Enzymic saccharification of sugarcane bagasse pretreated by autohydrolysis-steam explosion

Pretreatment of bagasse by autohydrolysis at 200 degrees C for 4 min and explosive defibration resulted in the solubilization of 90% of the hemicellulose (a heteroxylan) and in the production of a pulp that was highly susceptible to hydrolysis by cellulases from Trichoderma reesei C-30 and QM 9414, and by a comercial preparation, Meicelase. Saccharification yields of 50% resulted after 24 h at 50 degrees C (pH 5.0) in enzymic digests containing 10% (w/v) bagasse pulps and 20 filter paper cellulase units (FPU). Saccharifications could be increased to more than 80% at 24 h by the addition of exogenous beta-glucosidase from Aspergillus niger. The crystallinity of cellulose in bagasse remained unchanged following autohydrolysis-explosion and did not appear to hinder the rate or extent of hydrolysis of cellulose. Autohydrolysis-exploded pulps extracted with alkali or ethanol to remove lignin resulted in lowere conversions of cellulose (28-36% after 25 h) than unextracted pulps. Alkali extracted pulps arising from autohydrolysis times of more than 10 min at 200 degrees C were less susceptible to enzymic hydrolysis than unextracted pulps and alkali-extracted pulps arising from short autohydrolysis times (e.g., 2 min at 200 degrees C). Autohydrolysis-explosion was as effective a pretreatment method as 0.25M NaOH (70 degrees C/2 h) both yielded pulps that resulted in high cellulose conversions with T. reesei cellulase preparations and Meicelase. Supplementation of T. reesei C-30 cellulose preparations with A. niger beta-glucosidases was effective in promoting the conversion of cellulose into glucose. A ration of FPU to beta-glucosidase of 1:1.25 was the minimum requirement to achieve more than 80% conversion of cellulose into glucose within 24 h. Other factors which influenced the extent of saccharification of autohydrolysis-exploded bagasse pulps were the enzyme-substrate ratio, the substrate concentration, and the saccharification mode.     

Effect of xylanases on peroxide bleachability of eucalypt (E. globulus) kraft pulp

Industrial eucalypt (E. globulus L.) kraft pulp was treated with two commercial xylanase preparations Ecopulp^® TX-200A and Pulpzyme^® HC (endo-1,4-β-xylanase activity; EC 3.2.1.8) and bleached by totally chlorine-free (TCF) three-stage hydrogen peroxide bleaching sequence, without oxygen pre-delignification. The effect of enzymatic stage on pulp properties and bleachability has been studied and compared with reference (control) pulps, processed without enzyme addition. The similar mode of enzymatic action was noted for both xylanase preparations. Final brightness of 86% ISO was achieved after complete bleaching. Direct bleaching effect caused pulp brightening (by 1.2–1.5% ISO) and delignification (by 7–10%) immediately after the enzymatic stage. The maximal bleach boosting was shown after the first peroxide stage and then diminished, despite the progressive increase in delignification over the control. The loss in efficiency of xylanase treatment by the end of peroxide bleaching was associated with specific behavior of xylan-derived chromophores, i.e., hexenuronic acids.     

Preparation and characterisation of methylcellulose from annual cardoon and juvenile eucalyptus

We have studied the preparation of methylcellulose from annual cardoon and juvenile eucalyptus. The high cellulose and low lignin contents of these plants make them potential alternative resources for cellulose derivation. Their high quality pulps were obtained by the Impregnation Rapid Steam Pulping (IRSP) process and Total Chloride Free (TCF) bleaching sequences using hydrogen peroxide. The bleached pulp was methylated twice in an isopropanol slurry at 60 °C for 22 h after mercerisation in 40% NaOH solution. Yields of water-soluble and alkali-soluble methylcellulose were determined by solvent extraction. Substitution patterns of methylcellulose were determined by ¹³C Nuclear Magnetic Resonance (NMR) spectroscopy. The intrinsic viscosities and solution properties of methylcellulose were measured in either 4% NaOH solution or dimethyl sulphoxide (DMSO). This study shows that annual cardoon and juvenile eucalyptus can produce high-quality methylcellulose, which can be used as alternative resources for the production of methylcellulose.     

