Grape pomace (GP) is one of the most abundant and underutilized fruit-derived wastes. GP is generated during winemaking, occupying over 60% of the total solid winery wastes. GP may cause serious environmental problems if it is not properly handled. On the other hand, it is rich in valuable compounds that are worthy of recovery. Although research has been working on GP upgrading, the utilizations are limited to producing a single product (e.g., grape seeds oil or polyphenol powders), which leads to large volumes of secondary wastes left. Therefore, the goal of this study is to develop an integrated process for the comprehensive utilization of GP by the production of multiple value-added products and evaluate its economic feasibility at a commercial scale. First, the chemical composition of different industrial GPs was analyzed to lay the foundation for the process design. Based on the analyzed chemical composition, an integrated process was developed to produce grape oil, polyphenols, and biofuels from GP. In this process, GP was extracted by hexane to produce oil, followed by aqueous ethanol solution extraction to obtain polyphenols. The solid residue rich in structural carbohydrates was then pretreated by alkali to partially remove lignin and enzymatically hydrolyzed to produce monomer sugars. The produced sugars were used as feedstock to produce acetone, butanol and ethanol (ABE) through anaerobic fermentation. Under the optimized conditions, the process was able to produce 71.9 g crude oil, 322.8 g crude polyphenols (equivalent to 72.6 g gallic acid), and 20.7 g ABE from 1 kg dry GP. Besides the valuable products, the process co-generated a large amount (50% of input GP biomass) of secondary waste, which is rich in lignin. Therefore, we further converted the secondary waste to biochars and evaluated their potential application in water purification by removing lead (Pb) from contaminated water. Based on the results, the produced biochar showed a high Pb adsorption ability (134 mg/g), with 66.5% of lead removal achieved within the first 30 min. Experimental and modeling results indicated that both physisorption and chemisorption mechanisms were involved in the Pb adsorption of the biochar. Finally, techno-economic analysis was conducted to evaluate the economic feasibility of the integrated processing of GP into oil, polyphenols, and biochar at an industrial scale. The results showed that compared with generating of single product or dual products, the integrated process aiming to produce multiple products had the best economic performance with the net present value (NPV), internal rate of return (IRR), and payback period of $135.0 million, 47.5%, and 1.8 years, respectively. Sensitivity analysis showed that plant capacity and polyphenol selling price had major impacts on process economics. Therefore, a suggestion for implementing this integrated process is to invest more in the polyphenol production and purification process to generate high-quality polyphenols with a high selling price and running the plant with a large capacity. Overall, we explored a novel integrated process that aims to produce multiple value-added products to increase the economic gain for the wine industry, and at the same time, potentially reduce the environmental burdens caused by GP disposal. / Doctor of Philosophy / During wine making, a large amount of solid waste is generated, and the major one is called grape pomace (GP). GP is mainly consisted of grape skins, seeds, and some stems. Normally, GP is discarded as waste; however, if it is not handled properly, GP may cause serious damages to the environment such as contaminating soil and stream water. On the other hand, GP has valuable compounds that could be recovered for other applications. Previous researchers used GP to produce a single product, which still leads to a large amount of components not used. Therefore, the aim of the current study is to design a process to comprehensively utilize GP to produce multiple value-added products. The developed process can produce grape seed oil, polyphenols, and biofuels from GP. The solid residue generated from the designed process was further converted into biochar, which can be used as an excellent adsorbent to remove lead (Pb) from contaminated water. Based on the economic model results, the developed process to convert GP into grape seed oil, polyphenols, and biochar could be a promising investment at an industrial scale. Generally speaking, various valuable products were obtained from low value GP waste, which could not only reduce the potential environmental problems caused by waste disposal, but also provide different value-added products for food, pharmacy, chemical, and energy industries.

