Phenols are priority pollutants that are commonly found in a large number of industrial wastewaters. Different processes are currently available for the elimination of phenol from wastewater but present some disadvantages like low efficiency, high energy-consumption, the necessity of acclimatisation of the sludges or the limitation of the treatment capacity. The need to find alternatives has made the enzymatic processes a good option. In the last two decades, several processes were implemented with different enzymes from plants and microorganisms, including peroxidases from several sources, as the horseradish peroxidase.Also, different enzyme configurations, free or immobilised enzyme and different supports for immobilisation have been studied. Substantial attention has been devoted to the covalent immobilisation of enzymes on porous insoluble supports such as glass, alumina, silica, and chitosan.The main novelty of this work is the utilisation of a torus reactor for the removal of organic contaminants from wastewaters. This reactor, which can be considered as a loop reactor, presents some advantages over other stirred tank reactors.The goal of this work is the study of the hydrodynamics of a torus reactor for its further application in the enzymatic elimination of phenol and the coupling of the kinetics and the modelisation.In a first step, the enzymatic elimination of phenol was experimentally studied in the torus reactor. In order to compare the performances, several assays were also carried out with a stirred reactor. A high degree of conversion was obtained for the enzymatic elimination of phenol in both reactors with the tested quantities of phenol. It was concluded that, keeping a ratio of 1:1 between the phenol and the H2O2 initial molar quantities, the highest final reaction conversion was obtained. Using the torus reactor was obtained 97% of phenol conversion when the optimal concentrations of substrates were usedIn order to improve economically the process, the enzyme should be used in a continuous regime over a long time period to exploit it completely. For this reason it was necessary to immobilise the enzyme. This work presents a new configuration that has never been tested: the horseradish peroxidase supported on Eupergit.In a second step, the characterisation using the CFD of the flow-field in a torus reactor of 100 ml, similar to the experimental reactor, was carried out for two different configurations, batch and continuous operating modes. Moreover, the scale-up of the volume of the torus reactor was carried out using CFD for a 300 ml reactor.Finally, the enzymatic reaction of phenol with the HRP was modelled using the CFD coupled to the kinetic model of the enzymatic reaction to the flow simulation. These results allowed the possibility of optimising and scaling-up the process using the CFD modelisation. / Los compuestos fenólicos son contaminantes prioritarios que se encuentran comúnmente en una gran cantidad de efluentes industriales. Diferentes procesos están disponibles actualmente para la eliminación de fenol desde dicho efluentes pero los mismos presentan algunas desventajas como pueden ser una baja eficiencia, un mayor consumo de energía, la producción de lodos conteniendo hierro o limitaciones en la capacidad de tratamiento. La necesidad de encontrar alternativas a estos problemas ha hecho del proceso enzimático una buena opción. En las últimas dos décadas, varios procesos han sido implementados utilizando diferentes enzimas extraídas de plantas y microorganismos como pueden ser las peroxidasas de diversas fuentes, incluyendo la horseradish peroxidasa.Diferentes configuraciones de enzimas, libre e inmovilizada y diferentes soportes para la inmovilización han sido también estudiados. Sustancial atención ha sido dedicada a la inmovilización de enzimas por enlace covalente sobre soportes porosos insolubles tales como vidrio, aluminio, sílice y chitosan. El objetivo de este trabajo es el estudio de la hidrodinámica dentro de un reactor tórico para su posterior aplicación en la eliminación enzimática de fenol y el acople entre las cinéticas y la modelización.En una primera etapa, la eliminación enzimática de fenol es estudiada experimentalmente en el reactor tórico. Con el objetivo de comparar el rendimiento de dicho reactor, varios ensayos se realizaron en un reactor agitado tradicional. Un alto grado de conversión de fenol ha sido obtenido para la eliminación enzimática de fenol en ambos reactores para las cantidades estudiadas de fenol. Ha sido observado que es necesario mantener una relación de 1:1 entre la concentración inicial de fenol y la de peróxido de hidrógeno para lograr la mayor conversión de fenol. Usando el reactor tórico ha sido obtenido un 97% de conversión de fenol cuando las concentraciones óptimas de substratos y enzimas fueron utilizados. Con el objetivo de mejorar económicamente el proceso y hacerlo factible para su uso a escala industrial, la enzima debería ser utilizada en un proceso en continuo sobre un largo período de tiempo para explotarla completamente. Por esta razón, ha sido necesario inmovilizar la enzima. Este trabajo muestra una nueva configuración que no ha sido aún probada: la horseradish peroxidase soportada en Eupergit. Asimismo, la caracterización usando CFD del campo de flujo de un reactor tórico similar al experimental de 100 ml ha sido realizada para un reactor trabajando de forma batch y continua. Un escalado en el volumen del reactor tórico ha sido realizado utilizando CFD para un reactor de 300 ml. Finalmente, la reacción enzimática de fenol con HRP has sido modelada acoplando el modelo cinético obtenido experimentalmente con las simulaciones del campo de flujo dentro del reactor. Estos resultados permitirán la optimización y el escalado del proceso usando CFD.