TCF mill trial on softwood pulp with Korsäs thermostable and alkaline stable xylanase T6

Abstract: Use of hemicellulases, including xylanases, for delignification in the paper industry has been slowed down by the lack of large-scale availability of enzymes which are active at a high pH (above 8) and a high temperature (above 60°C), conditions prevailing in many bleaching processes. During the past years, acidic or neutral hemicellulases, working at temperatures below 60°C, were used in most mill experiments. The Korsäs T6 xylanase from Bacillus stearothermophilus , which is active at a pH above 9.0 and at a temperature above 65°C, was produced on a large scale in collaboration with Gist-brocades and was employed on a full scale mill trial to produce a Total Chlorine chemical-Free (TCF) pulp from softwood. The bleaching sequence used was (OO)BQQPP. where O stands for oxygen delignification. B for the enzymatic treatment, Q for the chelating agent step and P for the hydrogen peroxide step. The enzyme bleaching step was performed during a period of 4 h at 63 ± 1°C and pH 8.7 ± 0.1. The results of the mill trial show that the TCF pulp produced had a brightness of 78% ISO and, at the same time, it preserved the same strength properties as chlorine dioxide-bleached pulp. The saving of hydrogen peroxide was 20%. The results on brightness, strength and chemical saving of this first full scale trial with T6 xylanase indicate that, after optimization, a TCF bleaching sequence including an enzymatic step with a xylanase working at a high pH and a high temperature, such as T6 xylanase, can be used to produce a high-strength bleached pulp. The advantages of a high pH and a high temperature enzymatic bleaching step are discussed.     

Delignification of sugarcane bagasse using glycerol-water mixtures to produce pulps for saccharification

This paper describes the organosolv delignification of depithed bagasse using glycerol–water mixtures without a catalyst. The experiments were performed using two separate experimental designs. In the first experiment, two temperatures (150 and 190 °C), two time periods (60 and 240 min) and two glycerol contents (20% and 80%, v/v) were used. In the second experiment, which was a central composite design, the glycerol content was maintained at 80%, and a range of temperatures (141.7–198.3 °C) and time (23–277 min) was used. The best result, obtained with a glycerol content of 80%, a reaction time of 150 min and a temperature of 198.3 °C, produced pulps with 54.4% pulp yield, 7.75% residual lignin, 81.4% delignification and 13.7% polyose content. The results showed that high contents of glycerol tend to produce pulps with higher delignification and higher polyoses content in relation to the pulps obtained from low glycerol content reactions. In addition, the proposed method shows potential as a pretreatment for cellulose saccharification.     

Effect of hydrogen peroxide pretreatment on the structural features and the enzymatic hydrolysis of rice straw

Assessment was made to evaluate the effect of hydrogen peroxide pretreatment on the change of the structural features and the enzymatic hydrolysis of rice straw. Changes in the lignin content, weight loss, accessibility for Cadoxen, water holding capacity, and crystallinity of straw were measured during pretreatment to express the modification of the lignocellulosic structure of straw. The rates and the extents of enzymatic hydrolysis, cellulase adsorption, and cellobiose accumulation in the initial stage of hydrolysis were determined to study the pretreatment effect on hydrolysis. Pretreatment at 60 degrees C for 5 h in a solution with 1% (w/w) H(2)O(2) and NaOH resulted in 60% delignification, 40% weight loss, a fivefold increase in the accessibility for Cadoxen, an one times increase in the water-holding capacity, and only a slight decrease in crystallinity as compared with that of the untreated straw. Improvement on the pretreatment effect could be made by increasing the initial alkalinity and the pretreatment temperature of hydrogen peroxide solution. A saturated improvement on the structural features was found when the weight ratio of hydrogen peroxide to straw was above 0.25 g H(2)O(2)/g straw in an alkaline H(2)O(2) solution with 1% (w/w) NaOH at 32 degrees C. The initial rates and extents of hydrolysis, cellulase adsorption, and cellobiose accumulation in hydrolysis were enhanced in accordance with the improved structural features of straw pretreated. A four times increase in the extent of the enzymatic hydrolysis of straw for 24 h was attributed to the alkaline hydrogen peroxide pretreatment.     

