxicity and improve the fermentability of lignocellulosic hydrolysates.

Effects of pretreatment on morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production – part 1

Abstract Background In recent years, the growing demand for biofuels has encouraged the search for different sources of underutilized lignocellulosic feedstocks that are available in sufficient abundance to be used for sustainable biofuel production. Much attention has been focused on biomass from grass. However, large amounts of timber residues such as eucalyptus bark are available and represent a potential source for conversion to bioethanol. In the present paper, we investigate the effects of a delignification process with increasing sodium hydroxide concentrations, preceded or not by diluted acid, on the bark of two eucalyptus clones: Eucalyptus grandis (EG) and the hybrid, E. grandis x urophylla (HGU). The enzymatic digestibility and total cellulose conversion were measured, along with the effect on the composition of the solid and the liquor fractions. Barks were also assessed using Fourier-transform infrared spectroscopy (FTIR), solid-state nuclear magnetic resonance (NMR), X-Ray diffraction, and scanning electron microscopy (SEM). Results Compositional analysis revealed an increase in the cellulose content, reaching around 81% and 76% of glucose for HGU and EG, respectively, using a two-step treatment with HCl 1%, followed by 4% NaOH. Lignin removal was 84% (HGU) and 79% (EG), while the hemicellulose removal was 95% and 97% for HGU and EG, respectively. However, when we applied a one-step treatment, with 4% NaOH, higher hydrolysis efficiencies were found after 48 h for both clones, reaching almost 100% for HGU and 80% for EG, in spite of the lower lignin and hemicellulose removal. Total cellulose conversion increased from 5% and 7% to around 65% for HGU and 59% for EG. NMR and FTIR provided important insight into the lignin and hemicellulose removal and SEM studies shed light on the cell-wall unstructuring after pretreatment and lignin migration and precipitation on the fibers surface, which explain the different hydrolysis rates found for the clones. Conclusion Our results show that the single step alkaline pretreatment improves the enzymatic digestibility of Eucalyptus bark. Furthermore, the chemical and physical methods combined in this study provide a better comprehension of the pretreatment effects on cell-wall and the factors that influence enzymatic digestibility of this forest residue.

Uso de aditivos na biodegradação de madeira pelo fungo Ceriporiopsis subvermispora: efeito na peroxidação de lipídios dependente de manganês-peroxidase

Ceriporiopsis subvermispora is a selective fungus in the wood delignification and the most promising in biopulping. Through the lipid peroxidation initiated by manganese peroxidase (MnP), free radicals can be generated, which can act in the degradation of lignin nonphenolic structures. This work evaluated the prooxidant activity (based in lipid peroxidation) of enzymatic extracts from wood biodegradation by this fungus in cultures containing exogenous calcium, oxalic acid or soybean oil. It was observed that MnP significant activity is required to promote lipid peroxidation and wood delignification. Positive correlation between prooxidant activity x MnP was observed up to 300 IU kg-1 of wood.

Teores de lignina e celulose em plantas de cana-de-açúcar em função da aplicação de maturadores

O objetivo deste trabalho foi avaliar os teores de lignina e celulose em plantas de cana-de-açúcar após a aplicação de dois maturadores para a colheita. O experimento foi conduzido em uma área de cana-soca, cultivar SP 803280, no município de Igaraçu do Tietê/SP. O delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. Os tratamentos constituíram-se da aplicação de dois maturadores: sulfometuron-methyl (Curavial) e glyphosate (Roundup original). As doses utilizadas foram: glyphosate a 72 g e.a. ha-1; glyphosate a 144 g e.a. ha-1 ; glyphosate a 72 g e.a. ha-1 + sulfometuron methyl a 10 g p.c. ha-1; glyphosate a 108 g e.a. ha-1 + sulfometuron-methyl a 12 g p.c. ha-1; sulfometuron-methyl a 20 g p.c. ha-1; e a testemunha sem aplicação de maturadores. As análises de lignina e celulose foram realizadas pelo método lignina em detergente ácido modificado. O glyphosate e o sulfometuron-methyl alteraram os níveis de lignina no momento da colheita, e esse efeito foi observado também durante o crescimento da cana-de-açúcar (meses após a aplicação desses produtos). O glyphosate a 72 g e.a. ha-1 promoveu reduções nos teores de lignina, na colheita e durante o crescimento da cana-de-açúcar, quando comparados com os da testemunha, enquanto o sulfometuron-methyl isolado na menor dose (10 g ha-1) promoveu aumento nos teores desse biopolímero na soqueira da cana-de-açúcar.

