Дисертації з теми "Glyoxol"

La biomasse végétale est une alternative durable et écologique pour les ressources fossiles. L'exploitation et la valorisation de cette biomasse sont rendues possibles grâce à la capacité naturelle des enzymes fongiques à dégrader et modifier cette biomasse. Parmi ces enzymes, les glyoxal oxydases génératrices de H2O2 (GLOX) restent un groupe peu étudié avec un seul exemple de protéine caractérisée dans la littérature à partir d’un champignon dégradant le bois.Dans cette thèse, trois GLOX, précédemment identifiées dans le génome du champignon dégradant le bois Pycnoporus cinnabarinus (PciGLOX), ont été sélectionnées, produites par voie hétérologue et caractérisées. La caractérisation a révélé des différences entre les trois PciGLOX dans la stabilité des protéines, la spécificité du substrat et l’efficacité catalytique. Les protéines PciGLOX sont produites sous leur forme inactive et leur mécanisme d'activation a été étudié. La capacité des GLOX à catalyser la réaction d'oxydation du 5-hydroxyméthylfurfural (HMF), d’intérêt industriel, a été étudiée pour la première fois dans ce travail. Le HMF a été oxydé par PciGLOX en acide 5-hydroxyméthyl-2-furancarboxylique (HMFCA) comme produit principal. Le HMFCA est difficile à produire par catalyse chimique et est utilisé dans la production de polyesters et de produits pharmaceutiques. PciGLOX ont également été capables de produire l’acide furandicarboxylique (FDCA), qui est un précurseur dans les procédés de production du bioplastique. Ce travail ouvre de nouvelles perspectives pour étudier plus en détail le rôle de GLOX dans la dégradation de la lignocellulose, et dans les applications biotechnologiques
Plant biomass is a sustainable and eco-friendly alternative for fossil fuels. The exploitation and valorisation of plant biomass is possible through biotechnological processes that rely on the natural ability of fungal enzymes to degrade and modify this biomass. Among these enzymes are H2O2-generating glyoxal oxidases (GLOX), which haven’t been extensively studied with only one example in the literature on GLOX from wood-degrading fungi. In this thesis three GLOX, previously identified in the genome of the wood-degrading fungus Pycnoporus cinnabarinus (PciGLOX), were heterologously produced and characterisation. The three PciGLOX showed differences in their stability, substrate preferences and catalytic properties. The ability of GLOX to catalyse the biotechnologically important oxidation reaction of 5-hydroxymethylfurfural (HMF) was investigated for the first time in this work. PciGLOX oxidized HMF to 5-hydroxymethyl-2-furancarboxylic acid (HMFCA), which is difficult to produce via chemical catalysis and is used in polyesters and pharmaceutical products production. PciGLOX were also able to oxidize HMF derivatives leading to the formation of the final product furandicarboxylic acid (FDCA), which is a bioplastic precursor. PciGlOX proteins are produced in their inactive form and their activation mechanism was investigated in this thesis. This work opens new prospects to investigate more the role of GLOX in plant biomass degradation and biotechnology, and the possible optimization techniques of the catalytic properties of this enzyme

Dans ce travail, nous etudions differents aspects de la dismutation (ou oxydoreduction interne) du glyoxal. Nous nous interessons tout d'abord au mecanisme de la dismutation du glyoxal en milieu aqueux basique (reaction de cannizzaro). Par utilisation de reactifs deuteres a deux valeurs de ph (14 et 9) ou les ordres en base sont respectivement 2 et 1, nous montrons que le mecanisme est le meme. Il comporte un transfert intramoleculaire d'hydrure et non pas un arrachement de proton par la base oh. La suite de nos travaux est consacree a l'etude de la dismutation lors de reactions du glyoxal sur d'autres molecules. Apres examen de la litterature, nous proposons un mecanisme general en trois etapes a partir du cas favorable des reactions du glyoxal avec les ethylenediamines: formation d'un intermediaire dihydroxyle, perte d'un groupement oh assistee par le doublet electronique de l'azote le plus nucleophile et obtention d'un ion imminium susceptible de se rearranger. Plusieurs types de molecules possedant un ou deux atomes d'azote ont ete mises en reaction avec le glyoxal: ethylene et propylenediamines, ethanolamines, urees, aminoamides et diamides. Dans chaque cas, des molecules symetriques ou non et possedant des substituants aliphatiques et/ou aromatiques ont ete utilisees et on a compare les rendements et les proportions de regioisomeres. Les resultats obtenus sont en accord avec notre mecanisme. La reaction est sous controle electronique et les produits de reaction sont conformes a la regiochimie prevue. Enfin nous montrons, par l'utilisation de composes deuteres dans le cas des reactions du glyoxal avec les ethylenediamines, que le rearrangement a lieu par depart de proton et formation d'une forme enolique tautomere de l'amide obtenu. Nous etablissons finalement une echelle de nucleophilie des molecules en nous basant sur leur reactivite vis-a-vis du glyoxal lors des reactions de dismutation

Experiments were performed to characterize the pH, gelation time, diffusion processes, material properties, adhesive properties, and the drying variables on the material and adhesive properties of Gelatin Resorcinol Dialdehyde (GR-DIAL) tissue adhesives by varying formulation. Three adhesive formulations with altered weight content of water and glyoxal (a dialdehyde) were utilized. The adhesive formulations were characterized by pH and gelation time in situ, and absorption/desorption of water in the formed resin. Thermal analysis, mechanical testing, and lap shear adhesive bond testing were utilized to characterize fresh GR-DIAL adhesive formulations and formulations dried at 370C. From the results, the diffusion processes, material and adhesive properties of the adhesive formulations were primarily affected by hydrogen bonding, chemical cross-linking, and the existence of bound water within the bulk adhesive. Formulations with increased glyoxal content had both a higher degree of cross-linking and proportion of bound water within the bulk adhesive. The increased number of chemical cross-links greatly increased the swelling resistance of the adhesives, while, the existence of bound water within the adhesive increased the resistance to drying, and plasticized the resin by depressing the resin glass transition temperature, and increased the adhesive ductility. Hydrogen bonding increased with increased gelatin content or decreased water content, resulting in increased strength and modulus of the adhesives as well as increased adhesive strength.
Master of Science

La reaction entre un synthon chiral derive du glyoxal et des composes organometalliques conduit, en plusieurs etapes, a des composes optiquement purs. La stereochimie de la reaction depend de la nature de l'organometallique utilise lors de la premiere etape, qui consiste en la substitution d'un groupe thiophenyle: inversion majoritaire avec les organocuivreux ou retention totale avec les organozinciques. Dans le cas des organozinciques, un effet stereodirecteur d'un groupe hydroxyallylique a ete mis en evidence dans un ion iminium, qui est l'intermediaire reel de la reaction

Advanced glycation end products (AGEs) irreversibly cross-link proteins with sugars and accumulate at a higher age and in diabetes, processes which can interfere with the integration of implants into the tissue. Glyoxal is a highly reactive glycating agent involved in the formation of AGEs and is known to induce apoptosis, as revealed by the upregulation of caspase-3 and fractin (caspase-3 being a key enzyme activated during the late stage of apoptosis and fractin being a caspase-cleaved actin fragment). In this study, we investigated the influence of collagen type I coating on the cytotoxic effect of glyoxal on rat calvarial osteoblastic cells and on human osteosarcoma cells (Saos-2) grown on titanium alloy, Ti6Al4V. Activation of caspase-3 and fractin was measured by counting immunohistochemically stained cells and by flow cytometry with propidium iodide (detection of the apoptosis indicating a sub-G1 peak). Our results showed an increased number of apoptotic osteoblasts after incubation with glyoxal on Ti6Al4V discs. However, the number of apoptotic cells on collagen-coated titanium was significantly smaller than on uncoated titanium after the same treatment. The present findings demonstrate that osteoblasts treated with glyoxal undergo apoptosis, whereas collagen type I coating of titanium alloys (used for implants) has an antiapoptotic function
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich

The aim of this diploma work was to execute surface treatment of non-ionic cellulose ethers EHEC that AkzoNobel provides under the tradename Bermocoll®. In order for the cellulose ether to dissolve without forming lumps, some surface treatment is commonly required. In this work glyoxal has been used for this surface modification using two different lab-scale methods that in different ways mimic the full scale production. It is an everlasting challenge for the chemical industry to reduce the consumption of chemicals and also reduce energy consumption in the production. Therefore one objective of this work was to gain fundamental understanding about the glyoxal reaction with EHEC in terms of required equivalents glyoxal, reaction temperature and reaction time. Another aim of this work was to compare the two lab scale methods with regards to their predictivity and reproducibility of results.One method is called the dry method in which a water solution of glyoxal was added to dry, non-glyoxal treated EHEC at varying temperature during heavy agitation. The second method is called the acetone method where the EHEC and glyoxal were first suspended in acetone at room temperature, and then heated at different temperature for various time periods. The parameters in the experiments made was chosen using a design of experiments (DoE) approach in order to gain as much information as possible from a few experiments and also facilitating a statistical analysis of the results.This diploma work indicates that the acetone method have a better reproducibility and would be the better choice when investigating various parameters for the reaction. On the other hand the dry method might be better suited for the further analysis of temperature dependence of the glyoxal reaction with cellulose ethers. The most important factor for the reaction was the amount of glyoxal used, whereas it is indicated that the reaction temperature had a minor effect on the reaction yields.
Program: Högskoleingenjörsexamen i Kemiingenjör – tillämpad bioteknik

Le glyoxal presente un grand interet synthetique surtout si l'on peut engager ses deux fonctions dans des reactions regio et stereoselectives. Ce travail concerne l'utilisation de derives dissymetriques du glyoxal en synthese asymetrique. La premiere partie de ce travail decrit des essais de dissymetrisation, faites a partir de glyoxal ou de glyoxal masque, qui n'ont pas donne satisfaction. La deuxieme partie decrit d'abord la preparation de divers monoacetals chiraux du glyoxal, obtenus par acetalisation des monothioacetals correspondants suivie de l'hydrolyse de la fonction thioacetal. La diastereoselectivite de l'addition d'organometalliques sur la fonction aldehyde adjacente a l'acetal est assez modeste, quel que soit le diol dont est issu l'acetal. La troisieme partie decrit la preparation de plusieurs acetals hydrazones chiraux par transacetalisation de dimethylacetalhydrazone. Puis on etude la diastereoselectivite de l'addition d'organolithiens sur la fonction hydrazone, qui est assez bonne en general et peut etre excellente si on choisit judicieusement le diol dont est issu l'acetal chiral. Nous proposons un mecanisme (chelation du lithium) en accord avec les resultats obtenus. La coupure non epimerisante de la liaison n-n peut se faire dans des conditions d'hydrogenation bien controlees. Apres oxydation on obtient l'aminoacide correspondant avec une bonne purete optique. La quatrieme partie decrit l'obtention des diols 1,2 et 1,3 symetriques ou non, qui ont ete utilises et dont la plupart ne sont pas decrits dans la litterature. L'utilisation de derives chiraux du glyoxal, appliquee aux acetals hydrazones chiraux, constitue donc une nouvelle methode de synthese d'aminoacides optiquement actifs