A comparison of chemical pretreatment methods for improving saccharification of cotton stalks

The effectiveness of sulfuric acid (H(2)SO(4)), sodium hydroxide (NaOH), hydrogen peroxide (H(2)O(2)), and ozone pretreatments for conversion of cotton stalks to ethanol was investigated. Ground cotton stalks at a solid loading of 10% (w/v) were pretreated with H(2)SO(4), NaOH, and H(2)O(2) at concentrations of 0.5%, 1%, and 2% (w/v). Treatment temperatures of 90 degrees C and 121 degrees C at 15 psi were investigated for residence times of 30, 60, and 90 min. Ozone pretreatment was performed at 4 degrees C with constant sparging of stalks in water. Solids from H(2)SO(4), NaOH, and H(2)O(2) pretreatments (at 2%, 60 min, 121 degrees C/15 psi) showed significant lignin degradation and/or high sugar availability and hence were hydrolyzed by Celluclast 1.5L and Novozym 188 at 50 degrees C. Sulfuric acid pretreatment resulted in the highest xylan reduction (95.23% for 2% acid, 90 min, 121 degrees C/15 psi) but the lowest cellulose to glucose conversion during hydrolysis (23.85%). Sodium hydroxide pretreatment resulted in the highest level of delignification (65.63% for 2% NaOH, 90 min, 121 degrees C/15 psi) and cellulose conversion (60.8%). Hydrogen peroxide pretreatment resulted in significantly lower (p相似文献   

Laccase-HBT bleaching of eucalyptus kraft pulp: influence of the operating conditions

Different operating conditions (viz. pulp consistency, oxygen pressure and treatment time) in the biobleaching of eucalyptus kraft pulp with the laccase-HBT system was tested in order to describe their effect and normalize a biobleaching protocol. A high O(2) pressure (0.6MPa) was found to result in improved laccase-assisted delignification of the pulp. Also, a high pulp consistency (10%) and a short treatment time (2h) proved the best choices with a view to obtaining good pulp properties (kappa number and ISO brightness) under essentially mild conditions. The laccase-HBT treatment was found to result in slight delignification (in the form of a 20-27% decrease in kappa number); however, an alkaline extraction stage raised delignification to 41-45%, a much higher level than those obtained in the control tests (16-23%). Also, the use of hydrogen peroxide in the extraction stage resulted in improved brightness (14-19%), but in scarcely improved delignification (4-7%). Treating the pulp with the laccase-HBT system reduced the amount of hydrogen peroxide required for subsequent alkaline bleaching by a factor of 3-4 relative to control tests.     

Effect of pulping variables on the characteristics of oil-palm frond-fiber

Caustic pulping of oil-palm frond-fiber strands was conducted following a central composite design using a two-level factorial plan involving three pulping variables (temperature: 160-180 degrees C, time: 1-2 h, alkali charge: 20-30% NaOH). Responses of pulp properties to the process variables were analyzed using a statistical software (DESIGN-EXPERT). The results indicated that frond-fiber strands could be pulped with ease to about 35-45% yield. Statistically, the reaction time was not a significant factor while the influences of the treatment temperature and caustic charge were in general significantly relative to the properties of the resultant pulps.     

