Submitted by Erika Demachki (erikademachki@gmail.com) on 2017-01-18T16:16:10Z No. of bitstreams: 2 Dissertação - Aline Carlos de Oliveira - 2016.pdf: 1109303 bytes, checksum: 21ef921b9600f5b6723928640fb5b53f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-01-19T10:30:52Z (GMT) No. of bitstreams: 2 Dissertação - Aline Carlos de Oliveira - 2016.pdf: 1109303 bytes, checksum: 21ef921b9600f5b6723928640fb5b53f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-01-19T10:30:52Z (GMT). No. of bitstreams: 2 Dissertação - Aline Carlos de Oliveira - 2016.pdf: 1109303 bytes, checksum: 21ef921b9600f5b6723928640fb5b53f (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-09-22 / Matricial tablets containing cashew gum polysaccharide (PGC) have been developed in order to promote mucoadhesion and prolonged release of model drugs with different water solubilities (theophylline and ketoprofen). Tablets were prepared by mixing the drug (40% m/m), diluent and PGC (10 to 50%, m/m). The control formulations were prepared by adding diluent instead polysaccharide. Tablets were prepared in a hydraulic press with 12.5 kN compaction force and 10s dwell time. Then, tablets were evaluated for their mechanical properties, drug content, drug in vitro release and mucoadhesive properties. The friability of the tablets met the limits recommended by the Brazilian Pharmacopoeia. The addition of the polysaccharide in the formulation significantly reduced the tablet hardness, which was also affected by the type of drug incorporated. The in vitro release test was carried out in acid medium (2h), followed by 0.4 M phosphate buffer pH 6.8 (6h). A more prolonged profile was achieved in ketoprofen-loaded tablets. In the case of theophylline-loaded tablets, prolonged release was less effective, even when the highest PGC concentration was used (50%, w/w). Ketoprofen-loaded tablets prepared with 30% of PGC released only 9% of the drug in acidic medium. In phosphate buffer, PGC tablets were capable to prolong ketoprofen release. Linearization of the drug release data showed that PGC tablets best fitted the Higuchi model. Mucoadhesion tests showed that the presence of polyssacharide increase tablet adhesion on the bowel membrane. The degree of mucoadhesion was dependent of the PGC concentration, it was greater when 30% (w/w) polysaccharide was added. The results suggests the use of cashew gum polysaccharide in matrix tablets for prolonged release of water-insoluble drugs and as mucoadhesive agent. / Comprimidos matriciais contendo polissacarídeo extraído da goma do cajueiro (PGC) foram desenvolvidos no intuito de promover mucoadesão e liberação prolongada de fármacos modelo com diferente solubilidade aquosa (teofilina e cetoprofeno). Os comprimidos foram preparados pela mistura de fármaco (40%, m/m), diluente e PGC, este último em concentração na faixa de 10 a 50% (m/m). Comprimidos controle, sem PGC, foram preparados pela substituição do polissacarídeo por diluente. A compactação foi realizada em prensa hidráulica com força de compressão de 12,5 KN exercida por 10 segundos. Os comprimidos foram avaliados quanto as suas propriedades mecânicas, teor e liberação in vitro dos fármacos incorporados e quanto à sua capacidade de mucoadesão ex vivo. A friabilidade dos comprimidos atendeu aos limites da Farmacopeia Brasileira. A adição do polissacarídeo na formulação diminuiu significativamente a dureza dos compactos, a qual também foi afetada pelo tipo de fármaco incorporado. O teste de liberação in vitro foi realizado em meio ácido (2h), seguido de tampão fosfato 0,4 M pH 6,8 (6h) e o maior prolongamento da liberação foi observado no caso do cetoprofeno. Não foi possível observar um prolongamento significativo na liberação da teofilina, mesmo com o aumento da concentração do polissacarídeo na formulação. Para o cetoprofeno, comprimidos contendo 30% de PGC, foram capazes de promover a liberação de apenas 9% do fármaco em meio ácido. Em pH 6,8 a liberação do cetoprofeno foi gradual. A linearização dos dados de liberação mostrou que o modelo de Higuchi foi o que melhor descreveu o comportamento das matrizes. Os testes de mucoadesão demonstraram que a presença do polissacarídeo conferiu mucoadesividade à forma farmacêutica. O grau de mucoadesão foi dependente da concentração do PGC na formulação, sendo maior quando 30% (m/m) do polissacarídeo foram adicionados à formulação. Os resultados sugerem o uso do PGC como agente prolongador da liberação de fármacos pouco solúveis em água em comprimidos matriciais, assim como demonstram seu potencial como agente mucoadesivo.