Effect of thermal treatment on the chemical characteristics of wood from Eucalyptus grandis W. Hill ex Maiden under different atmospheric conditions

Thermal treatment (thermal rectification) is a process in which technological properties of wood are modified using thermal energy, the result of which is often value-added wood. Thermally treated wood takes on similar color shades to tropical woods and offers considerable resistance to destructive microorganisms and climate action, in addition to having high dimensional stability and low hygroscopicity. Wood samples of Eucalyptus grandis were subjected to various thermal treatments, as performed in presence (140ºC; 160ºC; 180ºC) or in absence of oxygen (160ºC; 180ºC; 200ºC) inside a thermal treatment chamber, and then studied as to their chemical characteristics. Increasing the maximum treatment temperatures led to a reduction in the holocellulose content of samples as a result of the degradation and volatilization of hemicelluloses, also leading to an increase in the relative lignin content. Except for glucose, all monosaccharide levels were found to decrease in samples after the thermal treatment at a maximum temperature of 200ºC. The thermal treatment above 160ºC led to increased levels of total extractives in the wood samples, probably ascribed to the emergence of low molecular weight substances as a result of thermal degradation. Overall, it was not possible to clearly determine the effect of presence or absence of oxygen in the air during thermal treatment on the chemical characteristics of the relevant wood samples.

Germination, carbohydrate composition and vigor of cryopreserved Caesalpinia echinata seeds

The present study investigated the germination and vigor of Caesalpinia echinata (Brazilwood) seeds stored at negative temperatures. Recently harvested seeds were cryopreserved at -18º or -196ºC and periodically evaluated for germination, seed vigor and carbohydrate composition. The temperatures did not influence the germination percentages or vigor. The germination percentage decreased from 88% in recently harvested seeds to 60% after 730 days of storage. The different temperature and storage times tested did not affect the vigor seed germination as indicated by the measures of plant growth and survival. The different temperatures used did not cause changes in the carbohydrate composition. The tegument cell walls were rich in lignin, arabinose and xylose. The cytoplasm of the cotyledons and embryos had high levels of glucose, fructose, and sucrose. The cryopreservation technique here presented was effective in the conservation of Brazilwood seeds for the medium term.

Bromatological characteristics and in vitro digestibility of four sugarcane varieties subjected or not to the application of quicklime

The experiment was conducted to evaluate the bromatological characteristics and the in vitro digestibility of four sugarcane varieties, subjected or not to hydrolysis, with quicklime. A completely randomized design was employed with three replications arranged in a 4 × 2 factorial scheme, with four sugarcane varieties (SP 52454, RB 867515, RB 855536 and IAC 862480), hydrolyzed or not. There was significant effect on brix (p < 0.05) and industrial fiber (p < 0.05), and IAC 862480 variety had the lowest levels of industrial fiber. There were no significant difference (p > 0.05) in neutral detergent fiber, acid detergent fiber and lignin levels among the sugarcane varieties under analysis and for the sugarcanes, hydrolyzed or not. The use of sugarcane hydrolysis with 1% quicklime improves the in vitro digestibility of NDF and ADF, regardless of the variety studied. Hydrolysis with 1% quicklime did not alter the sugarcane chemical composition.

Environmental assessment of residues generated after consecutive acid-base pretreatment of sugarcane bagasse by advanced oxidative process

Abstract Background Biofuels produced from sugarcane bagasse (SB) have shown promising results as a suitable alternative of gasoline. Biofuels provide unique, strategic, environmental and socio-economic benefits. However, production of biofuels from SB has negative impact on environment due to the use of harsh chemicals during pretreatment. Consecutive sulfuric acid-sodium hydroxide pretreatment of SB is an effective process which eventually ameliorates the accessibility of cellulase towards cellulose for the sugars production. Alkaline hydrolysate of SB is black liquor containing high amount of dissolved lignin. Results This work evaluates the environmental impact of residues generated during the consecutive acid-base pretreatment of SB. Advanced oxidative process (AOP) was used based on photo-Fenton reaction mechanism (Fenton Reagent/UV). Experiments were performed in batch mode following factorial design L9 (Taguchi orthogonal array design of experiments), considering the three operation variables: temperature (°C), pH, Fenton Reagent (Fe2+/H2O2) + ultraviolet. Reduction of total phenolics (TP) and total organic carbon (TOC) were responsive variables. Among the tested conditions, experiment 7 (temperature, 35°C; pH, 2.5; Fenton reagent, 144 ml H2O2+153 ml Fe2+; UV, 16W) revealed the maximum reduction in TP (98.65%) and TOC (95.73%). Parameters such as chemical oxygen demand (COD), biochemical oxygen demand (BOD), BOD/COD ratio, color intensity and turbidity also showed a significant change in AOP mediated lignin solution than the native alkaline hydrolysate. Conclusion AOP based on Fenton Reagent/UV reaction mechanism showed efficient removal of TP and TOC from sugarcane bagasse alkaline hydrolysate (lignin solution). To the best of our knowledge, this is the first report on statistical optimization of the removal of TP and TOC from sugarcane bagasse alkaline hydrolysate employing Fenton reagent mediated AOP process.