High yield biorefinery products from sugarcane bagasse: Prebiotic xylooligosaccharides, cellulosic ethanol, cellulose nanofibrils and lignin nanoparticles.

An integrated biorefining strategy was applied to fractionate Sugarcane bagasse (SCB) into its major constituents, enabling high-yield conversion of the fractionated materials into high-value coproducts alongside cellulosic ethanol. Pilot-scale steam explosion produced a hydrolysate rich in low molecular weight xylooligosaccharides that had a high in vitro efficacy as a prebiotic towards different bifidobacteria. Lignin recovered after alkaline treatment of the steam-exploded SCB was converted into uniform spherical lignin nanoparticles (11.3 nm in diameter) by a green mechanical method. The resulting cellulose was hydrolyzed at 17.5% (w/v) consistency and low enzyme loading (17.5 mg/g) to yield a pure glucose hydrolysate at a high concentration (100 g/L) and a cellulosic solid residue that was defibrillated by disc ultra-refining into homogeneous cellulose nanofibrils (20.5 nm in diameter). Statistical optimization of the cellulosic hydrolysate fermentation led to ethanol production of 67.1 g/L, with a conversion yield of 0.48 g/g and productivity of 1.40 g/L.h.

Chemical composition and enzymatic digestibility of sugarcane clones selected for varied lignin content.

BACKGROUND: The recalcitrance of lignocellulosic materials is a major limitation for their conversion into fermentable sugars. Lignin depletion in new cultivars or transgenic plants has been identified as a way to diminish this recalcitrance. In this study, we assessed the success of a sugarcane breeding program in selecting sugarcane plants with low lignin content, and report the chemical composition and agronomic characteristics of eleven experimental hybrids and two reference samples. The enzymatic digestion of untreated and chemically delignified samples was evaluated to advance the performance of the sugarcane residue (bagasse) in cellulosic-ethanol production processes. RESULTS: The ranges for the percentages of glucan, hemicellulose, lignin, and extractive (based on oven-dry biomass) of the experimental hybrids and reference samples were 38% to 43%, 25% to 32%, 17% to 24%, and 1.6% to 7.5%, respectively. The samples with the smallest amounts of lignin did not produce the largest amounts of total polysaccharides. Instead, a variable increase in the mass of a number of components, including extractives, seemed to compensate for the reduction in lignin content. Hydroxycinnamic acids accounted for a significant part of the aromatic compounds in the samples, with p-coumaric acid predominating, whereas ferulic acid was present only in low amounts. Hydroxycinnamic acids with ester linkage to the hemicelluloses varied from 2.3% to 3.6%. The percentage of total hydroxycinnamic acids (including the fraction linked to lignin through ether linkages) varied from 5.0% to 9.2%, and correlated to some extent with the lignin content. These clones released up to 31% of glucose after 72 hours of digestion with commercial cellulases, whereas chemically delignified samples led to cellulose conversion values of more than 80%. However, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment. CONCLUSION: Some of the experimental sugarcane hybrids did have the combined characteristics of high biomass and high sucrose production with low lignin content. Conversion of glucan to glucose by commercial cellulases was increased in the samples with low lignin content. Chemical delignification further increased the cellulose conversion to values of more than 80%. Thus, plants with lower lignin content required less delignification to reach higher efficiencies of cellulose conversion during the enzymatic treatment.

Topochemical distribution of lignin and hydroxycinnamic acids in sugar-cane cell walls and its correlation with the enzymatic hydrolysis of polysaccharides.