Chicken feathers are the most abundant keratinous biomass in the world. Disposal of thehuge and increasing volume of waste feathers presents as a major concern for poultryindustry. On the other hand, energy and material recovery of this valuable protein sourceis an important issue for organic solid waste treatment and bioenergy generation.Anaerobic digestion is an environmentally and economically promising alternativeprocess for biogas production of waste feathers.In this study in order to enhance the methane potential of batch anaerobic digestion ofchicken feathers this waste was treated by various kinds of pretreatments includingthermal, thermo-chemical, enzymatic, thermo-enzymatic and chemo-enzymatic methods.Also the effect of different treatment conditions on the methane yield was investigated.As a whole, thermo-chemical pretreatment with lime (Ca(OH)2) rendered the mostsignificant effect on enhancement of the chicken feathers methane potential. In particularlime treated triplicate samples under treatment condition of 40g TS feather/l water, 0.1gCa (OH)2 /g TS feather, 100°C and 30 min produced the highest amount of methane (anaverage maximum volume of 480 Nml/g VS, which is about 96.8% of the theoreticalmethane potential of protein), during 50 days of anaerobic incubation. Increasing theoperational parameters such as feather concentration, lime loading, temperature andreaction time improved the feathers solublisation resulting in a higher soluble chemicaloxygen demand (SCOD) concentration of the samples but inserted negative impacts onthe anaerobic digestion performance. Although other pretreatment methods improved theSCOD concentrations of the feathers too, compared to the lime treatment those methodsdidn’t show considerable effects on the enhancement of methane yield from the chickenfeathers. Thermo-enzymatic, enzymatic, and thermal pretreated triplicate samplesproduced an average maximum of 185 Nml/g VS, 154 Nml/g VS, and 143 Nml/g VS(37.3%, 31%, 28.8% of the theoretical methane potential) respectively, during 33 days of50 days of anaerobic incubation. Especially, chemo-enzymatic pretreated sample showednegative methane potential of only 41 Nml/g VS, i.e. 8% of the theoretical methanepotential. Consequently, lime pretreatment under the above recommended conditions canbe suggested for hydrolysis of chicken feathers to achieve significant enhancement of itsmethane potential.

