Recovery of Hemicelluloses from Wood Hydrolysates by Membrane Filtration

Hemicelluloses are among the most important natural resources that contain polysaccharides. In this study the separation and purification of hemicelluloses from water extraction liquors containing wood hemicelluloses, lignin compounds and monosaccharide by using membrane filtration was investigated. The isolation of the hemicelluloses from the wood hydrolysates was performed in two steps: concentration of high molar mass hemicelluloses by ultrafiltration and separation of low molar mass hemicelluloses from monomeric sugars using tight ultrafiltration membranes. The purification of the retained hemicelluloses was performed by diafiltration. During the filtration experiments, the permeate flux through ultrafiltration and tight ultrafiltration membranes was relatively high. The fouling ability of the used membranes was relatively low. In our experiments, the retention of hemicelluloses using two filtration steps was almost complete. The separation of monosaccharides from hemicelluloses was relatively high and the purification of hemicelluloses by diafiltration was highly efficient. The separation of lignin from hemicelluloses was partially achieved. Diafiltration showed potential to purify retained hemicelluloses from lignin and other organics. The best separation of lignin from hemicelluloses in the first filtration step was obtained using the UC005 membrane. The GE-5 and ETNA01PP membranes showed potential to purify and separate lignin from hemicelluloses. However, the feed solution of the second filtration stages (from different ultrafiltration membranes) affected the permeate flux and the separation of various extracted compounds from hemicelluloses. The GE-5 and ETNA01PP membranes gave the efficient purification of the hemicelluloses when using diafiltration. Separation of degraded xylan from glucomannan (primary spruce hemicelluloses) was also possible using membrane filtration. The best separation was achieved using the GE-5 membrane. The retention of glucomannan was three times higher than xylan retention.

Effects of the partial removal of wood hemicelluloses on the properties of kraft pulp

The objective of this work was to study the effects of partial removal of wood hemicelluloses on the properties of kraft pulp.The work was conducted by extracting hemicelluloses (1) by a softwood chip pretreatment process prior to kraft pulping, (2) by alkaline extraction from bleached birch kraft pulp, and (3) by enzymatic treatment, xylanase treatment in particular, of bleached birch kraft pulp. The qualitative and quantitative changes in fibers and paper properties were evaluated. In addition, the applicability of the extraction concepts and hemicellulose-extracted birch kraft pulp as a raw material in papermaking was evaluated in a pilot-scale papermaking environment. The results showed that each examined hemicellulose extraction method has its characteristic effects on fiber properties, seen as differences in both the physical and chemical nature of the fibers. A prehydrolysis process prior to the kraft pulping process offered reductions in cooking time, bleaching chemical consumption and produced fibers with low hemicellulose content that are more susceptible to mechanically induced damages and dislocations. Softwood chip pretreatment for hemicellulose recovery prior to cooking, whether acidic or alkaline, had an impact on the physical properties of the non-refined and refined pulp. In addition, all the pretreated pulps exhibited slower beating response than the unhydrolyzed reference pulp. Both alkaline extraction and enzymatic (xylanase) treatment of bleached birch kraft pulp fibers indicated very selective hemicellulose removal, particularly xylan removal. Furthermore, these two hemicellulose-extracted birch kraft pulps were utilized in a pilot-scale papermaking environment in order to evaluate the upscalability of the extraction concepts. Investigations made using pilot paper machine trials revealed that some amount of alkalineextracted birch kraft pulp, with a 24.9% reduction in the total amount of xylan, could be used in the papermaking stock as a mixture with non-extracted pulp when producing 75 g/m2 paper. For xylanase-treated fibers there were no reductions in the mechanical properties of the 180 g/m2 paper produced compared to paper made from the control pulp, although there was a 14.2% reduction in the total amount of xylan in the xylanase-treated pulp compared to the control birch kraft pulp. This work emphasized the importance of the hemicellulose extraction method in providing new solutions to create functional fibers and in providing a valuable hemicellulose co-product stream. The hemicellulose removal concept therefore plays an important role in the integrated forest biorefinery scenario, where the target is to the co-production of hemicellulose-extracted pulp and hemicellulose-based chemicals or fuels.