BACKGROUND: Lignin and hemicelluloses are the major components limiting enzyme infiltration into cell walls. Determination of the topochemical distribution of lignin and aromatics in sugar cane might provide important data on the recalcitrance of specific cells. We used cellular ultraviolet (UV) microspectrophotometry (UMSP) to topochemically detect lignin and hydroxycinnamic acids in individual fiber, vessel and parenchyma cell walls of untreated and chlorite-treated sugar cane. Internodes, presenting typical vascular bundles and sucrose-storing parenchyma cells, were divided into rind and pith fractions. RESULTS: Vascular bundles were more abundant in the rind, whereas parenchyma cells predominated in the pith region. UV measurements of untreated fiber cell walls gave absorbance spectra typical of grass lignin, with a band at 278 nm and a pronounced shoulder at 315 nm, assigned to the presence of hydroxycinnamic acids linked to lignin and/or to arabino-methylglucurono-xylans. The cell walls of vessels had the highest level of lignification, followed by those of fibers and parenchyma. Pith parenchyma cell walls were characterized by very low absorbance values at 278 nm; however, a distinct peak at 315 nm indicated that pith parenchyma cells are not extensively lignified, but contain significant amounts of hydroxycinnamic acids. Cellular UV image profiles scanned with an absorbance intensity maximum of 278 nm identified the pattern of lignin distribution in the individual cell walls, with the highest concentration occurring in the middle lamella and cell corners. Chlorite treatment caused a rapid removal of hydroxycinnamic acids from parenchyma cell walls, whereas the thicker fiber cell walls were delignified only after a long treatment duration (4 hours). Untreated pith samples were promptly hydrolyzed by cellulases, reaching 63% of cellulose conversion after 72 hours of hydrolysis, whereas untreated rind samples achieved only 20% hydrolyzation. CONCLUSION: The low recalcitrance of pith cells correlated with the low UV-absorbance values seen in parenchyma cells. Chlorite treatment of pith cells did not enhance cellulose conversion. By contrast, application of the same treatment to rind cells led to significant removal of hydroxycinnamic acids and lignin, resulting in marked enhancement of cellulose conversion by cellulases.

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%.

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

Experiments based on a 2(3) 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 commercial cellulases and their use in the saccharification of a sugarcane bagasse sample pretreated with dilute sulfuric acid.

This study aimed to correlate the efficiency of enzymatic hydrolysis of the cellulose contained in a sugarcane bagasse sample pretreated with dilute H(2)SO(4) with the levels of independent variables such as initial content of solids and loadings of enzymes and surfactant (Tween 20), for two cellulolytic commercial preparations. The preparations, designated cellulase I and cellulase II, were characterized regarding the activities of total cellulases, endoglucanase, cellobiohydrolase, cellobiase, ß-glucosidase, xylanase, and phenoloxidases (laccase, manganese and lignin peroxidases), as well as protein contents. Both extracts showed complete cellulolytic complexes and considerable activities of xylanases, without activities of phenoloxidases. For the enzymatic hydrolyses, two 2(3) central composite full factorial designs were employed to evaluate the effects caused by the initial content of solids (1.19-4.81%, w/w) and loadings of enzymes (1.9-38.1 FPU/g bagasse) and Tween 20 (0.0-0.1 g/g bagasse) on the cellulose digestibility. Within 24 h of enzymatic hydrolysis, all three independent variables influenced the conversion of cellulose by cellulase I. Using cellulase II, only enzyme and surfactant loadings showed significant effects on cellulose conversion. An additional experiment demonstrated the possibility of increasing the initial content of solids to values much higher than 4.81% (w/w) without compromising the efficiency of cellulose conversion, consequently improving the glucose concentration in the hydrolysate.

Xylooligosaccharides production from alkali-pretreated sugarcane bagasse using xylanases from Thermoascus aurantiacus.

Sugarcane bagasse hemicellulose was isolated in a one-step chemical extraction using hydrogen peroxide in alkaline media. The polysaccharide containing 80.9% xylose and small amounts of L-arabinose, 4-O-methyl-D-glucuronic acid and glucose, was hydrolyzed by crude enzymatic extracts from Thermoascus aurantiacus at 50 degrees C. Conditions of enzymatic hydrolysis leading to the best yields of xylose and xylooligosaccharides (DP 2-5) were investigated using substrate concentration in the range 0.5-3.5% (w/v), enzyme load 40-80 U/g of the substrate, and reaction time from 3 to 96 h, applying a 2(2) factorial design. The maximum conversion to xylooligosaccharides (37.1%) was obtained with 2.6% of substrate and xylanase load of 60 U/g. The predicted maximum yield of xylobiose by a polynomial model was 41.6%. Crude enzymatic extract of T. aurantiacus generate from sugarcane bagasse hemicellulose 39% of xylose, 59% of xylobiose, and 2% of other xylooligosaccharides.