Advanced glycation end products (AGEs) irreversibly cross-link proteins with sugars and accumulate at a higher age and in diabetes, processes which can interfere with the integration of implants into the tissue. Glyoxal is a highly reactive glycating agent involved in the formation of AGEs and is known to induce apoptosis, as revealed by the upregulation of caspase-3 and fractin (caspase-3 being a key enzyme activated during the late stage of apoptosis and fractin being a caspase-cleaved actin fragment). In this study, we investigated the influence of collagen type I coating on the cytotoxic effect of glyoxal on rat calvarial osteoblastic cells and on human osteosarcoma cells (Saos-2) grown on titanium alloy, Ti6Al4V. Activation of caspase-3 and fractin was measured by counting immunohistochemically stained cells and by flow cytometry with propidium iodide (detection of the apoptosis indicating a sub-G1 peak). Our results showed an increased number of apoptotic osteoblasts after incubation with glyoxal on Ti6Al4V discs. However, the number of apoptotic cells on collagen-coated titanium was significantly smaller than on uncoated titanium after the same treatment. The present findings demonstrate that osteoblasts treated with glyoxal undergo apoptosis, whereas collagen type I coating of titanium alloys (used for implants) has an antiapoptotic function.
Dieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.

Le diabète est associé à divers types de complications au niveau vasculaire affectant la micro et la macro-vasculature, ce qui contribue à l'augmentation de l'incidence d'infarctus du myocarde, d'ACV, de néphropathie et de rétinopathie. Parmi les mécanismes expliquant l'apparition de ces complications, la glycation des protéines joue un rôle important. En effet, il est connu que la glycation des protéines de la matrice extracellulaire (élastine, collagène) affecte les propriétés mécaniques des tissus constitués de celles-ci. Nous pensons que la glycation des protéines du cytosquelette peut également affecter les propriétés mécaniques de cellules présentes au niveau de la vasculature, telles que la cellule du muscle lisse vasculaire ou la cellule endothéliale, et ainsi affecter les fonctions cellulaires dépendantes d'une réorganisation du cytosquelette, telles que la contraction cellulaire. Le glyoxal (GO) est un composé hautement réactif de la famille des oxoaldéhydes, considéré comme un puissant agent de glycation au niveau cellulaire, puisqu'il réagit rapidement avec les groupements amines des protéines de façon à former des produits de glycation avancés (PGA). L'étude présentée dans cette thèse démontre d'une part que l'exposition à cet agent de glycation entraîne une augmentation de la rigidité cellulaire ainsi qu'une augmentation de la réponse contractile cellulaire générée par la machinerie actomyosine, en réponse à l'AngII. À la lumière de ces résultats, nous proposons qu'une exposition au GO peut induire des modifications post-traductionnelles de type non-enzymatique des protéines impliquées dans la machinerie contractile et ainsi altérer la fonction contractile cellulaire. C'est pourquoi nous avons en second lieu évalué la glycation de trois protéines impliquées dans la machinerie contractile (actine, ROCK, gelsoline) par un essai basé sur la réaction d'une sonde fluorescente et perméable à la membrane cellulaire, soit le carboxyfluorescéine diacetate succinimidyl ester (CFDA-SE), avec les amines primaires des protéines. Par cet essai, nous avons observé une augmentation de la glycation de l'actine et de ROCK, de même qu'une augmentation de l'interaction entre l'actine et la GSN. Cette thèse montre également l'implication de l'activité kinase de ROCK dans l'amplification de la réponse contractile, suggérant que la glycation de ROCK pourrait moduler son activité. En conclusion, la modification des protéines cellulaires par le GO pourrait affecter leurs fonctions et propriétés mécaniques, notamment par la modulation d'importantes interactions protéine-protéine impliquées dans la contraction cellulaire et dans l'organisation du cytosquelette.

Dans cette recherche, il y a deux parties principales : la première est basée sur la résine mélamine-glyoxal sans utilisation de formaldéhyde, et sa modification pour qu’elle soit utilisée comme adhésifs pour bois. La deuxième partie concerne la préparation d'adhésifs écologiques et de polyuréthane non isocyanate (NIPU) à partir de biomasse (sucre, protéines, tanins, etc.). Des composites de bois tels que le contreplaqué et les panneaux de particules sont confectionnés pour mesurer la performance des adhésifs, et de nombreuses autres applications telles que le revêtement de surface et les mousses sont également testées. RMN-13C, Spectroscopie infrarouge à transformée de Fourier (FTIR), La spectrométrie de masse MALDI-TOF, analyse thermomécanique (TMA), La calorimétrie différentielle à balayage (DSC) et d’autres instruments permettant d'analyser les structures et les propriétés thermodynamiques des adhésifs. Les descriptions d'œuvres spécifiques sont les suivantes : 1)Une résine sans formaldéhyde à base de mélamine-glyoxal toxique a été synthétisé et un nouveau durcisseur à base d’acide ionique a été préparé. Le glutaraldéhyde a été utilisé pour synthétiser la résine mélamine-glyoxal-glutaraldéhyde (MGG ') et qui a donné des contreplaqués bonnes performances, permettant ainsi de remplacer sans doute la résine urée-formaldéhyde. En outre, le 5-hydroxyméthylfurfural, réactif chimique dérivé de la biomasse a été également utilisé pour modifier la résine MG afin d’améliorer ses performances de liaison et sa résistance à l’eau. 2)L’alcool furfurylique, issus de la biomasse, a été utilisé pour réagir avec des aldéhydes (formaldéhyde, glyoxal, glutaraldéhyde) afin de préparer des adhésifs pour le contreplaqué. La résine d’alcool furfurylique-glyoxal (FAG), où d’alcool furfurylique est réagi avec le glyoxal qui n’est pas toxique. La résine a donné de bonnes propriétés mécaniques et d’adhésion. C’est un adhésif écologique pour le collage du bois. L'effet de l’acidité des durcissements sur les performances de collage a été également étudié. Un durcisseur auto-neutralisant est synthétisé et utilisé. 3)Les protéines, les tanins sont aussi utilisés pour préparer des adhésifs à base de bois respectueux à l’environnementont, tout en améliorant les propriétés de liaison et la résistance à l'eau grâce à l'ajout des agents de réticulation tels que la résines alcool furfurylique-glyoxal, résines époxy, polyéthylèneimine, ..etc. 4)Des matériaux à base de biomasse tels que le monosaccharide, le disaccharide et la protéine de soja ont été utilisés pour synthétiser du polyuréthane non isocyanate (NIPU) biosourcés, utilisés pour le contreplaqué et les panneaux de particules ont permis d’obtenir des panneaux d’excellents performances. L'effet de plusieurs agents de réticulation sur la température de durcissement et les propriétés de liaison de NIPU a également été étudié. Les résines NIPU ont été appliqués dans les revêtements et les mousses rigides
In this research, there are two main parts works, first is based on melamine glyoxal resin without formaldehyde used, and the modification of it use for wood industry adhesives. The other part is the preparation of environmentally friendly adhesives and non-isocyanate polyurethane(NIPU) from biomass-based (sugar, protein, tannin, etc.). Wood composites such as plywood and particleboard were used for easuring the performance of adhesives, and more possibilities applications of them such as coating and foams were tried also. 13C-NMR, Fourier Transform Infra-Red (FTIR), Matrix assisted laser desorption ionization time-of-flight (MALDI-TOF), thermomechanical analysis(TMA), Differential scanning calorimetry(DSC) and other instruments help to analyze structures and thermodynamic properties of adhesives. The descriptions of specific works are as follows: 1) Synthetic melamine-glyoxal resin without use toxic formaldehyde and a suitable hardener was found for it. By the modification of glutaraldehyde, a melamine-glyoxal-glutaraldehyde(MGG') resin with good performance was prepared and used for plywood production, which can replace the industrial Urea-formaldehyde resin wood adhesive. In addition, 5-hydroxymethylfurfural as a chemical reagent derived from biomass material was also used to modify MG resin to improve its bonding performance and water resistance. 2) The biomass material furfuryl alcohol was used to react with aldehydes (formaldehyde, glyoxal, glutaraldehyde) to prepare plywood adhesives. Among them, furfuryl alcohol-glyoxal(FAG) resin, which was reacted with a non-toxic glyoxal, and with a good bonding propertiesis. It’s as an environmentally friendly adhesive for wood bongding. The effect of different acidity of curing agents on bonding performance has also been explored, a self-neutralizing hardener is synthesized and used. 3) Proteins, tannins were used to prepare bio-based environmentally friendly wood adhesives, and improved the bonding properties and water resistance by the addition of crosslinking agents (furfuryl alcohol-glyoxal resins, epoxy resins, polyethyleneimine, etc.). 4) Biomass materials such as monosaccharide and disaccharide were used to synthesize non-isocyanate polyurethane(NIPU), and used for plywood or particleboard to obtain bio-based NIPU adhesives with excellent performance. The effect of several crosslinkers on the curing temperature and bonding properties of NIPU was also studied. Research on more application about NIPU have also been conducted, for example, coating and rigid foams