<p>Cellulosans tillgänglighet och reaktivitet är nyckelparametrar vid framställning av regenererad cellulosa och cellulosaderivat. Det är välkänt att på grund av cellulosans kristallina struktur är tillgängligheten begränsad för lösningsmedel och olika reagens. Till exempel kan en inhomogen substitution av hydroxylgrupperna i cellulosakedjan resultera i cellulosaderivat av sämre kvalitet. Baserat på detta har en del av arbetet i denna studie fokuserat på att förbättra cellulosans tillgänglighet och reaktivitet genom att studera effekten av olika enzymatiska behandlingar med monokomponent endoglukanaser. Resultaten visar att närvaron av en cellulosabindande domän fyller en viktig funktion för att öka cellulosans reaktivitet, men strukturen för den katalytiska domänen visade sig ha den största inverkan på cellulosans tillgänglighet. I kompletterande studier har även effekten av en mekanisk förbehandling i kombination med enzymatisk behandling utvärderats. Kombinationen av förbehandlingarna resulterade i en positiv effekt, cellulosans reaktivitet kunde ökas i större omfattning.</p><p>I dag används huvudsakligen dissolvingmassor som råvara vid framställning av cellulosaregenerat och cellulosaderivat. Kraven för dessa s.k. specialmassor är högt cellulosainnehåll samt lågt hemicellulosa- respektive lignininnehåll. På grund av dessa specifika krav är produktionskostnaderna för dessa massor högre än konventionella sulfatmassor. Den andra delen av studien har därför fokuserat på möjligheten att använda dessa sulfatmassor som dissolvingmassa. Det har visats att kombinationen av enzymatiska behandlingar med monokomponent endoglukanas och xylanas följt av ett alkaliskt steg kan resultera i massor där kraven uppfylls med avseende på cellulosans reaktivitet, och cellulosa- respektive hemicellulosainnehåll.</p> / <p>The accessibility and reactivity of cellulose are key parameters on the manufacturing of cellulose derivatives and regenerated cellulose. It is well known that, due to the crystalline structure of cellulose, the accessibility of solvents and reagents is limited. For instance, an inhomogeneous substitution of the hydroxyl groups of the cellulose chain might lead to the production of derivatives of low quality. As a consequence, part of this work has focused on improving the accessibility and reactivity on cellulose by studying the effect of different monocomponent endoglucanases. It has been demonstrated that the presence of the cellulose-binding domain plays an important role on the enhancement of cellulose reactivity; however, the structure of the catalytic domain has been showed to have the highest influence on this parameter. Furthermore, the influence of mechanical treatment prior to enzymatic treatment has been examined. The combination of pretreatments showed a positive effect enhancing to a larger extent the cellulose reactivity.</p><p>Currently, dissolving-grade pulps are commonly used for the production of cellulose derivatives and regenerated cellulose. The requirements for these so-called “special pulps” are a high cellulose content and a low hemicelluloses and lignin content. As a result of these specific demands, the production costs of these pulps are higher than those of common kraft pulps. The second part of this work, therefore, has been focused on the study on the viability of converting kraft pulps into dissolving pulps. It has been demonstrated that the combination of enzymatic treatments using a monocomponent endoglucanase and a xylanase together with the addition of an alkaline step could fulfil the requirements of a commercial dissolving pulp in terms of cellulose reactivity and cellulose and hemicellulose content.</p>

Cette thèse présente des travaux concernant la modélisation mathématique de deux méthodes physiques existantes pour surmonter les barrières biologiques s’opposant à l’administration efficace de médicaments. Dans la première partie, plusieurs manières de modéliser l’électroporation sont exposées, aux échelles tissulaire et cellulaire. Des modèles phénoménologiques existants d’électroporation tissulaire sont présentés et comparés numériquement. Puis un modèle macroscopique d’électroporation est déduit d’un modèle d’électroporation cellulaire bien établi en utilisant des techniques d’homogénéisation. Dans la seconde partie, un nouveau modèle poroélastique est introduit pour décrire les écoulements dans un tissu biologique. Celui-ci prend en compte la dégradation tissulaire consécutive à un traitement par enzyme. Pour finir, un algorithme d’optimisation est proposé dans le but de déterminer un protocole optimal pour effectuer un traitement enzymatique. / This PhD thesis is devoted to the mathematical modeling and simulation of two existing physical methods to overcome the biological barriers to drug delivery. In the first part, several ways to model electroporation are considered, from the cell scale to the tissue scale. Existing phenomenological models of tissue electroporation are presented and numerically compared. Then a macroscopic model of electroporation is derived from a well-established model of cell elecroporation using homogenization techniques. In the second part, a new poroelastic model for the flows in biological tissues is presented to account for tissue degradation after an enzymatic treatment. To finish, an optimization algorithm is suggested in attempt to determine an optimal protocol when considering enzyme based therapies.