Ethanol production from sugarcane bagasse hydrolysate using Pichia stipitis.

The objective of this study was to evaluate the ethanol production from the sugars contained in the sugarcane bagasse hemicellulosic hydrolysate with the yeast Pichia stipitis DSM 3651. The fermentations were carried out in 250-mL Erlenmeyers with 100 mL of medium incubated at 200 rpm and 30 degrees C for 120 h. The medium was composed by raw (non-detoxified) hydrolysate or by hydrolysates detoxified by pH alteration followed by active charcoal adsorption or by adsorption into ion-exchange resins, all of them supplemented with yeast extract (3 g/L), malt extract (3 g/L), and peptone (5 g/L). The initial concentration of cells was 3 g/L. According to the results, the detoxification procedures removed inhibitory compounds from the hemicellulosic hydrolysate and, thus, improved the bioconversion of the sugars into ethanol. The fermentation using the non-detoxified hydrolysate led to 4.9 g/L ethanol in 120 h, with a yield of 0.20 g/g and a productivity of 0.04 g L(-1) h(-1). The detoxification by pH alteration and active charcoal adsorption led to 6.1 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.13 g L(-1) h(-1). The detoxification by adsorption into ion-exchange resins, in turn, provided 7.5 g/L ethanol in 48 h, with a yield of 0.30 g/g and a productivity of 0.16 g L(-1) h(-1).

Xylitol production from wheat straw hemicellulosic hydrolysate: hydrolysate detoxification and carbon source used for inoculum preparation / Produção de xilitol em hidrolisado hemicelulósico de palha de trigo: destoxificação do hidrolisado e fonte de carbono utilizada para o preparo do inóculo

Wheat straw hemicellulosic hydrolysate was used for xylitol bioproduction. The use of a xylose-containing medium to grow the inoculum did not favor the production of xylitol in the hydrolysate, which was submitted to a previous detoxification treatment with 2.5 percent activated charcoal for optimized removal of inhibitory compounds.

Hidrolisado hemicelulósico de palha de trigo foi utilizado para a bioprodução de xilitol. O uso de meio contendo xilose para crescer o inóculo não favoreceu a produção de xilitol no hidrolisado, que foi submetido a um tratamento prévio de destoxificação com 2.5 por cento de carvão ativo para remoção otimizada de compostos inibitórios.

Semi-continuous xylose-to-xylitol bioconversion by Ca-alginate entrapped yeast cells in a stirred tank reactor.

Candida guilliermondii FTI 20037 cells were entrapped in Ca-alginate beads and used for xylose-to-xylitol bioconversions during five successive batches in a stirred tank reactor. Supplemented sugarcane bagasse hemicellulosic hydrolysate was used as the fermentation medium. The average volume of the Ca-alginate beads was reduced by about 30% after the 600 h taken to perform the five bioconversion cycles, thus demonstrating physical instability under the conditions prevailing in the reactor vessel. In spite of this, almost steady bioconversion rates and yields were observed along the repeated batches. In average values, a production of 51.6 g l(-1), a productivity of 0.43 g l(-1 )h(-1) and a yield of 0.71 g g(-1) were attained in each batch, variation coefficients being smaller than 10%.

Xylitol production from wheat straw hemicellulosic hydrolysate: hydrolysate detoxification and carbon source used for inoculum preparation.

Wheat straw hemicellulosic hydrolysate was used for xylitol bioproduction. The use of a xylose-containing medium to grow the inoculum did not favor the production of xylitol in the hydrolysate, which was submitted to a previous detoxification treatment with 2.5% activated charcoal for optimized removal of inhibitory compounds.