Functionalizable spherical colloidal lignin particles (CLPs) represent a valuable asset for the valorization of lignin side-streams from the pulp industry. The spherical structure allows for the circumvention of the heterogeneous and poorly dispersible structure of the biopolymer. However, organic solvents and alkaline media degrade the particle structure and dissolve the polymers due to their chemical nature and solubility. The solvents will alter the aggregated polymers into irregular shapes that would correspond to inconsistent physicochemical properties. Then, the material will become unusable for advanced material applications, namely wood adhesives. In this study, a replicable process to yield pH ca. 12 stable CLPs for wood adhesives or further functionalization for other advanced material applications was developed and optimized. Lignin was functionalized with cross-linkers, glyoxal or formaldehyde, and selfassembled into spherical structures in the micro emulsification of the organic solution. The formed colloids were partially rotary evaporated to retain organic solvents within the colloidal structures, and then be cured at 73-76 °C until pH stable and further functionalized for advanced material applications. The functionalization with glyoxal was pursued further for its possibly increased reactivity and the health concerns associated with formaldehyde. The process requires the addition of glyoxal to lignin in an acidic organi cmedia at ambient temperature, and the solution to react at 64 °C. Glyoxal is likely added to the polymer structure in its hydrated and dimerized form, and its attachment to lignin should be analyzed through the behavior of glyoxal in different media. The formed colloids were rotary evaporated to an organic solvent content of 60 wt. % of the spheres to allow the occurrence of the curing reaction. These materials were finally cured by thermosetting them at 73-76 °C until pH stable. The particles can be cured with base-catalysis through the controlled addition of the base NaOH(aq). However, the mode and rate of addition of the catalyst are critically important for a nondegradative infusion of a base into solvent present ot removed particles without morphological changes. Further procedural improvement and larger batches are necessary to conduct CLP adhesive experiments.
Funktionaliserbara sfäriska kolloidala ligninpartiklar (CLP) är en värdefull tillgång för valorisering av ligninsidoströmmar från massaindustrin. Den sfäriska strukturen reducerar effekten av den heterogena och dåligt dispergerbara biopolymeren. Organiska lösningsmedel och alkaliska medier försämrar emellertid partikelstrukturen och löser upp polymererna på grund av deras kemiska natur och löslighet. Lösningsmedel kommer att resultera i att de aggregerade polymererna antar oregelbundna former vilket skulle resultera i inkonsistenta fysikalisk-kemiska egenskaper. Därigenom blir materialet oanvändbart för avancerade materialapplikationer, såsom t ex trälim. I denna studie, utvecklades och optimerades en reproducerbar process för att ge pH ca. 12 stabila CLP för trälim eller ytterligare funktionalisering för andra avancerade materialapplikationer. Lignin funktionaliserades med tvärbindare, glyoxal eller formaldehyd och självorganiserades till sfäriskas trukturer genom mikroemulgering av organfasen. De bildade kolloiderna indunstades delvis roterande för att bibehålla det organiska lösningsmedlet i de kolloidala strukturerna och härdades sedan vid 73-76 ° C tills pH-stabilitet och funktionaliserades ytterligare för avancerade materialapplikationer. Funktionaliseringen med glyoxal utfördes också för att reaktiviteteten och begränsa de hälsoproblem som är förknippade med formaldehyd. Förfarandet kräver tillsats av glyoxal till lignin i ett surt organiskt medium vid rumstemperatur för att sedan reageras vid 64 ° C. Glyoxal i dess hydratiserade och dimeriserade form adderas sannolikt till polymeren, och dess kemiska inbindning till lignin kan analyseras genom att undersöka glyoxal uppförande i olika medier. De bildade kolloiderna indunstades till ett organiskt lösningsmedelsinnehåll av 60 viktprocent för att möjliggöra härdning. Dessa material härdades slutligen genom värmehärdning vid 73-76 ° C tills pH var stabilt. Partiklarna kan härdas med baskatalys genom kontrollerad tillsats av basen NaOH (aq). Emellerti där sättet och tillsatshastigheten för katalysatorn kritiskt viktigt. Ytterligare processförbättringar och större satser är nödvändiga för att genomföra CLP-limexperiment.

Bien que maints brevets decrivent la reaction d'obtention du glyoxal a partir de l'ethylene glycol en presence de catalyseurs a l'argent, on constate cependant que de nombreux points fondamentaux restent a eclaircir. Aussi, apres analyse des brevets publies et mise au point de notre appareillage et des conditions de reaction, avons nous cherche a decrire le mecanisme de la reaction en proposant une expression cinetique rendant compte de nos resultats experimentaux ainsi qu'une modelisation du reacteur utilise. L'action du diethylphosphite, compose injecte en phase gazeuse a quelques ppm, a egalement ete etudiee pour interpreter son action de promoteur sur la selectivite. Les modifications de morphologie du catalyseur intervenant en cours de reaction en presence ou non de diethylphosphite ont ete caracterisees en employant des techniques d'absorption atomique, spectroscopie auger, sims ainsi que les microscopies a balayage et a effet tunnel. De plus, de nombreux catalyseurs supportes monometalliques et bimetalliques ont ete prepares et caracterises afin d'obtenir des rendements en glyoxal plus eleves que ceux decrits dans la litterature

Glyoxal and other dicarbonyl compounds have been detected in tropospheric air in rural areas and polluted urban areas. Accurate absorption cross-sections for the near UV spectrum of glyoxal are important since the value of this chemical characteristic is necessary for calculating effective quantum yields and ultimately the rates and products of photolysis. In this study, comprehensive measurements were made for the 2 80 nm gas-phase absorption spectrum of glyoxal. In addition, molecular orbital calculations were carried out and the results used as a guide to identify the vibration responsible for the weak structure features of the continuous area of the glyoxal spectrum. That vibration was found to be an ag CO stretch vibration mode of the glyoxal molecule.

Cette étude a pour objectif la mise en évidence, chez les basidiomycetes du genre Pycnoporus, de nouvelles oxydo-réductases impliquées dasn la dégradation de la biomasse végétale: de l'expression des gènes aux applications biotechnologiques. Les champs d'application visés concernent essentiellement le domaine de la chimie verte, dans le cadre du projet européen BIORENEW. Le travail s'est articulé autour de trois axes principaux. Le premier a concerné l'exploration de la biodiversité naturelle en particulier tropicale, pour la sélection de souches productrices de nouvelles laccases de haut potentiel d'oxydo-réduction. Le gène codant pour la laccase Lac1 chez Pycnoporus a été utilisé comme marqueur moléculaire d'identification et de relation phylogénie-fonction, mettant en évidence une distribution des souches fortement corrélée avec leur écozone. Le deuxième axe a porté sur l'isolement de trois nouvelles laccases issues de P.sanguineus et P. coccineus qui exhibent des caractéristiques biochimiques complémentaires: haute thermostabilité, résistance aux solvants, au pH, constantes catalytiques et potentiels rédox élevés. Ces enzymes constituent de bons modèles pour des applications en biotechnologies blanches:décoloration de colorants polyphénoliques, oxydation de composés modèles de type lignine non-phénolique, oligomérisation de flavonoides naturels adaptés aux applications cosmétiques et pharmaceutiques. Enfin, dans le cadre de l'annotation du génome des souches monocaryotiques P. cinnabarinus BRFM 137 et P; sanguineus BRFM 1264, dont le séquençage a été réalisé par notre Unité, un regard tout à fait nouveau est porté sur le système lignolytique du genre Pycnoporus, longtemps décrit comme produisant que de la laccase comme enzyme du système lignolytique. Pour la première fois, nous avons montré la présence de gènes codant pour tout l'arsenal enzymatique de dégradation des lignines, c'est à dire plusieurs laccases mais surtout de nombreuses peroxydases et des enzymes auxilliaires génératrices d'H2O2 comme les glyoxal oxydases. Ces nouvelles enzymes ont été caractérisées in silico. Pour la première fois également, la sécrétion effective de peroxydases, de glyoxal oxydases et d'autres FOLymes dans nos conditions de culture a également pu être démontrée par analyse protéomique
The purpose of this work was to prospect, in the genus Pycnoporus, for new oxido-reductases involved in the degradation of lignocellulosic biomass: from gene expression to biotechnological applications. This research was conducted in the framework of green chemistry applications according to BIORENEW European Project. The study was divided in three main research axes. Firstly, the exploration of natural biodiversity, especially tropical biodiversity, for the selection of new high redox potential-laccase producing strains. These strains were repositionned in a context of phylogenomic/function through the lac1 gene. Molecular clustering based on lac1 sequences enabled the distribution of P. sanguineus and P. coccineus through four distinct, well supported clades and subclades. This distribution was highly correlated with ecozones. The second part of the work deals with the biochemical and molecular characterization of three novel laccases from P. coccineus and P. sanguineus, and their applicability on natural or model phenolic substrates. The three laccases showed complementary biochemical features: high thermo- and pH stability, high catalytic efficiency and resistance to organic solvents. The three novel laccases proved to be suitable models for white biotechnology processes: polyphenolic dye decolourization, non-phenolic lignin model compound oxidation, and synthesis of new oligomers from natural flavonoids suitable for cosmetic or pharmaceutical applications. Finally, annotation of genomic data from the monocaryotic strains P. cinnabarinus BRFM 137 and P. sanguineus BRFM 1264 (genomes sequenced by the UMR1163 BCF ) was performed for lignolytic enzymes. For the first time, new oxidases (peroxidases, glyoxal oxidases and other FOLymes) were evidenced in Pycnoporus and in silico characterized. Moreover, the active secretion of several of these enzymes has been demonstrated in our culture conditions by 1D-proteomic analysis

Understanding future climate requires observations of trace species that can significantly influence the chemical or radiative properties of the atmosphere. The development of optical instruments, utilizing laser-systems as high-resolution light sources, for making in situ observations of trace species from either airborne- or ground-based platforms and results from field campaigns are presented. Glyoxal, the smallest \(\alpha\)-dicarbonyl, is a common product during the oxidation of volatile organic compounds. An instrument using the technique of laser-induced phosphorescence was developed to measure glyoxal at the part per trillion by volume (pptv) level from a tower in a forest canopy. The instrument was deployed as part of the Community Atmosphere-Biosphere INteractions EXperiment. The instrumental limit of detection is 3 pptv with a 1-minute acquisition time. Nearly continuous measurements of glyoxal ranging between 5 – 75 pptv were acquired throughout the campaign and vertical gradients in the forest canopy were found to be driven by elevated temperature. A sensitive instrument using laser-induced fluorescence detection was developed and deployed into remote marine environments to measure iodine monoxide (IO) where the mixing ratio is on the order of 1 – 5 pptv. The challenges and solutions of operating in this environment and results from the field and laboratory are discussed. Laboratory experiments show that IO can be generated from Laminaria digitata, a subtidal kelp species, in the presence of ozone. Observations of the isotopic composition of condensed and vapor water above a large summertime tropical convective system obtained by Hoxotope and ICOS instruments during the TC4 campaign are used to analyze the role of convection in the lower tropical transition layer (TTL). Regions of ice that are characteristic of either convective lofting or in situ condensation are encountered above an active deep tropical convective system. Ice is an important component of water transport models in the TTL, yet its isotopic composition is a relatively unconstrained parameter. The implications from the direct observations of the isotopic composition of ice during convection is explored with respect to transport models of water in the TTL.
Chemistry and Chemical Biology