Utilizing hemicelluloses – the first step : controlled extraction with hot water

Människor utnyttjar ofta kemi mångsidigt i sitt vardagliga liv utan att närmare tänka på detaljerna. Nuförtiden kan man framställa en ökande mängd av produkter ur förnybara råmaterial och en av de mest mångsidiga nybara råmaterialet i Norden är barrträd. Den lyriska lägerelden eller spiselden och möbler av ved samt papper är en väsentlig del av vardagen. Också livsmedel och läkemedel kan innehålla föreningar ur ved. Ved som råmaterial består av tre huvudkomponenten: cellulosa, som är uppbyggd av druvsockermolekyler är en långkedjad, oförgrenad polymer; lignin, som sammanhåller fibrerna i vedmaterialet som lim samt hemicellulosor, som ofta är uppbyggda av olika sockerarter och är en förgrenad polymer. Följaktligen består vedmaterialet av 70 % socker. I detta arbete har vi koncentrerat på i hemicellulosa och dess extraktion ur gran, samt bestämning av hemicellulosans egenskaper. Den slutliga målsättningen i forskningen var att skapa nya produkter ur gran. Forskning i extraktionens hemligheter eller hur hemicellulosa kan effektivt extraheras i den önskade formen kräver nya typers experimentellasanläggningar och experiment samt matematisk modellering. Den långkedjade hemicellulosan är lämplig för att användas t.ex. i skyddshinnor eller i livsmedel. Medel- och småmolekylär hemicellulosa kan användas som utgångsämne för framställning av bränslen, smörjmedel, sockersyror och alkoholer, av vilka xylitol är mest känd för alla pga hälsobefrämjande effekter. Det är utomordentligt viktigt ur miljöns och energiekonomins synvinkel att sträva efter effektivering av utnyttjandet av den värdefullaste och största naturtillgången, skogen i vårt land, med alla möjliga sätt. Resultaten av denna forskning utnyttjar avsevärt den växande, nya, på skogen baserande biobaseradeindustrin, som framställer nya spetsprodukter samt skapar nya arbetsplatser. ----------------------------------------------------- Ihmiset hyödyntävät usein huomaamattaan kemiaa monipuolisesti jokapäiväisessä elämässä. Nykyään kasvava määrä tuotteista kyetään valmistamaan uusiutuvista raaka-aineista ja yksi monipuolisimmista uusiutuvista luonnonvaroistamme pohjolassa ovat havupuut. Tunnelmallinen nuotio tai takkatuli ja puiset huonekalut sekä paperi ovat olennainen osa arkea. Myös elintarvikkeet ja lääkkeet voivat sisältää puusta peräisin olevia yhdisteitä. Puu materiaalina koostuu rakenteeltaan pääosin kolmesta osasta; selluloosasta, joka on rypälesokerista koostuva pitkäketjuinen haaroittumaton polymeeri, ligniinistä, joka toimii puun koossa pitävänä liima-aineena ja hemiselluloosasta, joka on useista eri sokereista rakentunut haaroittunut polymeeri. Näin ollen puusta 70 % on sokeria. Tässä työssä olemme keskittyneet hemiselluloosaan ja sen uuttamiseen kuusesta, sekä ominaisuuksien kartoittamiseen. Tutkimusaiheen lopullinen tavoite on luoda uusia tuotteita kuusesta. Uuton salojen tutkiminen eli miten hemiselluloosa saadaan tehokkaasti uutettua halutunlaisena vaatii uudenlaisia koelaitteistoja ja kokeita, sekä matemaattista mallintamista. Suurikokoinen hemiselluloosa on sopivaa käytettäväksi esimerkiksi suojakalvoissa tai elintarvikkeissa. Keskikokoista ja pienimolekyylistä hemiselluloosaa voidaan käyttää lähtöaineena valmistettaessa polttoaineita, voiteluaineita, sokerihappoja ja sokerialkoholeja, joista xylitoli on terveysvaikutustensa vuoksi kaikille tuttu. Niin ympäristömme kuin myös energiataloutemme kannalta on ensiarvoisen tärkeää pyrkiä kaikin keinoin tehostamaan maallemme arvokkaan, sekä luonnonvaroistamme yhden suurimman, metsän, vastuullista hyödyntämistä. Tämän tutkimuksen tulokset hyödyntävät merkittävästi maahamme nousevaa uutta metsään pohjautuvaa biojalostusteollisuutta, joka valmistaa uusia huipputuotteita sekä luo työpaikkoja.