Semi-continuous xylitol bioproduction in sugarcane bagasse hydrolysate: effect of nutritional supplementation

Xylose-to-xylitol bioconversion by Ca-alginate entrapped Candida guilliermondii cells in sugarcane bagasse hemicellulosic hydrolysate was carried out in erlenmeyer flasks using the repeated-batch mode of fermentation. The hydrolysate was supplemented or not with ammonium sulfate and/or rice bran extract at the beginning of each repeated batch. Altogether, six sets of three repeated-batches were carried out, the immobilized cells being reused at the end of each batch. The best results were achieved when the hydrolysate was supplemented with both nutrients in all the three repeated batches, which provided xylitol productions of 25.9, 46.8, 48.7 gL-1, productivities of 0.27, 0.49, 0.51 gL-1h-1, and yields of 0.45, 0.58, 0.55 gg-1, respectively. In the absence of nutrients, the xylitol production, productivity and yield did not exceed 12.1 gL-1, 0.13 gL-1h-1 and 0.30 gg-1, respectively.

A bioconversão de xilose em xilitol por células de Candida guilliermondii imobilizadas em alginato de cálcio, em hidrolisado hemicelulósico de bagaço de cana-de-açúcar, foi realizada em frascos erlenmeyer no modo bateladas repetidas de fermentação. O hidrolisado foi suplementado ou não com sulfato de amônio e/ou extrato de farelo de arroz no início de cada batelada repetida. No total, seis experimentos com três bateladas repetidas cada um foram realizados, sendo as células imobilizadas reutilizadas ao final de cada batelada. Os melhores resultados foram alcançados quando o hidrolisado foi suplementado com ambos nutrientes em todas as três bateladas repetidas, resultando em concentrações de xilitol iguais a 25,9, 46,8 e 48,7 gL-1, produtividades de 0,27, 0,49 e 0,51 gL-1h-1, e rendimentos de 0,45, 0,58 e 0,55 gg-1, respectivamente. Na ausência de nutrientes, a concentração de xilitol, a produtividade e o rendimento não ultrapassaram 12,1 gL-1, 0,13 gL-1h-1 e 0.30 gg-1, respectivamente.

Effects of sulfuric acid loading and residence time on the composition of sugarcane bagasse hydrolysate and its use as a source of xylose for xylitol bioproduction.

A 2(2) full factorial design was employed to evaluate the effects of sulfuric acid loading and residence time on the composition of sugarcane bagasse hydrolysate obtained in a 250-L reactor. The acid loading and the residence time were varied from 70 to 130 mg acid per gram of dry bagasse and from 10 to 30 min, respectively, while the temperature (121 degrees C) and the bagasse loading (10%) were kept constant. Both the sulfuric acid loading and the residence time influenced the concentrations of xylose and inhibitors in the hydrolysate. The highest xylose concentration (22.71 g/L) was achieved when using an acid loading of 130 mg/g and a residence time of 30 min. These conditions also led to increased concentrations of inhibiting byproducts in the hydrolysate. All of the hydrolysates were vacuum-concentrated to increase the xylose concentration, detoxified by pH alteration and adsorption into activated charcoal, and used for xylitol bioproduction in a stirred tank reactor. Neither the least (70 mg/g, 10 min) nor the most severe (130 mg/g, 30 min) hydrolysis conditions led to the best xylitol production (37.5 g/L), productivity (0.85 g/L h), and yield (0.78 g/g).

Sugarcane bagasse as raw material and immobilization support for xylitol production.

Xylose-to-xylitol bioconversion was performed utilizing Candida guilliermondii immobilized in sugarcane bagasse and cultured in Erlenmeyer flasks using sugarcane bagasse hydrolysate as the source of xylose. Fermentations were carried out according to a factorial design, and the independent variables considered were treatment, average diameter, and amount of bagasse used as support for cell immobilization. By increasing the amount of support, the xylitol yield decreased, whereas the biomass yield increased. The diameter of the support did not influence xylitol production, and treatment of the bagasse with hexamethylene diamine prior to fermentation resulted in the highest amount of immobilized cells.