La vescaline est une substance polyphénolique appartenant à la famille des ellagitannins C-arylglucosidiques qui inhibe in vitro la topoisomérase 2, une enzyme ciblée par les chimiothérapies utilisées contre le cancer. Jusqu’à présent, seuls des ellagitannins porteurs d’unités HHBP ont été obtenus par synthèse totale. La synthèse d’ellagitannins C-arylglucosidiques comme la vescaline, présentant un motif unique de type NHTP relié par une liaison C-arylglucosidique à une unité glucose sous forme ouverte, constitue le challenge de cette thèse. Le premier objectif a été de développer une méthodologie de couplage biarylique intramoléculaire transposable à la construction de l’unité NHTP de la vescaline. Nous avons ainsi mis au point la première méthode de couplage biarylique biomimétique, par action de l’o-chloranil, se déroulant uniquement sur des substrats poly-hydroxylés possédant un groupement méthoxyle en position anomérique du sucre. Moins restrictive, l’utilisation de complexes de cuivre-amine a permis elle aussi de réaliser ce couplage. L’étape suivante de formation biomimétique de la liaison C-arylglucosidique a ensuite été étudiée et le composé attendu a pu être obtenu avec un rendement de 25%, après optimisation des conditions, par réaction dans une solution de tampon phosphate à pH = 7.5. Les deux dernières étapes de la synthèse (galloylation sélective et formation de l’unité NHTP) ne sont pas encore réalisées mais, à ce stade, une seule étape de déprotection peut permettre d’obtenir un premier ellagitannin C-arylglucosidique naturel, l’épi-punicacortéine A 5-O-dégalloylée. / Vescalin is a polyphenol of the C-arylglucosidic ellagitannin subclass that fully inhibits topoisomerase II, an enzyme target in anticancer therapy. Up to date, most synthetic efforts have been directed towards HHBP-bearing ellagitannins. The development of a synthetic way to construct ellagitannins of the C-arylglucosidic subclass, bearing an atropoisomeric NHTP unit linked to an open-chain glucose by a C-arylglucosidic bond, as exemplified by vescalin, is the challenge of this thesis. The first objective was to establish an intramolecular biaryl coupling methodology that could be applied to the construction of vescalin’s NHTP unit. In this context, we developed the first example of biomimetic o-chloranil-mediated oxidative biaryl coupling carried out on a poly-hydroxylated substrate containing a methoxy group at the anomeric position of the sugar. Difficulties in the subsequent deprotection of the methoxy group led us to use an alternative method in which copper-amine complexes allow for the coupling of substrates bearing readily removable anomeric protecting groups. The latter methodology led to the elaboration of HHBP-containing precursors that have been subjected to the biomimetic formation of the C-arylglucosidic bond. After screening both acidic and basic conditions, the desired C-arylglucosidic compound was obtained by carrying out the reaction in a phosphate buffer (pH 7.5) in a 25% yield. The two final steps of the synthesis (selective galloylation and NHTP construction) have not been performed yet but, at this stage, we are confident that a single deprotection step will lead to the first natural C-arylglucosidic ellagitannin, 5-O-degalloyl epi-punicacortein A.

De nos jours, il y a une volonté générale de se tourner vers une agriculture plus respectueuse de l’environnement et du consommateur se traduisant notamment par une démarche de réduction des intrants chimiques. Dans un contexte de développement durable, la recherche de produits naturels pour lutter contre les maladies et les ravageurs suscite un regain d’intérêt. Dans cette thèse, des extraits hydro-alcooliques issus de coproduits de la vigne (sarment, cep, racine) et d’essences forestières (écorce d'épicéa, nœud de pin) se sont révélés être une excellente source de polyphénols bioactifs, en particulier en stilbènes complexes. En effet, ces extraits ont démontré un large spectre d’activités contre différentes maladies végétales. En particulier, un potentiel oomycide contre le mildiou de la vigne et une capacité insecticide contre un parasite des Solanacées sont rapportés. En outre, la pertinence de l'utilisation de la « chimie verte » pour extraire les stilbènes comme méthode alternative aux solvants organiques a été mise en évidence. Les présents résultats renforcent une voie de recherche originale pour faire progresser une viticulture et une agriculture plus durables, en utilisant des produits de biocontrôle moins toxiques et biodégradables, constituant ainsi une solution possible et réaliste pour lutter contre les pathogènes des plantes. / Nowadays, is a priority to turn towards a more eco- and consumer friendly agriculture resulting in the reduction of the chemical inputs. In a context of a sustainable development, the investigation of natural products to fight against diseases and pests raised a renewed interest. In this thesis, hydroalcoholic extracts derived from grapevine (cane, wood, root) and forest species (spruce bark, pine knot) by-products have demonstrated to be a great source of bioactive polyphenols, and particularly in complex stilbenes. Indeed, these extracts have proved to confer a broad spectrum of activities against different major plant diseases. In particular, an oomycide potential against downy mildew of the vine and an insecticidal capacity against Solanaceae pest were reported. Furthermore, the relevant use of “green chemistry” to extract stilbenes as an alternative method of organic solvents has been highlighted. The present findings strengthen an original line of research to advance in a more sustainable viticulture and agriculture, using less toxic and biodegradable biocontrol products, being this a possible and realistic solution to combat plant pathogens.