Enhancement of ultrafiltration process by pretreatment in recovery of hemicelluloses from wood extracts

Hemicelluloses are potential raw material for several items produced in future wood-based biorefineries. One possible method for recovering hemicelluloses from wood extracts is ultrafiltration (UF). However, low filtration capacities and severe fouling restrict the use of tight UF membranes in the treatment of wood extracts. The lack of suitable commercial membranes creates a need for pretreatment which would decrease fouling and increase the filtration capacity. This thesis focuses on the evaluation of the possibility to improve the filtration capacity and decrease fouling with the pretreatment of wood extracts. Methods which remove harmful compounds and methods which degrade them are studied, as well as combinations of the methods. The tested pretreatments have an influence on both the concentration of different compounds and the molecular mass distribution of the compounds in the extract. This study revealed that in addition to which kind of compounds were removed, also the change in molecular size distribution affected the filtration capacity significantly. It was shown that the most harmful compounds for the filtration capacity of the hydrophobic 5 kDa membrane were the ones capable of permeating the membrane and fouling also the inner membrane structure. Naturally, the size of the most harmful compounds depends on the used UF membrane and is thus case-specific. However, in the choice of the pretreatment method, the focus should be on the removal of harmful compound sizes rather than merely on the total amount of removed foulants. The results proved that filtration capacity can be increased with both adsorptive and oxidative pretreatments even by hundreds of per cents. For instance, the use of XAD7 and XAD16 adsorbents increased the average flux in the UF of a birch extract from nearly zero to 107 kg/(m2h) and 175 kg/(m2h), respectively. In the treatment of a spruce extract, oxidation by pulsed corona discharge (PCD) increased the flux in UF from 46 kg/(m2h) to 158 kg/(m2h). Moreover, when a birch extract batch was treated with laccase enzyme, the flux in UF increased from 15 kg/(m2h) to 36 kg/(m2h). However, fouling was decreased only by adsorptive pretreatment while oxidative methods had a negligible or even negative impact on it. This demonstrates that filtration capacity and fouling are affected by different compounds and mechanisms. The results of this thesis show that filtration capacity can be improved and fouling decreased through appropriate pretreatment. However, the choice of the best possible pretreatment is case-specific and depends on the wood extract and the membrane used. Finding the best option requires information on the extract content and membrane characteristics as well as on the filtration performance of the membrane in the prevailing conditions and a multivariate approach. On the basis of this study, it can be roughly concluded that adsorptive pretreatment improves the filtration capacity and decreases fouling rather reliably, but it may lead to significant hemicellulose losses. Oxidation reduces the loss of valuable hemicelluloses and could improve the filtration capacity, but fouling challenges may remain. Combining oxidation with adsorptive pretreatment was not a solution for avoiding hemicellulose losses in the tested cases.

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.