Evaluation of a combined brown rot decay–chemical delignification process as a pretreatment for bioethanol production from Pinus radiata wood chips

Wood chips of Pinus radiata softwood were biotreated with the brown rot fungus (BRF) Gloeophyllum trabeum for periods from 4 and 12 weeks. Biodegradation by BRF leads to an increase in cellulose depolymerization with increasing incubation time. As a result, the intrinsic viscosity of holocellulose decreased from 1,487 cm³/g in control samples to 783 and 600 cm³/g in 4- and 12-week decayed wood chips, respectively. Wood weight and glucan losses varied from 6 to 14% and 9 to 21%, respectively. Undecayed and 4-week decayed wood chips were delignified by alkaline (NaOH solution) or organosolv (ethanol/water) processes to produced cellulosic pulps. For both process, pulp yield was 5–10% lower for decayed samples than for control pulps. However, organosolv bio-pulps presented low residual lignin amount and high glucan retention. Chemical pulps and milled wood from undecayed and 4-week decayed wood chips were pre-saccharified with cellulases for 24 h at 50°C followed by simultaneous saccharification and fermentation (SSF) with the yeast Saccharomyces cerevisiae IR2-9a at 40°C for 96 h for bioethanol production. Considering glucan losses during wood decay and conversion yields from chemical pulping and SSF processes, no gains in ethanol production were obtained from the combination of BRF with alkaline delignification; however, the combination of BRF and organosolv processes resulted in a calculated production of 210 mL ethanol/kg wood or 72% of the maximum theoretically possible from that pretreatment, which was the best result obtained in the present study.     

Pulping cardoon (Cynara cardunculus) with peroxyformic acid (MILOX) in one single stage

In this work, depithed cardoon stalk (Cynara cardunculus) has been used with the objective of obtaining bleachable pulps. The material, once properly prepared, was subjected to one-step peroxyformic acid delignification. In order to study the process, a face-centred second order factorial design was developed which allowed the determination of the influences of four variables: concentrations of formic acid and hydrogen peroxide in the cooking liquor and the time and temperature of the treatment.

Empirical mathematical models have been obtained which predict the yield, kappa index, residual lignin content, and viscosity of the pulps. These models demonstrate that in general the delignification was extensive, producing pulps with kappa indexes less than 25 in the majority of cases, with good yields in the range of 45–60%. However, the pulps seem to have been degraded in the reaction media, as can be deduced from the low viscosity values found: 260–520 mL/g.     

NH(4)OH-KOH pulping mechanisms and kinetics of rice straw

The mechanisms and kinetics of NH(4)OH-KOH mixture pulping rice straw were studied. When aqueous ammonia was mixed with a small amount of caustic potash (ratio of 1:5), three distinct delignification phases were observed in the pulping process: a bulk delignification phase from the beginning of the cooking period to 100 degrees C, a supplementary delignification phase from 100 degrees C to 155 degrees C lasting a further 45 min, and a residual delignification phase until the end of the cooking period. There were two silica removal phases; the first phase was from the beginning of the cooking period to 100 degrees C and the second phase was from 100 degrees C to the end of the cooking period. The rate of delignification reaction was first order with respect to residual lignin and 0.3 order with respect to [OH(-)]. The silica removal was pseudo-first-order with respect to residual silica and 0.6 order with respect to [OH(-)]. The activation energies of the delignification and removal of silica reactions were 35.6 and 30.9 kJ/mol, respectively.     

Catalytic solvent delignification of agricultural residues: Organic catalysts

The effects of moderate-temperature aqueous butanol pretreatment of rice straw on several organic catalysts have been studied. Maximum delignification was obtained with an aromatic organic acid catalyst. Presoaking of straw in aqueous improved delignification, particularly in high solid-liquid rations. However, due to the difficulty of penetration of lignin into the solvent phase, an increase in the solid-liquid ratio appears to affect delignification. A two-stage catalytic solvent process involving presoaking at 80 degrees C followed by delignification at 120 degrees C resulted in 83% removal of lignin in aqueous-butanol, whereas 75% lignin removal was possible in ethanol. Higher reduction in the crystallinity was possible with etanol. Lignin produced by the solvent process offers desirable physical properties for polymer applications compared to Kraft lignin.     

