Adriane M. F. Milagres

A study on the pretreatment of a sugarcane bagasse sample with dilute sulfuric acid.

Experiments based on a 23 central composite full factorial design were carried out in 200-ml stainless-steel containers to study the pretreatment, with dilute sulfuric acid, of a sugarcane bagasse sample obtained from a local sugar–alcohol mill. The independent variables selected for study were temperature, varied from 112.5°C to 157.5°C, residence time, varied from 5.0 to 35.0 min, and sulfuric acid concentration, varied from 0.0% to 3.0% (w/v). Bagasse loading of 15% (w/w) was used in all experiments. Statistical analysis of the experimental results showed that all three independent variables significantly influenced the response variables, namely the bagasse solubilization, efficiency of xylose recovery in the hemicellulosic hydrolysate, efficiency of cellulose enzymatic saccharification, and percentages of cellulose, hemicellulose, and lignin in the pretreated solids. Temperature was the factor that influenced the response variables the most, followed by acid concentration and residence time, in that order. Although harsher pretreatment conditions promoted almost complete removal of the hemicellulosic fraction, the amount of xylose recovered in the hemicellulosic hydrolysate did not exceed 61.8% of the maximum theoretical value. Cellulose enzymatic saccharification was favored by more efficient removal of hemicellulose during the pretreatment. However, detoxification of the hemicellulosic hydrolysate was necessary for better bioconversion of the sugars to ethanol.

Characterization of xylanase production by a local isolate of Penicillium janthinellum

Abstract A strain of P. janthinellum was isolated locally from plant material found in a termite colony. Comparison with T. reesei revealed high xylanolytic activity. This activity was inducible by xylan, sugar cane bagasse, or xylose. Induction by xylan was repressed by the presence of xylose or glucose. A temperature of 40°C and a pH of 5.5 were optimal conditions for xylanase activity (98 units ml −1 ). Unlike many filamentous fungi, only very low or no endocellulolytic activity could be detected in the presence of various potential inducers, including xylan and cellulose.

Enzymatic hydrolysis of chemithermomechanically pretreated sugarcane bagasse and samples with reduced initial lignin content

Chemithermomechanical (CTM) processing was used to pretreat sugarcane bagasse with the aim of increasing cell wall accessibility to hydrolytic enzymes. Yields of the pretreated samples were in the range of 75–94%. Disk refining and alkaline‐CTM and alkaline/sulfite‐CTM pretreatments yielded pretreated materials with 21.7, 17.8, and 15.3% of lignin, respectively. Hemicellulose content was also decreased to some extent. Fibers of the pretreated materials presented some external fibrillation, fiber curling, increased swelling, and high water retention capacity. Cellulose conversion of the alkaline‐CTM‐ and alkaline/sulfite‐CTM‐pretreated samples reached 50 and 85%, respectively, after 96 h of enzymatic hydrolysis. Two samples with low initial lignin content were also evaluated after the mildest alkaline‐CTM pretreatment. One sample was a partially delignified mill‐processed bagasse. The other was a sugarcane hybrid selected in a breeding program. Samples with lower initial lignin content were hydrolyzed considerably faster in the first 24 h of enzymatic digestion. For example, enzymatic hydrolysis of the sample with the lowest initial lignin content (14.2%) reached 64% cellulose conversion after only 24 h of hydrolysis when compared with the 30% observed for the mill‐processed bagasse containing an initial lignin content of 24.4%.

Uma visão sobre a estrutura, composição e biodegradação da madeira

Wood is the main raw material used in the pulp and paper industry. It is a material that presents heterogeneous structure and complex composition, which results in a relatively resistant material to the biodegradation process. In the present review, we attempted to summarize the structural characteristics of wood and describe the chemical nature of its major components to, afterwards, comment about its biodegradation. The role of the enzyme manganese peroxidase in the lignin degradation by a selective white-rot fungus, Ceriporiopsis subvermispora, was highlighted.

The effects of lignin removal and drying on the porosity and enzymatic hydrolysis of sugarcane bagasse

The porosity of lignocellulosic materials is a key feature for the enzymatic hydrolysis of the constituent polysaccharides, being affected by its drying and lignin content. Here we evaluated the influence of both parameters in the porosity of sugarcane bagasse, using raw and chlorite-delignified samples. A fraction of these samples was air dried at room temperature and the remainder one was kept wet. All the samples were subjected to porometry (solute exclusion technique), determination of water retention value and assessment of enzymatic saccharification of polysaccharides. Samples with higher lignin contents had lower porosities and exhibited worse enzymatic conversions of polysaccharides. Mild drying collapsed only the smaller pores, which already were inaccessible to enzymes, and therefore did not affect the efficiencies of saccharification. Our results show that the lignin content plays an important role in the porosity of the material and that its removal improves the enzymatic saccharification of the constituent polysaccharides.

Production of xylanases from Penicillium janthinellum and its use in the recovery of cellulosic textile fibers

Abstract An isolate of the fungus Penicillium janthinellum that produces large amounts of xylanase with no detectable cellulase activity was used as a model system to study enzyme induction and production and to illustrate the use of such preparations in the recovery of cellulosic textile fibers. For large-scale production of xylanase, the hemicellulosic fraction from steam-exploded sugar cane bagasse (SEBh) was investigated as an abundant and efficient inducer. A two-step cultivation system, in which mycelia were first produced under noninducing conditions and switched to inductive media, demonstrated that xylanase production was dependent on the noninducing carbon source. A consistent decrease in lag periods prior to xylanase detection, as well as a sharp increase of production rates with length of starvation by noninduced cells, suggested that xylanase synthesis only started after intracellular reserves were depleted. These results suggest that plants used to produce textile fibers are potential targets for enzymatic treatments by xylanase-producing organisms.