Fermentation kinetics for xylitol production by a Pichia stipitis D-Xylulokinase mutant previously grown in spent sulfite liquor

Spent sulfite pulping liquor (SSL) contains lignin, which is present as lignosulfonate, and hemicelluloses that are present as hydrolyzed carbohydrates. To reduce the biological oxygen demand of SSL associated with dissolved sugars, we studied the capacity of Pichia stipitis FPL-YS30 (xyl3 Delta) to convert these sugars into useful products. FPL-YS30 produces a negligible amount of ethanol while converting xylose into xylitol. This work describes the xylose fermentation kinetics of yeast strain P.stipitis FPL-YS30. Yeast was grown in rich medium supplemented with different carbon sources: glucose, xylose, or ammonia-base SSL. The SSL and glucose-acclimatized cells showed similar maximum specific growth rates (0.146 h(-1)). The highest xylose consumption at the beginning of the fermentation process occurred using cells precultivated in xylose, which showed relatively high specific activity of glucose-6-phosphate dehydrogenase (EC 1.1.1.49). However, the maximum specific rates of xylose consumption (0.19 g(xylose)/g(cel) h) and xylitol production (0.059 g(xylitol)/g(cel) h) were obtained with cells acclimatized in glucose, in which the ratio between xylose reductase (EC 1.1.1.21) and xylitol dehydrogenase (EC 1.1.1.9) was kept at higher level (0.82). In this case, xylitol production (31.6 g/l) was 19 and 8% higher than in SSL and xylose-acclimatized cells, respectively. Maximum glycerol (6.26 g/l) and arabitol (0.206 g/l) production were obtained using SSL and xylose-acclimatized cells, respectively. The medium composition used for the yeast precultivation directly reflected their xylose fermentation performance. The SSL could be used as a carbon source for cell production. However, the inoculum condition to obtain a high cell concentration in SSL needs to be optimized.

An alternative application to the Portuguese agro-industrial residue: Wheat straw

The effects of alkaline treatments of the wheat straw with sodium hydroxide were investigated. The optimal condition for extraction of hemicelluloses was found to be with 0.50 mol/l sodium hydroxide at 55C for 2 h. This resulted in the release of 17.3% of hemicellulose (% dry starting material), corresponding to the dissolution of 49.3% of the original hemicellulose. The yields were determined by gravimetric analysis and expressed as a proportion of the starting material. Chemical composition and physico-chemical properties of the samples of hemicelluloses were elucidated by a combination of sugar analyses, Fourier transform infrared (FTIR), and thermal analysis. The results showed that the treatments were very effective on the extraction of hemicelluloses from wheat straw and that the extraction intensity (expressed in terms of alkali concentration) had a great influence on the yield and chemical features of the hemicelluloses. The FTIR analysis revealed typical signal pattern for the hemicellulosic fraction in the 1,200-1,000 cm(-1) region. Bands between 1,166 and 1,000 cm(-1) are typical of xylans.

Utilization of pineapple stem juice to enhance enzyme-hydrolytic efficiency for sugarcane bagasse after an optimized pre-treatment with alkaline peroxide

The enzymatic hydrolysis of sugarcane bagasse was investigated by treating a peroxide-alkaline bagasse with a pineapple stem juice, xylanase and cellulase. Pre-treatment procedures of sugarcane bagasse with alkaline hydrogen peroxide were evaluated and compared. Analyses were performed using 2(4) factorial designs, with pre-treatment time, temperature, magnesium sulfate and hydrogen peroxide concentration as factors. The responses evaluated were the yield of cellobiose and glucose released from pretreated bagasse after enzymatic hydrolysis. The results show that the highest enzymatic conversion was obtained for bagasse using 2% hydrogen peroxide at 60 degrees C for 16 h in the presence of 0.5% magnesium sulfate. Bagasse (5%) was treated with pineapple stem extract, which contains mixtures of protease and esterase, in combination with xylanase and cellulase. It was observed that the amount of glucose and cellobiose released from bagasse increased with the mixture of enzymes. It is believed that the enzymes present in pineapple extracts are capable of hydrolyze specific linkages that would facilitate the action of digesting plant cell walls enzymes. This increases the amount of glucose and other hexoses that are released during the enzymatic treatment and also reduces the amount of cellulase necessary in a typical hydrolysis. (C) 2010 Elsevier Ltd. All rights reserved.