Effect of oxygen delignification operating parameters on downstream enzymatic hydrolysis of softwood substrates

Enhanced oxygen delignification of softwood pulp samples (taken upstream and downstream of a commercial oxygen delignification unit) improved the initial rate of enzymatic saccharification and overall yield of monomeric sugars by 62-82% and 76-80%, respectively. Laboratory-scale experiments were used to examine the effect of a broad range of operating parameters (temperature, time, caustic concentration, and oxygen partial pressure) on the effectiveness of oxygen delignification. Using empirical models, kappa number (residual lignin content) was found to effectively predict final conversion to monomeric sugars. Application of oxygen delignification to sulfite mill knots resulted in smaller (20-25%) reduction in lignin content. However, using a combination of oxygen delignification and particle size reduction, up to 80% of the carbohydrate in the reject knots could be converted to fermentable sugars.     

Relationship between lignin structure and delignification degree in Pinus pinaster kraft pulps

This study examines the structure of residual and dissolved lignins from Pinus pinaster pulps obtained at different degrees of delignification by laboratory conventional kraft pulping. The cooking H factor was varied from 85 to 8049. The residual and dissolved lignin samples were characterised by elemental analysis, residual carbohydrate content, permanganate oxidation and 13C NMR spectroscopy. The reflectance factor of the pulps was also determined in order to tentatively correlate the delignification degree and residual lignin structure with the pulp colour. The obtained results confirmed that the delignification degree increases the condensation of the lignin structure, which might have an influence upon the observed increased pulp colour. The lack of selectivity of kraft pulping process in the case of more delignified pulps was also shown.     

Studies on Lignin

In comparison with the ethanolysis, the mercaptolysis of pine wood and pine ethanol lignin has been studied. The delignification was found to be almost complete when wood powder was cooked with the ethanolic hydrogen chloride containing 10% of ethyl mercaptan; while the same cooking without the mercaptan caused only 50% of delignification. Addition of 2% of mercaptan resulted more than 90% of delignification. As already reported briefly, from the mercaptolysis oil of pine ethanol lignin, 2-ethoxy-1-(4-hydroxy-3-methoxyphenyl)-propanone- (1), a thioether corresponding to Hibbert’s α-ethoxypropiovanillone, was isolated.     

Effect of structural features on enzyme digestibility of corn stover

Corn stover was pretreated with excess calcium hydroxide (0.5 g Ca(OH)2/g raw biomass) in non-oxidative and oxidative conditions at 25, 35, 45, and 55 degrees C. The enzymatic digestibility of lime-treated corn stover was affected by the change of structural features (acetylation, lignification, and crystallization) resulting from the treatment. Extensive delignification required oxidative treatment and additional consumption of lime (up to 0.17 g Ca(OH)2/g biomass). Deacetylation reached a plateau within 1 week and there were no significant differences between non-oxidative and oxidative conditions at 55 degrees C; both conditions removed approximately 90% of the acetyl groups in 1 week at all temperatures studied. Delignification highly depended on temperature and the presence of oxygen. Lignin and hemicellulose were selectively removed (or solubilized), but cellulose was not affected by lime pretreatment in mild temperatures (25-55 degrees C), even though corn stover was contacted with alkali for a long time, 16 weeks. The degree of crystallinity slightly increased from 43% to 60% with delignification because amorphous components (lignin, hemicellulose) were removed. However, the increased crystallinity did not negatively affect the 3-d sugar yield of enzymatic hydrolysis. Oxidative lime pretreatment lowered the acetyl and lignin contents to obtain high digestibility, regardless of crystallinity. The non-linear models for 3-d hydrolysis yields of glucan (Y(g)), xylan (Y(x)), and holocellulose (Y(gx)) were empirically established as a function of the residual lignin (L) for the corn stover pretreated with lime and air.     