La reaction de condensation entre le glyoxal et les diamines chirales possedant un axe de symetrie c#2 conduit au monoaminal du glyoxal correspondant. A partir de ce dernier, il est possible d'acceder a des synthons chiraux tels que les imines, hydrazones, esters alpha, beta-ethyleniques qui sont des precurseurs d'aldehydes alpha-amines, aldehydes alpha-hydroxyles, de beta-lactames ou encore d'esters d'acides 3-alkylsuccinaldehydiques

The demand for biodegradable packaging materials as an alternative to synthetic ones to reduce environmental cost has seen an increase in recent years. In addition, functionalizing the packaging film to provide specific advantages like antimicrobial properties has yet to be explored thoroughly. This study considers adding antimicrobial agents to improve the quality and safety of actively packaged fresh produce using an antimicrobial enzyme (lysozyme) immobilized on a biopolymer based packaging film (corn-zein). The developed packaging material is aimed as an active biodegradable packaging to reduce bacterial contamination on the surface of fresh organic produce, specifically tomatoes. The study uses glutaraldehyde and glyoxal as binding agents to immobilize the enzyme on the packaging film. The effect of concentration of glutaraldehyde and glyoxal on the controlled release of the enzyme was studied. Concentrations of 0.1 and 0.2 g/g lysozyme: cross linking agent had controlled release properties. However, concentrations of 0 or 0.05 are about 20-30% more effective at inactivating bacteria. Antimicrobial activity in the constructed zein films are also tested against selected pathogens (Salmonella Newport and Listeria monocytogenes). Developed zein based film is tested against inoculated tomatoes to determine the efficacy of the films in reducing the pathogen population. The inoculated tomatoes are stored at room temperature over a storage period of one week. The film was able to reduce Listeria monocytogenes population by three logs but was unable to reduce the population of Salmonella Newport.
Master of Science

Le vieillissement de la peau est la résultante du chrono et du photo-vieillissement qui partagent des changements moléculaires incluant les modifications des protéines telles que la glycation. La glycation se produit lorsque le glucose ou ses dérivés dicarbonylés, le glyoxal et de méthylglyoxal, réagissent avec les protéines conduisant à la formation de produits de glycation avancée (AGE). Le but de ces travaux est de mieux comprendre le rôle des glyoxalases dans la détoxification des dicarbonyles et dans la protection des protéines au cours du vieillissement de la peau. Afin de comprendre la régulation du systeme dans la peau humaine pendant le chono et le photo-vieillissement, des coupes de peau de donneurs jeunes et âgées ont été analysées par immunohistochimie. Le système des glyoxalase, glyoxalase 1 (GLO1) et glyoxalase 2 (Glo2), et AGE ont été localisés et détectés dans les échantillons de peau. Pour comprendre le rôle des GLO en réponse au stress dicarbonyle, des kéartinocytes HaCaT ont été soumise à un stress dicarbonyle de 24h et l'expression et les activités des glyoxalases ont été déterminées. Enfin, des études protéomiques ont été réalisées sur des cellules HaCaT surexprimant Glo2 ou inhibant GLO1 et nous sommes en train d'identifier des protéines cibles des carboxymethyllysines. Nos résultats montrent que le système des glyoxalases est présent dans la peau en particulier dans les cellules basales et est capable de protéger les protéines cellulaires contre les dommages oxydatifs. Nous espérons que notre étude peut contribuer à comprendre le rôle des glyoxalases dans la maintenance protéique qui est un élément clé de l'homéostasie cellulaire au cours du vieillissement
Skin aging is the result of intrinsic and photo-aging, due to UV exposure, that both share important molecular features including alterations of proteins such as glycation. Glycation occurs when glucose or its derivates the dicarbonyl compounds glyoxal and methylglyoxal react with proteins leading to the formation of advanced glycation endproducts (AGE). The aim of this work is to better understand the role of glyoxalases in the detoxification of dicarbonyls and in the protection of keratinocyte’s proteins during skin aging.In order to understand glyoxalases regulation in human skin during intrinsic and photo-aging, skin sections from young and old donors were analysed by immunohistochemistry. The glyoxalase system, glyoxalase 1 (Glo1) and glyoxalase 2 (Glo2), and AGE were localized and detected in the skin samples. To understand the role of glyoxalases in response to dicarbonyl stress, HaCaT keratinocytes were subjected to a 24h dicarbonyl stress and the expression and activities of glyoxalases were determined. HaCaT cells silenced for Glo1 or overexpressing Glo2 were developed and were used in proteomic studies to identify target proteins preferentially glycated by dicarbonyl compounds.Finally, proteomic studies were performed on cellular clones overexpressing Glo2 or inhibiting Glo1 and we are currently identifying proteins targets for carboxymethyllysine modifications.Our results show that glyoxalase system is present in skin especially in the basal cells and is able to protect cellular proteins against oxidative damages. We expect that our study may contribute to decipher the role of glyoxalases in protein maintenance, which is a key element of cellular homeostasis during aging

Ce travail a essentiellement porté sur la caractérisation et les propriétés rhéologiques de systèmes réticulés présentant des caractéristiques de gels faibles en vue de leur application au traitement des puits (prévention des venues d'eau, correction de profil). Deux types de polymères hydrosolubles ont été envisagés: un polysaccharide de fermentation (scléroglucane) et un polymère de synthèse (polyacrylamide) dont on a étudié la réticulation par le borax, le zirconium (IV) et le glyoxal. Diverses techniques (diffusion de la lumière, rhéologie, viscoélasticité) ont été utilisées pour la caractérisation des solutions et des gels. L'étude de l'influence de la salinité et de la température sur la cinétique de gélification, le diagramme de phases et les propriétes visqueuses et élastiques a particulièrement retenu notre intérêt en vue de l'application des systèmes étudiés à la technologie pétrolière. L'efficacité de divers systèmes a été testé par des essais en milieu poreux

L'@objectif de ce travail est la mise au point de nouveaux adhésifs destinés à la fabrication de panneaux de particules sans formaldéhyde. Cette molécule étant cancérogène, un autre aldéhyde, le diméthoxyéthanal (DME), non toxique, est utilisé pour formuler ces nouveaux adhésifs écologiques. Afin de mener à bien cette étude, deux voies de recherche principales sont établies. Dans un premier temps, la synthèse de résines est étudiée dans le but d'obtenir un produit stable et le plus réactif possible. La deuxième partie du travail consiste à utiliser des additifs en post addition, tels que des isocyanates et des latex, afin d'augmenter considérablement les performances des résines. Tout au long de l'étude, des panneaux de particules sont fabriqués et testés en accord avec les normes et dans le but de se rapprocher des impératifs industriels. Les formulations mises au point et les paramètres de mise en œuvre sont ainsi optimisés
This@ work aimes at formulating new adhesives intended for the manufacture of particleboard without formaldehyde. This molecule being carcinogenic, another aldehyde: the non-toxic dimethoxyethanal (DME), is being used to formulate these new environnement-friendly adhesives. To achieve a goal of this study, two main research approaches were taken. First, resin synthesis was studied with the aim of obtaining the most reactive, stable long-life products. The second part of the worl consists in using post-addition additives, such as isocyanates and latexes, to increase significantly the performance of the resins. Throughout the study, particleboards are made and tested, according to the relevant standards and with the goal of getting to industrial manufacturing conditions. Thus, the finalised formulations and the application parameters have been optimised