Made available in DSpace on 2016-04-28T14:56:20Z (GMT). No. of bitstreams: 1 Michel Cardoso.pdf: 285596 bytes, checksum: e021aeca652bc3b1ad648b8db8f92e3c (MD5) Previous issue date: 2010-03-12 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / This work aimed to study the feasibility of using tannins from the bark of Pinus oocarpa as tannin-formaldehyde adhesive to produce OSB. For the pine-bark oocarpa were tested eight forms of extraction of tannins. After evaluation of the chemical properties of tannic extracts was decided to employ the extraction with distilled water / 5% sodium sulfite to prepare for the tannin-formaldehyde adhesive. Adhesive phenol formaldehyde and urea-formaldehyde were modified with 10% tannin Pinus oocarpa and the effect of this addition on the quality of the adhesive was evaluated. OSB panels were manufactured with tannin adhesives from Pinus oocarpa (TF), Urea-Formaldehyde (UF), Phenol-Formaldehyde (PF) of UF modified with tannin and pine tannin FF modified with pine, using two times pressing . The analysis were based on the tests of static bending, tensile strength perpendicular to surface, thickness swelling and water absorption. The average density of the panels was 0.47 g/cm3. The addition of tannic extract from the bark of Pinus oocarpa proved adequate for the manufacture of OSB having in view the results for the physical and mechanical properties. The results also showed that it is possible to manufacture panels with adhesive based on pure tannin of Pinus oocarpa, however, the panels showed low strength and low dimensional stability. / Esse trabalho teve por objetivo estudar a viabilidade de utiliza??o dos taninos da casca de Pinus oocarpa como adesivo para produ??o de pain?is OSB. Para a casca de Pinus oocarpa foram testadas oito formas de extra??o dos taninos. Ap?s a avalia??o das propriedades qu?micas dos extratos t?nicos optou-se por empregar a extra??o com ?gua destilada sob adi??o de 5 % de sulfito de s?dio para confec??o do adesivo tanino-formalde?do. Adesivo Fenol- Formalde?do e Ur?ia-Formalde?do foram modificados com 10% de tanino de Pinus oocarpa e o efeito desta adi??o sobre a qualidade do adesivo foi avaliado. Foram fabricados pain?is OSB com adesivos de tanino de Pinus oocarpa (TF), de Ur?ia-Formalde?do (UF), Fenol-Formalde?do (FF), de UF modificado com tanino de pinus e FF modificado com tanino de pinus, utilizando dois tempos de prensagem. Os pain?is foram avaliados quanto ? resist?ncia a flex?o est?tica (MOR e MOE), resist?ncia ? tra??o perpendicular, (liga??o interna-LI), densidade e estabilidade dimensional (inchamento em espessura IE e absor??o em ?gua AA). O valor m?dio da densidade dos pain?is foi de 0,47 g/cm3. A adi??o de extrato t?nico da casca de Pinus oocarpa mostrou-se adequada para a fabrica??o de pain?is OSB tendo-se em vista os resultados obtidos para as propriedades f?sico-mec?nicas. Os resultados mostraram tamb?m que ? poss?vel fabricar pain?is com adesivo puro ? base de tanino de Pinus oocarpa; entretanto, os pain?is apresentaram baixa resist?ncia mec?nica e baixa estabilidade dimensional.

In this study, glucose oxidase and polyphenol oxidase were immobilized in conducting copolymer poly(N-(4-(3-thienyl methylene)-oxycarbonylphenyl)maleimide)-co-pyrrole(P(MBThi-co-Py)). A copolymer was electrochemically synthesized by using sodium dodecyl sulfate (SDS) as supporting electrolyte and characterized by FTIR, scanning electron microscopy (SEM) and conductivity measurements. Immobilization of glucose oxidase (GOD) and polyphenol oxidase (PPO) enzymes were performed in conducting PPy and P(MBThi-co-Py) matrices by electropolymerization. Kinetic parameters, maximum reaction rate (Vmax) and Michaelis-Menten constant (Km) were determined for the enzyme electrodes by help of Lineweaver-Burk plot. Effect of temperature and pH on GOD and PPO activity was examined. Operational stability and long term stability of the enzyme electrodes were investigated. The immobilized GOD and PPO electrodes were used for determination of glucose amount in Turkish orange juices and analyzing the concentration of phenolic compounds in Turkish red wines respectively.