Modification of wheat straw in a high-shear mixer

Wheat straw (Ws)was treated in a pilot-scale continuous mixer to disrupt the lignin-hemicellulose-cellulose (LHC) complex. An efficient and practical method was desired to remove lignin and hemicellulose (pentosans)rapidly and efficiently from the lignocellulose complex and to make the cellulose accessible to enzymatic hydrolysis. Milled WS in the presence of various chemicals in aqueous solutions was extruded from the mixer under several processing conditions. Chemicals used were sodium hydroxide (NaOH) sodium sulfide (Na(2)S), anthraquinone (AQ), anthrahydroquinone (AHQ), hexamethylenediamine (HMDA), hexamethylenetetramine (HMTA) hydrogen peroxide (H(2)O(2)), and ferrous ammonium sulfate (FAS), which were used alone and in selected combinations. Concomitantly, WS was treated in laboratory batches using similar reaction conditions, except for mixing and shearing. In extrusion treatments of WS at 20% concentration at 97 degrees C for 5.5 min with NaOH (15.7%, dry WS basis), NaOH (15.7%) + AHQ (0.3%), and NaOH (12.7%) + Na(2)S (5.0%), 64-72% of the WS lignin and 36-43% of the pentosans were removed from aqueously washed extrudates (residues). This compares with 46-56% and 23-27%, respectively, for batch treatments. AHQ and Na(2)S enhanced delignification. Cellulase treatment of the residues, which contained about 99%of the WS cellulose, converted 90-92%of the cellulose to glucose compared with 61-69%for the batch pretreatments. Treatments of WS with amines and H(2)O(2) (alone or combined with NaOH)were less effective for LHC disruption. In all instances the relatively high-shear extrusion treatments were superior to the laboratory-batch treatments.     

Alkaline subcritical water gasification of dairy industry waste (Whey)

The near-critical water gasification of dairy industry waste in the form of Whey, a product composed of mixtures of carbohydrates (mainly lactose) and amino acids such as glycine and glutamic acid, has been studied. The gasification process involved partial oxidation with hydrogen peroxide in the presence of NaOH. The reactions were studied over the temperature range from 300°C to 390°C, corresponding pressures of 9.5-24.5 MPa and reaction times from 0 min to 120 min. Hydrogen production was affected by the presence of NaOH, the concentration of H(2)O(2), temperature, reaction time and feed concentration. Up to 40% of the theoretical hydrogen gas production was achieved at 390°C. Over 80% of the Whey nitrogen content was found as ammonia, mainly in the liquid effluent.     

Allenic derivatives of nucleic acid components and their transformation products: a new class of biologically active nucleoside analogues

Reaction of adenine (1a) or cytosine (1b) with excess 1,4-dichloro-2-butyne catalyzed by K2CO3 in (CH3)2SO gave the 4-chloro-2-butynyl derivatives 2a and 2b. The latter were converted to the 4-hydroxy-2-butynyl compounds 3a and 3b by refluxing in 0.1 M HCl. Isomerization of 3a in 0.1 M NaOH at 100 degrees C for 1 h gave an equilibrium mixture of 3a and allene 4a. Pure 4a was obtained by column chromatography. Similarly, compound 3b was transformed/0.1 M NaOH, 20% aq. dioxane, 9 h, 100 degrees C/ to a mixture of 3b and 4b from which pure 4b was obtained by chromatography and crystallization. By contrast, reflux of 3a or 3b in 1 M NaOH in 50% aq. dioxane for 1 h afforded cyclized products - dihydrofuryl derivatives 5a and 5b. Hydrogenation of 4a and 5a gave 9-(4-hydroxybutyl)adenine (6a) and 9-(tetrahydro-2-furyl)adenine (7a), respectively. Scope and limitations of allenic isomerization in nucleic acid base series, spectroscopy and biological activity of the obtained products will be discussed.