Caracterização da ontogenética do pau-brasil

As ações de reflorestamento com o pau-brasil (Caesalpinia echinata Lam.) depende de informações de suas características ecofisiológicas sujeitas às variações ontogenéticas e ambientais. O objetivo desse trabalho foi caracterizar alguns aspectos morfológicos, anatômicos, fisiológicos e estruturais de parede celular de C. echinata nas fases juvenil, jovem e adulto em condições naturais em um fragmento da Floresta Atlântica. Foi analisada a biometria, concentração de nutrientes e dos pigmentos cloroplastídicos dos foliólulos, anatomia foliar e do xilema secundário do caule e a constituição dos polímeros estruturais de parede celular. Os indivíduos juvenis localizados no estrato inferior da floresta se destacaram pela maior área foliar específica e maiores teores de pigmentos cloroplastídicos bem como pelas maiores dimensões de suas células guardas associado às maiores concentrações de K e Ca foliar. Estruturalmente, os indivíduos juvenis apresentaram menores elementos de vasos e teores de lignina. Os indivíduos jovens apresentaram valores intermediários das variáveis analisadas. Já os indivíduos adultos, cujas copas alcançavam o dossel, se destacaram pelo maior espessamento do limbo, da cutícula e do parênquima lacunoso, teor de água foliar, densidade estomática e teor de lignina foliar e caulinar cuja capacidade de síntese foi associada ao maior teor de P foliar. O conteúdo de celulose foliar e caulinar não variou entre as diferentes fases ontogenéticas. As hemiceluloses são do tipo xilanos com possibilidade de presença de xiloglucano dada a maior fração de xilose (±12% MS) e galactose (±1% MS). A glucose foi o monossacarídeo mais representativo (±40% MS) sem diferenças ontogenéticas. As diferenças morfológicas, anatômicas, fisiológicas e estruturais parecem, também, estarem sob controle da irradiância mais intensa na copa dos indivíduos adultos. Os resultados denotam que o plantio consorciado com espécies de crescimento rápido seja a melhor ação para o reflorestamento de C. echinata.

Study of thermochemical treatments of cork in the 150-400ºC range using colour analysis and Ftir spectroscopy

A study of chemical transformations of cork during heat treatments was made using colour variation and FTIR analysis. The cork enriched fractions from Quercus cerris bark were subjected to isothermal heating in the temperature range 150–400 ◦C and treatment time from 5 to 90 min. Mass loss ranged from 3% (90 min at 150 ◦C) to 71% (60 min at 350 ◦C). FTIR showed that hemicelluloses were thermally degraded first while suberin remained as the most heat resistant component. The change of CIE-Lab parameters was rapid for low intensity treatments where no significant mass loss occurred (at 150 ◦C L* decreased from the initial 51.5 to 37.3 after 20 min). The decrease in all colour parameters continued with temperature until they remained substantially constant with over 40% mass loss. Modelling of the thermally induced mass loss could be made using colour analysis. This is applicable to monitoring the production of heat expanded insulation agglomerates.

Olive pomace pretreatments to enhance its valorisation by solid-state fermentation

Dissertação de mestrado em Bioengenharia

Fermentação da gelose pelas bacterias anaerobias

1 - Anaerobic bacteria of the Clostridium genus acidify mineral media without when agar is added. 2 - Acidulation results from the attack on the agar as a source of carbon. 3 - The quantity of CO² produced by the decomposition of the agar is approximately that obtained with soil bacteria as shown by Waksmann and Diehm working with hemicelluloses. Although galactone is less atacked than the other hemicelluloses the acidity produced is sufficient to disturb the fermentation tests in semi-solid media with agar. 4 - The acidulation of Spray's sugar-free control medium is probably due to the decomposition of the agar by anaerobes. The acidity produced may interfere with the acidity of the fermentation of the sugar in Spray's test or may be added to it, thus giving a false indication of the real acidity.