Hormônio do crescimento ou somatotrófico: novas perspectivas na deficiência isolada de GH a partir da descrição da mutação no gene do receptor do GHRH nos indivíduos da cidade de Itabaianinha, Brasil / Growth or somatotrophic hormone: new perspectives in isolated GH deficiency after description of the mutation in the GHRH receptor gene in individuals of Itabaianinha county, Brazil

Além de influenciar o crescimento corpóreo, o hormônio do crescimento, ou somatotrófico, desempenha importante papel no metabolismo, composição corporal, perfil lipídico, estado cardiovascular e longevidade. Seu controle é multi-regulado por hormônios, metabólitos e peptídeos hipotalâmicos. Dados sobre a Deficiência Isolada de GH (DIGH) obtidos a partir da descrição da mutação IVS1+1G®A no gene do receptor do hormônio liberador do GH (GHRH-R) em indivíduos da cidade de Itabaianinha, SE, são revisados. São abordadas novas perspectivas sobre o modelo de resistência ao GHRH, a importância do GHRH no controle da secreção de GH, a freqüência das mutações do gene do GHRH-R, a relevância diagnóstica do IGF-I e os achados metabólicos, cardiovasculares e de qualidade de vida nestes indivíduos.

[Growth or somatotrophic hormone: new perspectives in isolated GH deficiency after description of the mutation in the GHRH receptor gene in individuals of Itabaianinha County, Brazil]. / Hormônio do crescimento ou somatotrófico: novas perspectivas na deficiência isolada de GH a partir da descrição da mutação no gene do receptor do GHRH nos indivíduos da cidade de Itabaianinha, Brasil.

In addition to stimulating body growth, growth or somatotrophic hormone plays an important role in metabolism, body composition, lipid profile, cardiovascular status and longevity. Its control is multiregulated by hormones, metabolites and hypothalamic peptides. Obtained data of the isolated growth hormone deficiency (IGHD) after the description of the IVS1+1G-->A GHRH receptor gene mutation in individuals of Itabaianinha County are reviewed. New perspectives about the growth hormone resistance model, the importance of GHRH in the control of GH secretion, the frequency of GHRH-R gene mutations, the diagnostic relevance of IGF-I and the metabolic, cardiovascular and quality of life findings are approached.

Batch xylitol production from wheat straw hemicellulosic hydrolysate using Candida guilliermondii in a stirred tank reactor.

Batch production of xylitol from the hydrolysate of wheat straw hemicellulose using Candida guilliermondii was carried out in a stirred tank reactor (agitation speed of 300 rpm, aeration rate of 0.6 vvm and initial cell concentration of 0.5 g l(-1)). After 54 h, xylitol production from 30.5 g xylose l(-1) reached 27.5 g l(-1), resulting in a xylose-to-xylitol bioconversion yield of 0.9 g g(-1) and a productivity of 0.5 g l(-1) h(-1).

Metabolic behavior of immobilized Candida guilliermondii cells during batch xylitol production from sugarcane bagasse acid hydrolyzate.

Candida guilliermondii cells, immobilized in Ca-alginate beads, were used for batch xylitol production from concentrated sugarcane bagasse hydrolyzate. Maximum xylitol concentration (20.6 g/L), volumetric productivity (0.43 g/L. h), and yield (0.47 g/g) obtained after 48 h of fermentation were higher than similar immobilized-cell systems but lower than free-cell cultivation systems. Substrates, products, and biomass concentrations were used in material balances to study the ways in which the different carbon sources were utilized by the yeast cells under microaerobic conditions. The fraction of xylose consumed to produce xylitol reached a maximum value (0.70) after glucose and oxygen depletion while alternative metabolic routes were favored by sub-optimal conditions.