Cellulosans tillgänglighet och reaktivitet är nyckelparametrar vid framställning av regenererad cellulosa och cellulosaderivat. Det är välkänt att på grund av cellulosans kristallina struktur är tillgängligheten begränsad för lösningsmedel och olika reagens. Till exempel kan en inhomogen substitution av hydroxylgrupperna i cellulosakedjan resultera i cellulosaderivat av sämre kvalitet. Baserat på detta har en del av arbetet i denna studie fokuserat på att förbättra cellulosans tillgänglighet och reaktivitet genom att studera effekten av olika enzymatiska behandlingar med monokomponent endoglukanaser. Resultaten visar att närvaron av en cellulosabindande domän fyller en viktig funktion för att öka cellulosans reaktivitet, men strukturen för den katalytiska domänen visade sig ha den största inverkan på cellulosans tillgänglighet. I kompletterande studier har även effekten av en mekanisk förbehandling i kombination med enzymatisk behandling utvärderats. Kombinationen av förbehandlingarna resulterade i en positiv effekt, cellulosans reaktivitet kunde ökas i större omfattning. I dag används huvudsakligen dissolvingmassor som råvara vid framställning av cellulosaregenerat och cellulosaderivat. Kraven för dessa s.k. specialmassor är högt cellulosainnehåll samt lågt hemicellulosa- respektive lignininnehåll. På grund av dessa specifika krav är produktionskostnaderna för dessa massor högre än konventionella sulfatmassor. Den andra delen av studien har därför fokuserat på möjligheten att använda dessa sulfatmassor som dissolvingmassa. Det har visats att kombinationen av enzymatiska behandlingar med monokomponent endoglukanas och xylanas följt av ett alkaliskt steg kan resultera i massor där kraven uppfylls med avseende på cellulosans reaktivitet, och cellulosa- respektive hemicellulosainnehåll. / The accessibility and reactivity of cellulose are key parameters on the manufacturing of cellulose derivatives and regenerated cellulose. It is well known that, due to the crystalline structure of cellulose, the accessibility of solvents and reagents is limited. For instance, an inhomogeneous substitution of the hydroxyl groups of the cellulose chain might lead to the production of derivatives of low quality. As a consequence, part of this work has focused on improving the accessibility and reactivity on cellulose by studying the effect of different monocomponent endoglucanases. It has been demonstrated that the presence of the cellulose-binding domain plays an important role on the enhancement of cellulose reactivity; however, the structure of the catalytic domain has been showed to have the highest influence on this parameter. Furthermore, the influence of mechanical treatment prior to enzymatic treatment has been examined. The combination of pretreatments showed a positive effect enhancing to a larger extent the cellulose reactivity. Currently, dissolving-grade pulps are commonly used for the production of cellulose derivatives and regenerated cellulose. The requirements for these so-called “special pulps” are a high cellulose content and a low hemicelluloses and lignin content. As a result of these specific demands, the production costs of these pulps are higher than those of common kraft pulps. The second part of this work, therefore, has been focused on the study on the viability of converting kraft pulps into dissolving pulps. It has been demonstrated that the combination of enzymatic treatments using a monocomponent endoglucanase and a xylanase together with the addition of an alkaline step could fulfil the requirements of a commercial dissolving pulp in terms of cellulose reactivity and cellulose and hemicellulose content. / QC 20101117

This thesis deals with the preparation of chemically and/or enzymatically modified cellulose. This modification can be either irreversible or reversible. Irreversible modification is used to prepare cellulose derivatives as end products, whereas reversible modification is used to enhance solubility in the preparation of regenerated cellulose. The irreversible modification studied here was the preparation of carboxymethyl cellulose (CMC) using extended mercerization of a spruce dissolving pulp. More specifically the parameters studied were the effect of mercerization at different proportions of cellulose I and II in the dissolving pulp, the concentration of alkali, the temperature and the reaction time. The parameters evaluated were the degree of substitution, the filterability and the amount of gel obtained when the resulting CMC was dissolved in water. Molecular structures of CMC and its gel fractions were analysed by using NIR FT Raman spectroscopy. It was found that the alkali concentration in the mercerization stage had an extensive influence on the subsequent etherification reaction. FT Raman spectra of CMC samples and their