O presente estudo visou o desenvolvimento de biocompósitos a partir de matrizes poliméricas e reforços, com a maior proporção possível de componentes oriundos de fontes naturais. As resinas fenólicas são amplamente conhecidas e utilizadas devido à suas excelentes propriedades como estabilidade térmica e dimensional, resistência à chama e resistência química, porém, a matéria-prima utilizada na preparação desta resina (basicamente fenol e formaldeído) é obtida de fonte não-renovável. Assim, a substituição desses reagentes por equivalentes naturais corresponde a uma alternativa que vem ao encontro das preocupações atuais relacionadas com o meio ambiente, assim como pode ser vantajosa do ponto de vista econômico. Visando o aproveitamento de tanino e lignina, foi considerado o uso destas macromoléculas de origem vegetal como substitutas do fenol na preparação de resinas fenólicas do tipo resol: lignofenólica (lignina-fenol-formaldeído), lignina-formaldeído e taninofenólica. Além disso, o glioxal, um aldeído que pode ser obtido de fontes naturais, foi utilizado em substituição ao formaldeído em resinas glioxal-fenol do tipo resol e novolaca. As resinas preparadas foram caracterizadas usando espectroscopia na região de infravermelho (IV), ressonância magnética nuclear (1H e 13C RMN), termogravimetria (TG), calorimetria exploratória diferencial (DSC) e cromatografia de exclusão por tamanho (SEC). Estas resinas foram posteriormente utilizadas na preparação de termorrígidos, caracterizados por cromatografia gasosa inversa (IGC), e compósitos reforçados com fibras lignocelulósicas de sisal, com celulose isolada de sisal e celulose microcristalina, sendo os reforços caracterizados quanto à composição, cristalinidade, resistência à tração, IV, microscopia eletrônica de varredura (MEV), IGC, TG e DSC. Assim, compósitos com elevada proporção de materiais provenientes de fontes renováveis foram obtidos. Os compósitos foram caracterizados por várias técnicas, tais como, ensaio resistência ao impacto Izod, MEV, ensaio de absorção de água, análise térmica dinâmico-mecânica (DMTA), além de TG e DSC. Os resultados obtidos indicaram que a lignina e o tanino podem substituir com sucesso o fenol na preparação das matrizes fenólicas, sem que ocorra prejuízo para a resistência ao impacto, que corresponde a uma propriedade muito importante para compósitos. A absorção de água aumentou quando tanino e lignina estavam presentes na matriz, porém a variação observada não foi muito significativa, não inviabilizando o uso dos materiais obtidos em ambiente exposto à umidade. A utilização da fibra lignocelulósica de sisal e de celulose como agentes de reforço nas matrizes resultou na melhoria das propriedades mecânicas dos compósitos, aumentando a resistência ao impacto e a rigidez dos mesmos, relativamente ao termorrígido. Os compósitos reforçados com fibra lignocelulósica de sisal foram os que apresentaram maiores valores de resistência ao impacto, provavelmente devido ao comprimento destas fibras, o que contribui para a distribuição eficiente de tensões ao longo da matriz. Além disso, os resultados mostraram que a celulose de sisal e a microcristalina também podem ser consideradas como um bom material de reforço, pois apesar de não terem aumentado a resistência ao impacto de forma tão significativa, os compósitos reforçados com estes materiais absorveram menor quantidade de água, com relação àqueles reforçados com fibras lignocelulósicas de sisal. Entre os compósitos de matriz taninofenólica o reforçado com 50% de fibra de sisal foi o que apresentou a maior resistência ao impacto (416 J m-1), elevada rigidez e o menor módulo de perda, confirmando a boa interação na interface fibra/matriz deste compósito. O compósito lignofenólico reforçado com 30% de fibra lignocelulósica de sisal apresentou excelentes propriedades, como elevada resistência ao impacto (459 J m-1). Os parâmetros obtidos via IGC indicaram que as interações entre a matriz lignofenólica e a fibra de sisal devem ocorrer principalmente através de interações favoráveis entre os sítios ácidos e os sítios básicos destes materiais, possibilitando o estabelecimento de ligações hidrogênio na interface fibra/matriz. Adicionalmente, a presença de estruturas típicas de lignina na resina e nas fibras deve intensificar a afinidade entre ambas aumentando a \"molhabilidade\" da fibra durante a etapa de impregnação, intensificando a adesão fibra/matriz. As boas propriedades do compósito de matriz lignofenólica incentivaram o desenvolvimento de uma matriz em que o fenol foi totalmente substituído pela lignina, a matriz lignina-formaldeído. O compósito de matriz lignina-formaldeído reforçado com 40% de fibra de sisal foi o que apresentou a maior resistência ao impacto (512 J m-1) entre todos os compósitos preparados no presente estudo, sendo o mais indicado no caso de aplicações em que a resistência ao impacto seja um fator determinante. As imagens de MEV deste compósito revelaram uma excelente interação na interface fibra/matriz. Adicionalmente, o compósito de matriz lignina-formaldeído reforçado com 70% de fibra de sisal foi o compósito preparado com maior proporção de matérias-primas de fontes renováveis. Este compósito apresentou elevada resistência ao impacto (406 J m-1) e absorção de água comparável ao dos compósitos reforçados com menores proporções de fibras. Os compósitos reforçados com celulose de sisal e celulose microcristalina foram os que apresentaram o maior módulo de armazenamento e, portanto, maior rigidez, como consequência de a celulose ser um material de alta cristalinidade que pode agir como entrecruzador físico, aumentando a rigidez dos materiais. Os compósitos de matriz glioxal-fenol novolaca foram os compósitos que apresentaram a menor absorção de água, muito inferior à apresentada pelo compósito de matriz fenólica que é tradicionalmente usado. O compósito glioxal-fenol novolaca reforçado com celulose microscristalina apresentou absorção de água comparável à do termorrígido fenólico, com a vantagem de ser preparado com elevada proporção de materiais provenientes de fontes renováveis. No geral os compósitos, que foram preparados com elevada proporção de materiais obtidos de fontes renováveis, apresentaram excelentes propriedades, comparáveis ou até superiores aos materiais produzidos com matérias-primas provenientes de fontes não-renováveis. Estes compósitos apresentam potencial para diversas aplicações, como em partes internas de automóveis e aeronaves.
The present study aimed at developing biocomposites combining polymeric matrices and reinforcement agents, employing the highest possible proportion of materials obtained from natural sources. Phenolic resins are widely known and used due to their excellent properties, such as dimensional and thermal stability, flame resistance and chemical resistance. However, raw materials used in the production of phenolic resins, namely phenol and formaldehyde, are obtained on a large-scale from non-renewable sources. Hence, the replacement of these reagents by equivalent ones obtained from non-fossil sources is interesting from both the environmental and economical perspectives. In this study, lignin and tannin, two macromolecules obtained from natural sources, were employed as substitutes of phenol in the preparation of resol-type phenolic resins: lignophenolic (lignin-phenol-formaldehyde), lignin-formaldehyde and tannin-phenolic. Also, the glyoxal, an aldehyde that can be obtained from natural sources, was used as a substitute for the formaldehyde in the preparation of resol and novolac-type glyoxal-fenol resin. The resulting resins were analyzed using infrared spectroscopy (IR), nuclear magnetic resonance (1H and 13C NMR), thermogravimetry (TG), differential scanning calorimetry (DSC) and size exclusion chromatography (SEC). These resins were later used in the preparation of thermosets and composites reinforced with natural materials: lignocellulosic sisal fiber, cellulose isolated from sisal and microcrystalline cellulose. As a result, new composites with high proportion of materials obtained from renewable sources were developed. These composites were analyzed by Izod impact strength test, SEM, water absorption test, dynamic mechanical thermoanalysis (DMTA), TG and DSC. Thermosets were analyzed by all the tests applied to composites and also inverse gas chromatography (IGC). Reinforcements were analyzed by X ray diffraction, tensile strength test, scanning electron microscopy (MEV), IGC, IV, TG and DSC. Results indicated that lignin and tannin can successfully replace the phenol in the preparation of phenolic thermoset matrices, resulting in materials with equivalent properties, especially that of the impact strength, which represents an important property for a composite. The use of lignocellulosic sisal fiber and the celluloses as a reinforcement agent in the matrices resulted in composites with improved mechanical properties compared to the thermosets, including higher impact strength and higher stiffness. The composites reinforced with lignocellulosic sisal fibers presented the highest values of impact strength, probably due to the length of these fibers, which contributes to an efficient distribution of the tension along the matrix. Results also revealed that sisal and microcrystalline celluloses are good reinforcement agents. Although they led to a relatively lower impact strength increase, the composites reinforced with these celluloses absorbed less water than those reinforced with lignocellulosic sisal fibers. Among the composites of tannin-phenolic matrix, the composite reinforced with 50% of lignocellulosic sisal fibers presented the highest impact strength, the lowest loss modulus, and yet a high stiffness, confirming its good interaction in the fiber/matrix interface. The lignophenolic composite reinforced with 30% of lignocellulosic sisal fiber presented excellent properties such as a high impact strength. The parameters obtained by IGC indicated that the interactions between the lignophenolic matrix and the sisal fiber occur mainly by means of favorable interactions between the acid sites and basic sites of these materials. These interactions allow the establishment of hydrogen bonds in the fiber/matrix interface. In addition, the presence of typical structures of lignin in both resin and fibers improves the affinity between these two components, increasing the \"wettability\" of the fibers during the impregnation step and, consequently, increasing the fiber/matrix adhesion. The good properties of the lignophenolic composite encouraged the development of a matrix in which the phenol was totally replaced by lignin: the lignin-formaldehyde matrix. The lignin-formaldehyde composite reinforced with 40% of sisal fiber presented the highest impact strength compared to all other composites prepared in this study. Hence, this composite is the most suitable for applications where the impact strength is a crucial factor. The SEM images of this composite revealed an excellent interaction in the fiber/matrix interface. In addition, the lignin-formaldehyde composite reinforced with 70% of sisal fibers, which is the composite prepared with the highest proportion of natural materials, also presented excellent properties, such as high impact strength and low water absorption equivalent to that of composites reinforced with smaller proportion of fibers. The composites reinforced with sisal and microcrystalline cellulose presented the highest storage moduli and, therefore, the highest stiffness. This occurs mainly because cellulose is a material of high-crystallinity that can act as a physical cross-linker, increasing the stiffness of the materials. The composites of novolac glyoxal-phenol matrix presented the lowest water absorption. Actually, much lower than that of phenolic (phenol-formaldehyde) composite that is worldwide used. The novolac glyoxal-phenol composite reinforced with microcrystalline cellulose presented water absorption comparable to that of phenolic thermoset, with the advantage of having high proportion of materials from renewable sources in its composition. In summary, the composites prepared with high proportions of materials obtained from renewable sources, presented excellent properties, comparable or superior to those of materials derived from non-renewable sources. Results indicate that these new composites are feasible and interesting alternatives for a range of applications, including the manufacturing of automobile and aircraft internal parts.

Furosin entsteht bei der Salzsäurehydrolyse aus den Amadori-Produkten des Lysins und wird als Marker für den Fortschritt der frühen Maillard-Reaktion, zur Beurteilung von lebensmitteltechnologischen Prozessen sowie zur Berechnung des verfügbaren und des nicht verfügbaren Lysins in Lebensmitteln verwendet. Für die Nutzung von Furosin als Qualitätsparameter ist die reproduzierbare und konstante Bildung während der Salzsäurehydrolyse entscheidend. Dies wird in der Literatur jedoch kontrovers diskutiert. Im ersten Abschnitt dieser Arbeit galt es deshalb, die molaren Ausbeuten an Furosin und den weiteren Hydrolyseprodukten Lysin, Pyridosin und N[epsilon]-Carboxymethyl-lysin zu bestimmen und damit eine sichere Interpretation der Ergebnisse zu ermöglichen. Dazu wurden peptid-gebundene Amadori-Produkte des N[alpha]-Hippuryl-lysins in chromatographisch reiner Form dargestellt. Weiterhin wurden N[alpha]-Hippuryl-N[epsilon]-carboxymethyl-lysin und Pyridosin als Standard gewonnen. Bei den Hydrolyseexperimenten zeigten die Fructosyl-Amadori-Produkte ein ähnliches Verhalten. Nach Hydrolyse mit 6M Salzsäure wurden molare Ausbeuten an Furosin von 32% für Fructosyl-lysin und jeweils 34% für Lactulosyl- und Maltulosyl-lysin bestimmt. Signifikant höhere Ausbeuten an Furosin waren nach Hydrolyse mit 8M Salzsäure festzustellen, 46% für Fructosyl-lysin, 50% für Lactulosyl-lysin und 51% für Maltulosyl-lysin. Im Gegensatz zu den Fructosyl-Derivaten war die molare Ausbeute an Furosin bei Tagatosyl-lysin unabhängig von der verwendeten Salzsäurekonzentration (6 bis 8M) und wurde zu 42% bestimmt. Anhand der auf Basis der molaren Ausbeuten ermittelten Überführungsfaktoren kann nun erstmals die Lysin-Derivatisierung mittels der Analytik von Furosin sicher bestimmt werden. Das ermöglicht exakte Aussagen zum Fortschritt nichtenzymatischer Glykierungsreaktionen sowohl in Lebensmittel als auch in vivo. Aufgrund der Relevanz für biologische Systeme und für Lebensmittel wurden weiterhin Reaktionen von alpha-Dicarbonylverbindungen mit kurzkettigen Peptiden und dem Protein Insulin unter physiologischen Bedingungen (pH=7,4 und 37°C) untersucht. Bei der Reaktion von Glyoxal mit ausgewählten Tripeptiden wurde eine sehr schnelle Derivatisierung der Peptide und jeweils die gleichzeitige Bildung eines definierten Produktes festgestellt. Mittels nuklearmagnetischer Resonanzspektroskopie und massenspektroskopischer Analyse konnten die Produkte zweifelsfrei, jeweils als die am N-Terminus durch einen 2(1H)-Pyrazinon-Ring modifizierten Peptide, aufgeklärt werden. Das Hauptprodukt der Reaktion von Methylglyoxal mit dem Peptid Gly-Ala-Phe wurde ebenfalls als 2(1H)-Pyrazinon-Peptid aufgeklärt. Nach Inkubation von Insulin mit Glyoxal unter physiologischen Bedingungen in verdünnter Lösung konnte weiterhin gezeigt werden, dass die 2(1H)-Pyrazinon-Bildung ebenfalls an einem Protein erfolgt. Die identifizierten N-terminalen 2(1H)-Pyrazinone weisen charakteristische UV-Absorptions- sowie Fluoreszenz-Spektren auf. Um die Reaktivität des N-Terminus und damit die Bedeutung der 2(1H)-Pyrazinon-Bildung beurteilen zu können, wurden vergleichende Studien mit dem als Hauptreaktionspartner für alpha-Dicarbonylverbindungen angesehenen Arginin durchgeführt. Bei diesen Experimenten zeigte der N-Terminus und peptidgebundenes Arginin eine nahezu identische Reaktivität. Auf Grund dieser Ergebnisse ist fest davon auszugehen, dass es sich bei den identifizierten N-terminalen 2(1H)-Pyrazinonen um eine neue Klasse von sogenannten Advanced Glycation Endproducts (AGEs) mit Bedeutung in physiologischen Systemen und in Lebensmitteln handelt.