Direct ethanol production from glucose, xylose and sugarcane bagasse by the corn endophytic fungi Fusarium verticillioides and Acremonium zeae.

Production of ethanol with two corn endophytic fungi, Fusarium verticillioides and Acremonium zeae, was studied. The yield of ethanol from glucose, xylose and a mixture of both sugars were 0.47, 0.46 and 0.50g/g ethanol/sugar for F. verticillioides and 0.37, 0.39 and 0.48g/g ethanol/sugar for A. zeae. Both fungi were able to co-ferment glucose and xylose. Ethanol production from 40g/L of pre-treated sugarcane bagasse was 4.6 and 3.9g/L for F. verticillioides and A. zeae, respectively, yielding 0.31g/g of ethanol per consumed sugar. Both fungi studied were capable of co-fermenting glucose and xylose at high yields. Moreover, they were able to produce ethanol directly from lignocellulosic biomass, demonstrating to be suitable microorganisms for consolidated bioprocessing.

Interference of some aqueous two-phase system phase-forming components in protein determination by the Bradford method.

The interference of some specific aqueous two-phase system (ATPS) phase-forming components in bovine serum albumin (BSA) determination by the Bradford method was investigated. For this purpose, calibration curves were obtained for BSA in the presence of different concentrations of salts and polymers. A total of 19 salts [Na₂SO₄, (NH₄)₂SO₄, MgSO₄, LiSO₄, Na₂HPO₄, sodium phosphate buffer (pH 7.0), NaH₂PO₄, K₂HPO₄, potassium phosphate buffer (pH 7.0), KH₂PO₄, C₆H₈O₇, Na₃C₆H₅O₇, KCHO₂, NaCHO₂, NaCO₃, NaHCO₃, C₂H₄O₂, sodium acetate buffer (pH 4.5), and NaC₂H₃O₂] and 7 polymers [PEG 4000, PEG 8000, PEG 20000, UCON 3900, Ficoll 70000, PES 100000, and PVP 40000] were tested, and each calibration curve was compared with the one obtained for BSA in water. Some concentrations of salts and polymers had considerable effect in the BSA calibration curve. Carbonate salts were responsible for the highest salt interference, whereas citric and acetic acids did not produce interference even in the maximum concentration level tested (5 wt%). Among the polymers, UCON gave the highest interference, whereas Ficoll did not produce interference when used in concentrations up to 10 wt%. It was concluded that a convenient dilution of the samples prior to the protein quantification is needed to ensure no significant interference from ATPS phase-forming constituents.

Biomimetic oxidative treatment of spruce wood studied by pyrolysis-molecular beam mass spectrometry coupled with multivariate analysis and 13C-labeled tetramethylammonium hydroxide thermochemolysis: implications for fungal degradation of wood.

In this work, pyrolysis–molecular beam mass spectrometry analysis coupled with principal components analysis and 13C-labeled tetramethylammonium hydroxide thermochemolysis were used to study lignin oxidation, depolymerization, and demethylation of spruce wood treated by biomimetic oxidative systems. Neat Fenton and chelator-mediated Fenton reaction (CMFR) systems as well as cellulosic enzyme treatments were used to mimic the nonenzymatic process involved in wood brown-rot biodegradation. The results suggest that compared with enzymatic processes, Fenton-based treatment more readily opens the structure of the lignocellulosic matrix, freeing cellulose fibrils from the matrix. The results demonstrate that, under the current treatment conditions, Fenton and CMFR treatment cause limited demethoxylation of lignin in the insoluble wood residue. However, analysis of a water-extractable fraction revealed considerable soluble lignin residue structures that had undergone side chain oxidation as well as demethoxylation upon CMFR treatment. This research has implications for our understanding of nonenzymatic degradation of wood and the diffusion of CMFR agents in the wood cell wall during fungal degradation processes.

Mapping of Cell Wall Components in Lignified Biomass as a Tool to Understand Recalcitrance

Lignocelulosic biomass is recalcitrant to enzymatic digestion because terrestrial plants develop an efficient manner to grow upward and resist the microbial degradation of the polysaccharides contained in their cell walls. The complex cell ultrastructure, varied tissues, and the composite characteristic of the cell walls are among the several factors explaining the recalcitrance of lignified plants. Mapping the macromolecular components in the cell walls has proved to be useful to understand the varied recalcitrance of different biomass tissues. Available data indicate that lignin and hemicellulose greatly affect the final digestibility of the lignocellulosic materials. Removal of these components from the cell walls with varied pretreatments or even using lignin- and/or hemicellulose-depleted plants indicate that a critical characteristic of the cell wall to be digestible is to present most as possible available cellulose. This chapter revises some basic information on cell wall structure and advance in the knowledge compiling information on the mapping of cell wall components by several techniques and showing that the removal of cellulose encapsulating components is a key factor to increase cell wall porosity and digestibility by hydrolytic enzymes.