gel fractions prepared with low NaOH concentrations (9%) in the mercerization stage indicated an incomplete transformation of cellulose to Na-cellulose before carboxymethylation to CMC. Low average DS values of the CMC, i.e. between 0.42 and 0.50 were obtained. Such CMC dissolved in water resulted in very thick and semi solid gum-like gels, probably due to an uneven distribution of substituents along the cellulose backbone. FT Raman spectra of CMC samples and their gel fractions mercerized at higher alkaline concentration, i.e. 18.25 and 27.5% in the mercerization stage, indicated on the other hand a complete transformation of cellulose to Na-cellulose before carboxymethylation to CMC. Higher average DS values of the CMC, i.e. between 0.88 and 1.05 were therefore obtained. When dissolved in water such CMC caused gel formation especially when prepared from dissolving pulp with a high fraction of cellulose II. The reversible modification studied was the dissolution of cellulose in NaOH/ZnO. Here the effect of enzyme pretreatment was investigated by using two mono-component enzymes; namely xylanase and endoglucanase, used in consecutive stages. It was found that although the crystallinity and the specific surface area of the dissolving pulp sustained minimal change during the enzymatic treatment; the solubility of pulp increased in a NaOH/ZnO solution from 29% for untreated pulp up to 81% for enzymatic pretreated pulp. / Baksidetext Cellulose can be chemically and/or enzymatically modified. Irreversible modification is used to prepare cellulose derivatives as end products, reversible modification to enhance solubility in the preparation of regenerated cellulose. The irreversible modification studied here was the preparation of carboxymethyl cellulose (CMC) using extended mercerization of a spruce dissolving pulp. More specifically the parameters studied were the effect of mercerization at different proportions of cellulose I and II in the dissolving pulp, the concentration of alkali, the temperature and the reaction time. It was found that the alkali concentration in the mercerization stage had an extensive influence on the subsequent etherification reaction. The content of cellulose II had little effect on degree of substitution (DS) at low NaOH concentration, but tended to decrease DS at higher NaOH concentration in both cases compared with cellulose I. It was also found that the content of cellulose II correlates with the gel formation obtained when the CMC is dissolved in water. The reversible modification studied was the dissolution of cellulose in NaOH/ZnO. Here the effect of enzyme pretreatment was investigated by using two mono-component enzymes; namely xylanase and endoglucanase, used in consecutive stages. It was found that the solubility of pulp increased in a NaOH/ZnO solution from 29% for untreated pulp up to 81% for enzymatic pretreated pulp.

Commercial carrot processors produce up to 175,000 tons of carrot waste annually. Carrot Mash (CM) is the term referring to the waste by-product of peeled baby carrot processing. Transportation of carrot processing waste is expensive due to its high- water content (approx. 83-95%). High in bioactive compounds (carotenoids) and dietary fibers, it is expected that its conversion into a value-added by-product is of interest to the carrot processing industry. Hemicellulose-rich plant materials have proven to be a source of oligosaccharides, which are known for their beneficial prebiotic activity. The objectives of this research were to: 1) determine the effect of mechanical treatments on the extraction of water and bioactive compounds and evaluate the functional properties of carrot mash; 2) incorporate dried carrot mash into a beef patty and evaluate changes in pH, color, cooking yield, and texture; 3) apply an enzymatic treatment to carrot mash to promote the conversion of polysaccharides to oligosaccharides for prebiotic benefits. Mechanical separation of liquid and solid fractions by way of expeller pressing was efficient in extracting liquid while simultaneously increasing total solids by nearly 200%, the extraction of carotenoids by 1000%, and polyphenol content by nearly 97%. Mechanical treatments increased the fat binding capacity on average by 183% compared to untreated mash. The addition of unpressed carrot mash or expeller pressed carrot mash increased the cooking yield of a beef patty by 3-13% without significantly changing its textural properties. Enzymatically treating the carrot mash significantly increased the concentration of oligosaccharides up to 2.3%. These results suggest that carrot processing wastes can be physically and enzymatically modified and have an immense potential to be utilized as a functional ingredient in human food rather than being landfilled, composted or used as animal feed.