Peroxinitrito é um potente agente oxidante, nitrante e nucleofilico formado in vivo pela reação difusional do radical ânion superóxido com óxido nítrico, cuja produção exacerbada em situações de estresse oxidativo e nitrosativo pode resultar em danos a biomoléculas e estruturas sub-celulares. Por outro lado, vários compostos carbonílicos reativos tais como acroleína e compostos α-dicarbonílicos são descritos como citotóxicos e genotóxicos, pois reagem com biomoléculas aminadas resultando em perda de funções nativas, situação esta denominada de \"estresse carbonílico\". Dentre os metabólitos α-dicarbonílicos, altamente suscetíveis a adições nucleofílicas, destacam-se o biacetilo, derivado do metabolismo hepático de etanol e contaminante de alimentos, e metilglioxal e glioxal, ambos catabólitos de glicose, proteínas e lipídios acumulados em doenças relacionadas ao envelhecimento. Neste trabalho, observou-se que, em tampão fosfato normalmente aerado de pH próximo à neutralidade, (i) estes três compostos sofrem adição nucleofílica de peroxinitrito com constantes de velocidade de segunda ordem uma a três ordens de grandeza acima dos valores relatados para compostos monocarbonílicos (k2 ≈ 4-100 M-1s-1); (ii) os sistemas biacetilo ou metilglioxal/peroxinitrito consomem o oxigênio dissolvido com produção de acetato ou acetato e formiato, respectivamente, via radical acetila capaz de acetilar histidina, lisina e 2\'-desoxiguanosina se adicionados à mistura reacional; e (iii) o sistema glioxal/peroxinitrito gera sucessivamente radical formila e radical formilhidroperoxila, cujo desproporcionamento a formiato e gás carbônico é acompanhado da emissão de luz no infra-vermelho próximo (λmax = 1270 nm), atribuída a oxigênio molecular no estado singlete (O21Δg) (Reação de Russell). Estes estudos evidenciam que a reação de metabólitos α-dicarbonílicos com peroxinitrito gera radicais livres e embasam a hipótese de que possam contribuir para a acetilação radicalar, não-enzimática intracelular, de proteínas (epigenética) e DNA, portanto potencialmente implicadas na fisiologia e patologia do envelhecimento e desordens metabólicas, nas quais a participação de espécies reativas de oxigênio, nitrogênio e compostos carbonílicos foi relatada. Deve-se ainda notar que a descoberta de acetilação radicalar de biomoléculas por metabólitos α-dicarbonilicos e peroxinitrito prepara o caminho para a identificação de novas reações químicas de biomoléculas, não catalisadas por enzimas, que possam eventualmente revelar novos biomarcadores teciduais em doenças adquiridas e inatas.
Peroxynitrite is a strong biological oxidant, nitrating and nucleophilic agent, formed by the diffusion-controlled reaction of the superoxide anion radical with nitric oxide, whose exacerbated production in oxidative and nitrosative stress leads to chemical damage to biomolecules and sub-cellular structures. On the other hand, various reactive carbonyl compounds like acrolein and α-dicarbonyls are reportedly cytotoxic and genotoxic for their ability to react with amino biomolecules resulting in loss of native functions, a situation named \"carbonyl stress\". Among very reactive α-dicarbonyls highly prone to nucleophilic additions, we highlight biacetyl, a hepatic alcohol metabolite and food contaminant, and methylglyoxal and glyoxal, both catabolites of glucose, proteins and lipids that accumulate in ageing-related disorders. Here, we report that, in normally aerated phosphate buffer near the physiological pH, (i) these three dicarbonyls undergo nucleophilic addition of peroxynitrite whose second order rate constants are one to three orders of magnitude than those documented for monocarbonyls (k2 ≈ 4-100 M-1s-1); (ii) the biacetyl or methylglyoxal/peroxynitrite systems consume the dissolved oxygen yielding the acetate anion or acetate plus formate anion, respectively, from acetyl radical intermediate which was found to acetylate added histidine, lysine and 2\'-deoxyguanosine; and (iii) the glyoxal/peroxynitrite system ultimately generate formyl radical and formylperoxyl radical, whose dismutation to formate and carbonic oxide is accompanied by near infrared monomol emission (λmax = 1270 nm) characteristic of singlet molecular oxygen (O21Δg) (Russell reaction). Our studies strongly attest that the reaction of α-dicarbonyls with peroxynitrite release free radicals that can potentially contribute for the radical, non-enzymatic acetylation of proteins (epigenetics) and DNA bases possibly implicated in the ageing physiopathology and metabolic disorders, where participation of reactive oxygen, nitrogen and carbonyl species is well recognized. Also noteworthy is that our findings may pave the way to the discovery of novel biochemical reactions whose products can eventually be useful as biomarkers of acquired and innate maladies.

Furosin entsteht bei der Salzsäurehydrolyse aus den Amadori-Produkten des Lysins und wird als Marker für den Fortschritt der frühen Maillard-Reaktion, zur Beurteilung von lebensmitteltechnologischen Prozessen sowie zur Berechnung des verfügbaren und des nicht verfügbaren Lysins in Lebensmitteln verwendet. Für die Nutzung von Furosin als Qualitätsparameter ist die reproduzierbare und konstante Bildung während der Salzsäurehydrolyse entscheidend. Dies wird in der Literatur jedoch kontrovers diskutiert. Im ersten Abschnitt dieser Arbeit galt es deshalb, die molaren Ausbeuten an Furosin und den weiteren Hydrolyseprodukten Lysin, Pyridosin und N[epsilon]-Carboxymethyl-lysin zu bestimmen und damit eine sichere Interpretation der Ergebnisse zu ermöglichen. Dazu wurden peptid-gebundene Amadori-Produkte des N[alpha]-Hippuryl-lysins in chromatographisch reiner Form dargestellt. Weiterhin wurden N[alpha]-Hippuryl-N[epsilon]-carboxymethyl-lysin und Pyridosin als Standard gewonnen. Bei den Hydrolyseexperimenten zeigten die Fructosyl-Amadori-Produkte ein ähnliches Verhalten. Nach Hydrolyse mit 6M Salzsäure wurden molare Ausbeuten an Furosin von 32% für Fructosyl-lysin und jeweils 34% für Lactulosyl- und Maltulosyl-lysin bestimmt. Signifikant höhere Ausbeuten an Furosin waren nach Hydrolyse mit 8M Salzsäure festzustellen, 46% für Fructosyl-lysin, 50% für Lactulosyl-lysin und 51% für Maltulosyl-lysin. Im Gegensatz zu den Fructosyl-Derivaten war die molare Ausbeute an Furosin bei Tagatosyl-lysin unabhängig von der verwendeten Salzsäurekonzentration (6 bis 8M) und wurde zu 42% bestimmt. Anhand der auf Basis der molaren Ausbeuten ermittelten Überführungsfaktoren kann nun erstmals die Lysin-Derivatisierung mittels der Analytik von Furosin sicher bestimmt werden. Das ermöglicht exakte Aussagen zum Fortschritt nichtenzymatischer Glykierungsreaktionen sowohl in Lebensmittel als auch in vivo. Aufgrund der Relevanz für biologische Systeme und für Lebensmittel wurden weiterhin Reaktionen von alpha-Dicarbonylverbindungen mit kurzkettigen Peptiden und dem Protein Insulin unter physiologischen Bedingungen (pH=7,4 und 37°C) untersucht. Bei der Reaktion von Glyoxal mit ausgewählten Tripeptiden wurde eine sehr schnelle Derivatisierung der Peptide und jeweils die gleichzeitige Bildung eines definierten Produktes festgestellt. Mittels nuklearmagnetischer Resonanzspektroskopie und massenspektroskopischer Analyse konnten die Produkte zweifelsfrei, jeweils als die am N-Terminus durch einen 2(1H)-Pyrazinon-Ring modifizierten Peptide, aufgeklärt werden. Das Hauptprodukt der Reaktion von Methylglyoxal mit dem Peptid Gly-Ala-Phe wurde ebenfalls als 2(1H)-Pyrazinon-Peptid aufgeklärt. Nach Inkubation von Insulin mit Glyoxal unter physiologischen Bedingungen in verdünnter Lösung konnte weiterhin gezeigt werden, dass die 2(1H)-Pyrazinon-Bildung ebenfalls an einem Protein erfolgt. Die identifizierten N-terminalen 2(1H)-Pyrazinone weisen charakteristische UV-Absorptions- sowie Fluoreszenz-Spektren auf. Um die Reaktivität des N-Terminus und damit die Bedeutung der 2(1H)-Pyrazinon-Bildung beurteilen zu können, wurden vergleichende Studien mit dem als Hauptreaktionspartner für alpha-Dicarbonylverbindungen angesehenen Arginin durchgeführt. Bei diesen Experimenten zeigte der N-Terminus und peptidgebundenes Arginin eine nahezu identische Reaktivität. Auf Grund dieser Ergebnisse ist fest davon auszugehen, dass es sich bei den identifizierten N-terminalen 2(1H)-Pyrazinonen um eine neue Klasse von sogenannten Advanced Glycation Endproducts (AGEs) mit Bedeutung in physiologischen Systemen und in Lebensmitteln handelt.