Kalvoerotusprosessin esikäsittely

Työn tavoitteena oli tutkia kiertoveden puhdistamiseen käytetyn kalvosuodatuksen esikäsittelymenetelmien vaikutusta ultrasuodatukseen. Suodatettava vesi oli kierrätyskuitua käyttävän paperikoneen kiekkosuotimen kirkassuodosta. Koeajon loppuosassa käytettiin biokirkassuodosta. Paperitehtaassa oli suljettu vesikierto. Pilot-laitteistona käytettiin Metso PaperChemin OptiFilter CR550/40 ultrasuodatusyksikköä. Testatut kalvot olivat C30 kalvoja. Suodatuksissa seurattiin esikäsittelymenetelmien vaikutusta permeaatin määrään ja laatuun. Ultrasuodatus tehtiin sekä ilman esikäsittelyä että lamelliselkeytin esikäsittelynä. Esikäsittelykemikaalina käytettiin talkkia. Ultrasuodatuksissa havaittiin, että suodatuslämpötilalla oli suora vaikutus kapasiteettiin. Lämpötila vaikutti lineaarisesti kirkassuodoksen ja biokirkassuodoksen permeaattivuohon. Lämpötila nostettiin 55oC:een suodatuslämpötilasta 35 – 40oC. Korkeassa lämpötilassa kirkassuodos oli kapasiteetiltaan suurempi kuin biokirkassuodos. Konsentrointiajoista nähtiin, että ultrasuodatuksen syötön konsentraatioaste vaikutti kirkassuodoksen permeaattivuohon. Syötön konsentraatioastetta lisättäessä kapasiteetti laski. Konsentrointiajoja vertailtaessa havaitaan, että ilman esikäsittelyä vuo laski konsentroitaessa nopeammin kuin suodatuksissa, joissa käytettiin esikäsittelyä. Lamelliselkeyttimellä ja talkilla esikäsitelty suodatus laski kapasiteettia vähiten. Konsentraatioasteen nostaminen lisäsi konsentraatin kemiallista hapenkulutusta. Konsentraatin viskositeetti nousi konsentraatioastetta lisättäessä. Analyysitulosten perusteella saadaan ultrasuodatuskalvoille tyypilliset reduktiot. COD-reduktio oli 10 – 30 %, anionisuuden 40 – 70 % ja värin 80 – 90 %. Vuon kanssa korreloivat pH, uuteaine, ligniini, viskositeetti ja hemiselluloosat.

Acetylation of cell wall is required for structural integrity of the leaf surface and exerts a global impact on plant stress responses.

The epidermis on leaves protects plants from pathogen invasion and provides a waterproof barrier. It consists of a layer of cells that is surrounded by thick cell walls, which are partially impregnated by highly hydrophobic cuticular components. We show that the Arabidopsis T-DNA insertion mutants of REDUCED WALL ACETYLATION 2 (rwa2), previously identified as having reduced O-acetylation of both pectins and hemicelluloses, exhibit pleiotrophic phenotype on the leaf surface. The cuticle layer appeared diffused and was significantly thicker and underneath cell wall layer was interspersed with electron-dense deposits. A large number of trichomes were collapsed and surface permeability of the leaves was enhanced in rwa2 as compared to the wild type. A massive reprogramming of the transcriptome was observed in rwa2 as compared to the wild type, including a coordinated up-regulation of genes involved in responses to abiotic stress, particularly detoxification of reactive oxygen species and defense against microbial pathogens (e.g., lipid transfer proteins, peroxidases). In accordance, peroxidase activities were found to be elevated in rwa2 as compared to the wild type. These results indicate that cell wall acetylation is essential for maintaining the structural integrity of leaf epidermis, and that reduction of cell wall acetylation leads to global stress responses in Arabidopsis.