Electrochemical polymerization of pyrrole functionalized polystyrene (PStPy) with pyrrole was carried out in water-sodium dodecyl sulfate solvent-electrolyte couple. Characterization of the resulting copolymer was performed via Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and four probe conductivity measurements. Glucose oxidase and polyphenol oxidase enzymes were immobilized in polypyrrole (PPy) and conducting copolymer of pyrrole functionalized polystyrene with pyrrole (P(PStPy-co-Py). Resulting enzyme electrodes were characterized by kinetic parameters / Vmax and Km. Behavior of enzyme electrodes upon temperature and pH changes were investigated. Glucose oxidase electrode was used for the determination of glucose in orange juice and polyphenol oxidase electrode was used for the determination of polyphenolic compounds in red wine.

In this study, two symbiotic fungi of Southern Pine Beetle (SPB), Entomocorticium peryii and Entomocorticium sp.A were evaluated in terms of polyphenol oxidase (PPO) production. The effect of different inhibitors, inducers and assay parameters such as temperature and pH on enzyme activity were investigated and maximum PPO activity was observed at 30&deg / C, pH 8.0 and when tannic acid was used as an inducer. Copper-chelator salicyl hydroxamic acid (SHAM) and pcoumaric acid, both indicated as inhibitors of tyrosinase and catechol oxidase significantly reduced the activity. For biochemical characterization studies, the enzyme was concentrated by ultrafiltration. To determine type of the enzyme, activity staining after Native-PAGE was carried out. Type of polyphenol oxidase produced by E. peryii and E. sp.A was determined as catechol oxidase by activity staining. However higher activity was observed on hydroquinone (p-diphenol) rather than catechol (o-diphenol). The enzyme obeys Michealis-Menten kinetics with Km and Vmaxvalues being 10.72 mM hydroquinone and 59.44 U/ml for E. peryii and 8.55 mM hydroquinone and 73.72 U/ml for E. sp.A respectively..

ABSTRACT IMMOBILIZATION OF INVERTASE, POLYPHENOL OXIDASE AND GLUCOSE OXIDASE IN CONDUCTING COPOLYMERS OF THIOPHENE-CAPPED POLYTETRAHYDROFURAN AND PYRROLE B&ouml / y&uuml / kbayram, AySe Elif Ph.D., Department of Chemistry Supervisor: Prof. Dr. Levent Toppare January 2005, 123 pages Immobilization of invertase, polyphenol oxidase (PPO) and glucose oxidase (GOD) enzymes were performed in electrochemically synthesized two types of conducting copolymers. One end and two end thiophene-capped polytetrahydrofuran (TPTHF-1 and TPTHF-2) were copolymerized with pyrrole under conditions of constant potential electrolysis. The copolymers were characterized by thermal, spectroscopic and scanning electron microscopy analyses. Immobilization was carried out via entrapment of enzymes in two types of matrices during the copolymerization of pyrrole with the insulating polymers in the presence of sodium dodecyl sulphate (SDS). Kinetic parameters: Maximum reaction rate (Vmax) and Michaelis-Menten constant (Km) were determined for the enzyme electrodes. Temperature optimization, pH optimization, operational stability and shelf-life of the enzyme electrodes were investigated. Enzyme electrodes of polyphenol oxidase and glucose oxidase were used to determine the amount of their substrates in samples. Polyphenol oxidase converts mono and diphenols to quinone. Amount of phenolic compounds in two kinds of wines were determined by analyzing the quinone amount. Glucose oxidase converts &amp / #61538 / -D-glucose to D-glucono-1,5-lactone. Glucose amount was determined in two kind of factory-produced orange juices by analyzing D-glucono-1,5-lactone.