Considerando a grande quantidade de resíduos sólidos, como a palha e o bagaço de cana, gerados devido às atividades agrícolas, procurou-se desenvolver um projeto que utilize esses resíduos para obtenção de produtos com maior valor agregado, visando atender às expectativas em termos econômicos e ambientais. O bagaço da cana é um resíduo gerado em grandes proporções no Brasil. O Estado de São Paulo instituiu uma lei que proíbe a prática da queimada, para fins de colheita, pelo fato de causar sérios problemas ambientais e danos à saúde da população das cidades produtoras de cana, tornando a palha mais um resíduo em abundância. O bagaço e agora a palha da cana são queimados em caldeiras para a geração de energia nas próprias usinas de açúcar e álcool. Os excedentes deste processo podem ser utilizados para a obtenção de produtos de maior valor agregado, desde polpas celulósicas bem como a produção de ligninas que podem ser utilizadas em resinas para fabricação de aglomerados. Para o uso integral da biomassa lignocelulósica é necessário fazer a separação de seus constituintes majoritários: celulose, hemicelulose e lignina. Neste trabalho, os processos de separação utilizados foram a polpação etanol-água e a técnica de explosão a vapor tendo a palha e o bagaço da cana como materiais de partida. As ligninas obtidas pelo processo etanol-água foram oxidadas em meio ácido sob condições diferentes para determinar as cinéticas da oxidação e as energias de ativação destas ligninas. Ligninas oxidadas apresentam fortes propriedades quelantes e podem ser aplicadas no tratamento de efluentes para remoção de metais pesados. A oxidação foi realizada em meio ácido acético utilizando o sitema catalítico Co/Mn/Br à 50, 80 e 115°C por 5 h. A energia de ativação calculada para as ligninas de bagaço e de palha apresentaram um valor de 23,4 kJ/mol e 34,2 kJ/mol, respectivamente, indicando que a lignina de palha é mais reticulada. O estudo cinético da oxidação foi avaliado por UV-Visível. Espectros de infravermelho de várias amostras de ligninas oxidadas foram submetidas à Analise por Componentes Principais (PCA). Os resultados mostraram suaves modificações na estrutura da lignina após a reação de oxidação. A lignina obtida pela técnica de explosão a vapor foi testada em resinas para fabricação de aglomerados, juntamente com resinas à base de farinha de soja e de tanino. Foram ainda fabricadas resinas utilizando o glioxal em substituição do formaldeído, que é um material tóxico. Lignina glioxilada foi adicionada à resina de farinha de soja glioxilada em substituição das resinas PF ou de isocianato (pMDI), visando uma maior utilização de material natural na resina. As formulações que continham 70 ou 80% de material natural apresentaram resultados dentro dos padrões exigidos. As resinas com 70% de material natural podem ser utilizadas em menor porporção na madeira e também podem ser utilizados tempos de prensagem menores, que são industrialmente significativos. A melhor formulação encontrada foi utilizando farinha de soja pré-cozida glioxilada (SG) com a adição de tanino e pMDI, onde as proporções de material foram SG/T/pMDI 54/16/30 (m/m) .
Considering the large amount of agricultural residues, such as straw and sugarcane bagasse, generated due to agricultural activities, we sought to develop a project that proposes the use of these residues to obtain products with higher value, to acchieve expectations in terms of cost and environment. Bagasse from sugarcane is a by-product generated in large proportions in Brazil. São Paulo State introduced a law which prohibits the practice of burning for harvesting because of causing serious environmental problems and damage to the health of the population of cities close to cane producers, making straw an other abundant residue. Sugarcane bagasse and straw are burned in boilers for generation of energy in sugar and alcohol industries. However, excess of those by-products could be used to obtain products with higher value, as cellulosic pulps with applications in cardboard packing and the application in resins for the manufacture of particleboard. For the integral use of the vegetable biomass it is necessary separate the major components: cellulose, hemicellulose and lignin, and for this purpose ethanol-water pulping and steam explosion process were used, with the sugarcane bagasse and straw. Lignins obtained by ethanol-water pulping were oxidized in acidic medium under different conditions to study kinetics of the oxidation and calculate the activation energies of these lignins. Oxidized lignins presents very strong chelating properties and could be applied in effluents treatments for heavy metals removal. The oxidation of lignins were performed using acetic acid and Co/Mn/Br catalytical system at 50, 80 and 115°C for 5 h. Activation energy (Ea) was calculated for lignins from sugarcane bagasse and straw and presented Ea of 34.4 kJ.mol-1 and 23.3 kJ.mol-1, respectively, indicating higher crosslinked formation for straw. A kinetic study of the oxidation was evaluated by UV/Visible. FTIR spectra of various samples of oxidized lignins were submitted to Principal Component Analysis (PCA). The results showed slight structure modifications in lignins after oxidation reaction. Lignin obtained by the steam explosion process was tested in resins for the manufacture of particleboards, together with resin-based soy flour and tannin. Resins were manufactured to substitute formaldehyde (a toxic material) with glyoxal. Glyoxalated lignins were added to glyoxalated soy flour in place of PF resins or the isocyanate (pMDI), aiming greater use of natural materials in resins. Adhesive resin formulations in which the total content of natural material is either 70 or 80% of the total resin solids content gave good results. The resins comprising 70% by weight of natural material can be used in a much lower proportion on wood chips and can afford pressing times fast enough to be significant under industrial panel pressing conditions. The best formulation of all the ones tried was the one based on glyoxalated precooked soy flour (SG), to which a condensed tannin was added in water solution and pMDI, where the proportions of the components SG/T/pMDI was 54/16/30 by weight.

Advances in spectroscopic measurement techniques enabling highly accurate measurements of trace gases in the atmosphere are critical for furthering our understanding of the chemical processes that impact both climate and human health. This dissertation presents the development and deployment of laser-based instruments for measuring parts per trillion (pptv) concentrations of iodine monoxide and glyoxal. Iodine, which is primarily released from oceanic sources, is highly reactive in the atmosphere. Despite its trace concentrations, iodine plays a potentially important role in ozone destruction, the catalysis of mercury deposition, and the formation of marine clouds. An in situ instrument to detect iodine monoxide (IO) using laser-induced fluorescence was developed and then validated during a deployment to the Shoals Marine Laboratory (Appledore Island, ME) in August and September 2011. Mixing ratios up to 10 pptv of IO were observed with a strong tidal dependence. The instrumental detection limit \((3\sigma)\) of 0.36 pptv in 1 minute is indicative of unprecedented sensitivity. Glyoxal, the smallest alpha-dicarbonyl, serves as an atmospheric tracer of both the oxidation of biogenic volatile organic compounds in forest environments as well as secondary organic aerosol. Modeling studies indicate that production of glyoxal on a global scale is driven primarily by biogenic emissions, specifically emissions of isoprene. However, measurements of glyoxal in environments where isoprene dominates its production are limited. An instrument to detect glyoxal in situ by laser-induced phosphorescence was developed. The 3σ limit of detection of this instrument was 3.9 pptv in 1 minute. During July and August 2009, gas-phase measurements of glyoxal were made during the Community Atmosphere-Biosphere Interactions Experiment at the PROPHET tower in an isoprene-dominated forest site in northern Michigan. Additional measurements made throughout the campaign have been used to constrain a box model using the Master Chemical Mechanism. The model over-predicts glyoxal relative to the observed mixing ratios. Theoretically predicted reaction pathways implemented in many isoprene oxidation schemes exacerbate this disagreement.
Chemistry and Chemical Biology

Sur Pt et Pt platine, la reduction de Co en milieu organique contenant de l'eau, conduit a la formation de methanol avec un rendement de 47% : co+4h**(+)+4e**(-)->ch::(3)oh. Le potentiel auquel a lieu cette reaction (-2,5 vvsag/ag**(+)10**(-2)m, est en accord aec une reduction simultanee de co et de h**(+) et une reaction entre les especes adsorbees sur l'electrode d'influence des adatomes sur les produits issus de l'oxydation du glyoxal et de l'ethylene glycol a ete etudiee en milieu hclo::(4) hclo::(4),n, h::(2)so::(4)2m et kno::(3)m de concentration de certaines especes, comme l'acide glyoxylique, varie enormement avec l'addition d'adatomes (ag, sn9 pour favoriser la formation d'acide formique par oxydation du glyoxal. L'oxydation de l'ethylene-glycol conduit esentiellement a la formation d'aldehyde ou d'acide formique sur electrode de pt

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Immobilization and stabilization of enzymes increases their potential for use in industrial scale. D-hydantoinases (dihidropirimidina amidrohidrolase EC 3.5.2.2) catalyze the hydrolysis of D-hydantoins, generating the corresponding Ncarbamoil- D-amino acid and are used in the production of D-amino acids, including Dphenylglycine and D-p-hydroxyphenylglycine.This work reports studies for immobilization and stabilization of D-hydantoinase from Vigna angularis (E.C. 3.5.2.2.). Different strategies of multipoint covalent attachment in organic supports as chitosan and agarose were used. Different protocols of immobilization were employed, being the adittion of ions during the reduction step with the NaBH4 important to protect enzyme catalytic site. The active and stabilized derivatives were used to catalyze the hydrolysis of D-phenylhydantoin. The temperature and pH enzyme profiles showed maximum enzyme activity at 60ºC and pH 10,0. The subunits of the enzyme present molecular mass aroundt 50kDa. The enzyme immobilized in glyoxyl-agarose in the presence of Zn2+ ions during the reduction step, with immobilization time of 24h, was the best derivative, being 89-fold more stable than the soluble enzyme. The analysis of amino acids showed that a 50% of lysines residue present in the enzymes was covalently linked in glyoxyl-agarose. The enzyme immobilized in epoxy-chitosan-alginate was 20-fold more stable than the soluble enzyme. All the tested immobilization protocols led to 100% of immobilization yield. Soluble enzyme and the best glyoxyl and chitosan enzyme derivatives were used to catalyze the hydrolysis of D- phenylhydantoin , and led to the production of 99% of NCarbamoil- D-Phenylglycine after 3, 9 and 15h of reaction respectively.
A imobilização e estabilização de enzimas aumentam muito o potencial de uso industrial desses catalisadores. D-hidantoinases (dihidropirimidina amidrohidrolase EC 3.5.2.2) são enzimas que catalisam a hidrólise de hidantoínas, com abertura do anel, para o correspondente N-carbamoil-D-aminoácido e são usadas na produção de Daminoácidos, incluindo D-fenilglicina e D-p-hidroxifenilglicina. Este trabalho relata os estudos desenvolvidos para a imobilização e estabilização de D-hidantoinase de Vigna angularis (3.5.2.2.). Foram abordadas diferentes estratégias de imobilização multipontual em suportes orgânicos como quitosana e agarose. Diferentes protocolos de imobilização foram empregados, sendo adição de íons durante a redução com NaBH4 importante para proteção do centro catalítico da enzima. Os derivados ativos e estabilizados foram empregados na reação de hidrólise da fenilhidantoína. O estudo de temperatura e pH de máxima atividade da enzima foi 60°C e pH 10,0. As subunidades da enzima apresentam peso molecular, com valor próximo a 50kDa. A enzima imobilizada em glioxil-agarose na presença dos íons Zn2+ durante a etapa de redução, com tempo de imobilização de 24 h foi o derivado mais estável sendo 89 vezes mais estável que a enzima solúvel. A análise de aminoácidos mostrou que aproximadamente 50% dos resíduos de lisina presentes na enzima foram covalentemente ligados no derivado de glioxil-agarose. A enzima imobilizada em quitosana-alginato-epoxilado foi 20 vezes mais estável que a enzima solúvel. Todos os procedimentos de imobilização testados levaram a 100% de rendimento de imobilização. Enzima solúvel e os melhores derivados obtidos por imobilização em glioxil e quitosana foram usados na catálise da hidrólise de fenilhidantoína, produzindo 99% de N-Carbamoil-D-fenilglicina nos tempos de 3, 9 e 15 h, respectivamente.

A technique for the elaboration of ene products derived from aldehydes and 2-methoxypropene into glyoxal monoketals 5 has been devised, and some reactions of the resulting aldehydes have been examined. This methodology should resolve some difficulties encountered in the course of an ongoing total synthesis of phyllanthocin. Key phases of the present study involve oxidation of ether 1, readily available from butyraldehyde through a novel ene-type reaction discovered in our laboratory, using mCPBA and methanol to give a cyclic ketal 2. The resulting seven-membered ring was opened by ketal exchange with 1,3-propanethiol to give thioketal diol 3. Benzylation of the diol proceeded with complete selectivity for the primary alcohol, permitting differentiation of the diol functionality. Protection of the secondary alcohol as a MOM derivative and subsequent ketal exchange with bis(trifluoroacetoxy)iodobenzene reinstated the dimethoxy ketal. Debenzylation under Birch conditions released the free primary alcohol 4, Swern oxidation of which yielded the desired aldehyde 5.* ftn*Please refer to the dissertation for diagrams.

碩士
國立中正大學
化學暨生物化學研究所
102
Nitric oxide is over-produced during inflammatory processes, and it reacts with superoxide anion to form peroxynitrite, resulting in the nitration of proteins. In this study, an immunoaffinity column was used to enrich nitrated and nitrosylated proteins in human urine prior to trypsin digestion. The peptides were analyzed on a nanoflow liquid chromatography nanospray ionization linear ion trap mass spectrometry and SEQUEST database searching to identify the sequence and modification sites. We analyzed the urine samples from five 4th stage oral cancer patientsand fourteen normal individuala. Five out of 46 nitrated proteins and 3 out of 67 nitrosylated proteins were identified in more than 2 samples. Glyoxal and methylglyoxal also react with proteins, resulting in the modifications of proteins at lysine and arginine residues. In this study, we use glyoxal and methylglyoxal to treat hemoglobin, and the sites of modifications were characterized by LTQ. We have identified 18 possible modifications, including 10 glyoxal- and 8 methylglyoxal-induced modifications. Compare to the non-diabetics, diabetes mellitus patients have significantly higher levels of glyoxal-induced modified peptides at α-Lys-16, α-Arg-92, β-Lys-17, β-Lys-66, and methylglyoxal-induced modified peptide at α-Arg-92. Research indicates that the concentrations of glyoxal and methylglyoxal in diabetes mellitus patients are higher than in non-diabetics. Glyoxal and methylglyoxal are mutagens and react with biological molecules. Glyoxal reacts with DNA to generate DNA crosslinked products, including dG-gx-dA and dG-gx-dC. In this study, we analyzed dG-gx-dA and dG-gx-dC adducts in diabetes mellitus patients by stable isotope dilution nanoflow ultra performance liquid chromatography nanospray ionization tandem mass spectromet ry (nanoUPLC-NSI/MS/MS) and compared them to non-diabetics. Using 50 g DNA, levels of dG-gxdC and dG-gx-dA are 1.83 ± 1.18 and 2.30 ± 0.57 in 10 8 normal nucleotides, respectively, in 9 diabetes mellitus patients’ leukocyte DNA, higher than those in non-diabetics.

Regenerated cellulose (RC) membranes are extensively used in medical and pharmaceutical separation processes due to their biocompatibility, low fouling tendency and solvent resistant properties. They typically possess ultrafiltration and microfiltration separation characteristics, but recently, there have been attempts to widen their pool of applications in nanofiltration processes. In this work, a novel method for preparing high performance composite RC membranes was developed. These membranes reveal molecular weight cut-offs (MWCO) of less than 250 daltons, which possibly put them ahead of all commercial RC membranes and in competition with high performance nanofiltration membranes. The membranes were prepared by acidic hydrolysis of dip-coated trimethylsilyl cellulose (TMSC) films. TMSC, with a degree of silylation (DS) of 2.8, was prepared from microcrystalline cellulose by reaction with hexamethyldisilazane under the homogeneous conditions of LiCl/DMAC solvent system. Effects of parameters, such as coating solution concentration and drying rates, were investigated. It was concluded that higher TMSC concentrations as well as higher solvent evaporation rates favor better MWCOs, mainly due to increase in the selective layer thickness. Successful cross-linking of prepared membranes with glyoxal solutions, in the presence of boric acid as a catalyst, resulted in MWCOs less than 250 daltons. The suitability of this crosslinking reaction for large scale productions was already proven in the manufacturing of durable-press fabrics. For us, the inexpensive raw materials as well as the low reaction times and temperatures were of interest. Moreover, the non-toxic nature of glyoxal is a key advantage in medical and pharmaceutical applications. The membranes prepared in this work are strong candidates for separation of small organic solutes from organic solvents streams in pharmaceutical industries. Their hydrophilicity, compared to typical nanofiltration membranes, offer high fouling resistance and higher fluxes in aqueous applications.

博士
國立中正大學
化學暨生物化學研究所
100
Glyoxal and methylglyoxal are mutagens and they react with biological molecules. Glyoxal reacts with DNA to generate DNA crosslinked products, include dG-gx-dC, dG-gx-dG, and dG-gx-dA. In this study, we use capillary liquid chromatography nanospray ionization tandem mass spectrometry (capLC-NSI/MS/MS) assay to quantify the levels of dG-gx-dC, dG-gx-dG, and dG-gx-dA in human placental DNA. We compared two enzyme hydrolysis conditions, and significantly different adduct levels were observed. Glyoxal and methylglyoxal also react with proteins, resulting in the posttranslational modifications of proteins. In this study, we use glyoxal and methylglyoxal to treat hemoglobin, and the sites of modifications were characterized by accurate mass measurement and by their MS2 and MS3 spectra obtained by LTQ. In human blood samples, we have identified 12 post-translational modifications, included 8 glyoxal- and 4 methylglyoxal-induced modifications and quantified the levels for this post-translational modifications relative to the reference peptides. Nitric oxide is over-produced during inflammatory processes, and then the formation of peroxynitrite, resulting in the nitration of proteins. In this study, we use peroxynitrite to treat hemoglobin, and the sites of modifications were characterized by accurate mass measurement and by their MS2 and MS3 spectra obtained by LTQ. We have identified nitration on three tyrosine residues, nitrosylation on one tyrosine, sulfoxide formation on all three methionine residues, sulfonic acid on all three cyseine and sulfinic acid on one cysteine. The post-translational modifications were semi-quantified in 20 smokers’ and nonsmokers’ blood samples. The results showed a statistically significant positive correlation between cigarette smoking and the relative quantification of tyrosine nitration at α-Tyr-24 and α-Tyr-42. Thus, measurement of thes PTMs in hemoglobin might be used as feasible biomarker for assessment of nitrative stress in vivo. Peroxynitrite not only reacts with tyrosine but also with tryptophan and cysteine. The nitrated proteins of human urinary proteins were enriched by immunoprecipitation with two types of anti-nitrotyrosine antibodies, fractionated by 1D SDS PAGE, and analyzed by nanoLC/MS/MS. So far, we have identified 25 nitrated and nitrosylated proteins in human urinary proteins.

碩士
東海大學
化學工程與材料工程學系
102
This study produced a porous polyvinyl alcohol foam by using polyvinyl alcohol as a reactant which reacted with formaldehyde, propionaldehyde and glyoxal, respectively. This study used two approaches to prepare the polyvinyl alcohol foam. One being Phase Separation Method. For the other approach, wheat starch was added (as a pore-forming agent). Then two methods were adopted aldehyde (as Cross-linking agent), and sulfuric acid (as catalyst to complete the acetalization). Finally, several physical analyses were performed to test the samples. This study adopted the following methods to analyze the polyvinyl foams: 1. Used the FTIR to confirm the reaction mechanism. 2. Used the DSC to analyze the glass transition temperature. 3. Used the SEM to observe the polyvinyl foam’s pore size and internal structure. 4. Used the colorimetric analysis method to analyze porosity. 5. Used the TGA to measure of the water contents of the polyvinyl foams. 6. Used the Instron to test the mechanical properties of the polyvinyl foams. The Polyvinyl Foam became harder and had better modulus which was caused by applying the propionaldehyde as a Cross-linking agent. Besides, the increase of polyvinyl alcohol concentration lead the pore diameter of polyvinyl foam shrink and the glass transition temperature increased slightly. The study was also conducted when formaldehyde is replaced by the glyoxal which adjusted in difference percentage. When glyoxal increases, the pore diameter would inversely shrink but water retention rate increases proportionally. In the mean time , the foam’s glass transition temperature increases significantly and generates a lower modulus under the wet conditions.