La substitution des composés carbonés d'origine fossile par des matières premières renouvelables représente un enjeu majeur et stratégique pour l'environnement, l'économie, et la santé publique. Nous nous sommes particulièrement intéressés au cas des dérivés du phénol, largement utilisés dans le domaine des matériaux synthétiques (résines, plastiques). Des travaux antérieurs ont démontré le potentiel des monomères de flavanols, sous-unités constitutives des chaines polymériques des tanins condensés comme substituts aux phénols issus de la pétrochimie, tels que le bisphénol A, l'objectif du présent travail a été de contribuer au développement d'une voie industrielle d'accès à de tels composés par dépolymérisation des tanins condensés (proanthocyanidines), une ressource végétale abondante mais actuellement peu exploitée pour ce type d'applications. Cette dépolymérisation peut être obtenue par rupture de la liaison interflavanique en milieu acide et piégeage des carbocations formés par un réactif nucléophile. Différentes stratégies et conditions ont été étudiées pour mieux comprendre le rôle de l'acide, du nucléophile, du solvant et l'impact de la température sur la réaction de dépolymérisation. Les stratégies sélectionnées, menées sur un dimère B2 pur servant de modèle et sur un extrait industriel de tanins de pépins de raisins blancs issus de marcs à l'échelle du gramme, ont été comparées en termes de rendement, de cinétique, de stabilité des produits, et de facilité de mise en œuvre et de traitement dans la perspective d'une production à plus grande échelle respectant au mieux les objectifs du développement durable et les contraintes de l'industrialisation. / The substitution of fossil carbon by renewable raw materials is a major strategic challenge for the environment, economy and public health. Our thesis focuses on the utilisation of phenol derivatives, which are widely used in the field of synthetic materials (resins, plastics). Previous studies have demonstrated the potential of plant flavanol monomers as substitutes for petrochemical phenols such as bisphenol A. The objective of this work was to contribute to the development of an industrial access to such compounds by depolymerization of condensed tannins (proanthocyanidins), an abundant plant resource currently untapped for such applications. Depolymerization can be performed by breaking the interflavan bonds in acidic medium and scavenging the released carbocations by a nucleophilic reagent. Various strategies and conditions were studied and compared to better understand the role of the acid, the nucleophile and the solvent and temperature in the depolymerisation reaction. The selected strategies, applied on a pure B2 dimer used as a model and an industrial seed tannins extract from white grape pomace at the gram scale, were compared in terms of performance, kinetics, product stability, ease of implementation and work-up in the context of a larger scale production complying with the objectives of sustainable development and the constraints of industrialization.

Les polyphénols, métabolites essentiels des végétaux, possèdent des propriétés intéressantes pour la santé : antioxydants, antimicrobiens ou anticarcinogéniques mais aussi pour la physico-chimie : amphiphiles, donneurs et accepteurs de liaisons hydrogènes, complexation avec les ions métalliques. Cette thèse a pour but d’utiliser un polyphénol : l’acide tannique (AT) afin de fonctionnaliser des matériaux sous différentes formes. Tout d’abord, l’AT a permis de structurer des fibres électrospinnées d’acide poly-lactique à l’aide de la technique de co-électrospinning-électrospraying pour la régénération de la pulpe dentaire. Ensuite, il a été possible de réaliser pour la première fois des fibres électrospinnées d’AT pur et de les réticuler ouvrant la porte à une synthèse « verte » de fibres pour la catalyse notamment. Enfin, des films multicouches à l’aide d’enzymes ont été construits permettant d’obtenir des films ayant à la fois une activité enzymatique et une électro-activité. / Polyphenols are essential metabolites in vegetal kingdom. They have interesting properties both for healthcare and physical-chemistry : they are antioxidant, antimicrobial or anticarcinogenic and they are amphiphilic, hydrogen-bond acceptors and donors and they can chelate with metallic cations. This thesis aims to use a polyphenol : tannic acid (TA) to design functionnal materials. To begin, for a dental pulp engineerig purpose, TA was used to structure electrospun fibers of poly-lactic acid by using the technics of co-electrospinning-electrospraying. Then, for the first time, it was possible to electrospin fibres from a solution of pure TA. To consider applications, it was possible to cross-link these fibres. This result paves the way for a green synthesis of electrospun membranes. To finish, multilayer enzymatic films were made with TA and alkaline phosphatase or lysozyme. Films have both enzymatic properties and electro-activity due to TA.