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / O cÃncer à uma doenÃa que afeta cada vez mais um nÃmero maior de pessoas em todo o mundo. As terapias atuais utilizadas para o tratamento do cÃncer sÃo ainda insatisfatÃrias. Produtos naturais tÃm sido avaliados para seleÃÃo de compostos ativos, capazes de reduzir os tumores malignos de uma forma mais eficiente e com menos efeitos indesejÃveis. O polissacarÃdeo sulfatado extraÃdo da alga Champia feldmannii (Cf-PLS) foi testado para avaliaÃÃo do seu potencial antitumoral e imunoestimulante. AlÃm disso, foram feitos testes de toxicidade do fÃgado, rins e baÃo apÃs o tratamento com Cf-PLS. Os resultados demonstraram que Cf-PLS nÃo apresenta citotoxicidade in vitro, mas à capaz de reduzir o tumor Sarcoma 180 implantado em camundongos em 48,16% e 48,62% nas doses de 10 e 25mg/kg, respectivamente, e reduz 68,32% quando adiministrado juntamente com 5-Fluorouracil (5-FU). As anÃlises do fÃgado e rins revelaram que houve certa toxicidade no rim, com Ãreas de necrose tubular aguda na maior dose. Os testes da perfusÃo renal revelaram que Cf-PLS causa aumento da pressÃo de perfusÃo, resistÃncia vascular renal, ritmo de filtraÃÃo glomerular e fluxo urinÃrio, alÃm da excreÃÃo de sÃdio, cloreto e potÃssio. NÃo houve alteraÃÃes nos percentuais de transporte totais ou tubular proximais de sÃdio, cloreto ou potÃssio. Houve discreta deposiÃÃo protÃica nos glomÃrulos e tÃbulos renais. NÃo houve alteraÃÃes nas anÃlises bioquÃmicas e os testes hematolÃgicos revelaram uma leucopenia causada por 5-FU, entretanto esta foi revertida pelo tratamento com Cf-PLS. TambÃm foi demonstrado que Cf-PLS age como agente imunoestimulante e imunomodulador, aumentando a produÃÃo de anticorpos totais e especÃficos contra Cf-PLS e OVA, aumentando tambÃm a polpa branca e o nÃmero de megacariÃcitos nos baÃos dos animais tratados e induzindo a migraÃÃo de neutrÃfilos para a cavidade peritoneal de camundongos. Pode-se concluir que Cf-PLS apresenta atividade antitumoral e que isso pode estar relacionado com suas propriedades imunoestimulantes. / Cancer is a desease that occurs in a larger number of people each year. The therapies used to treat it are still not satisfatory. Natural products have been studied to select new compounds capable of reducing tumours and with fewer side effects. The sulfated polysaccharide isolated from the seaweed C. feldmannii (Cf-PLS) was investigated for its antitumor and immunostimulating properties and also for the toxicological aspects related to Cf-PLS treatment. The Cf-PLS did not show any significant in vitro cytotoxic effect, but showed a strong in vivo antitumor effect. The inhibition rates of sarcoma 180 tumor development were 48.16 % and 48.62% at the doses of 10 and 25 mg/kg, respectively, and 68.32% when treated together with 5-fluorouracil (5-FU). The histopathological analysis of liver and kidney showed that the kidneys were affected by Cf-PLS-treatment, presenting some focal areas of acute tubular necrosis at the higher dose. Cf-PLS caused considerable changes in renal physiology, as shown by an increase in parameters such as perfusion pressure, renal vascular resistance, glomerular filtration rate, urinary flow and sodium, chloride and potassium excretion. Neither enzymatic activity of alanine aminotransferase, urea nor creatinin levels were significantly altered. In hematological analyses, leucopeny was observed after 5-FU treatment, but this effect was prevented when the treatment was associated with the Cf-PLS. It was also demonstrated that Cf-PLS acts as an immunomodulatory agent, raising the production of specific antibodies, and also increasing the production of OVA-specific antibodies. In addition, it induced a discreet increase of the white pulp and nest of megakaryocytic in spleen of treated mice and the neutrophil migration to the intraperitoneal cavity of mice. In conclusion, Cf-PLS has some interesting antitumour activity that could be associated with its immunostimulanting properties.

Les biofilms ont longtemps été décrits comme des organisations évolutives de microorganismes, attachés à une surface et englués dans une matrice contenant, entre autre, des polysaccharides. En partant de ce constat BioFilm Control a souhaité cribler des microorganismes pour la production d’exopolysaccharides, en utilisant le BioFilm Ring Test® (BRT). Le principe repose sur la coincubation de microorganismes avec des particules magnétiques en microplaque. Les particules sont plus ou moins attirées par un aimant en fonction du stade d’organisation du biofilm. En se formant, il piège, dans sa matrice visqueuse, les particules qui perdent leur mobilité. Celle-ci est révélée par une aimantation qui provoque l’apparition d’un spot (pas de biofilm) ou non (biofilm). Une analyse d’images quantifie ce processus et permet de le standardiser. La démarche a consisté dans un premier temps à vérifier le comportement de microorganismes modèles producteurs de polysaccharides (bactéries et microalgues) avec le BRT. L’étude a été étendue au criblage d’une banque de lactobacilles. Les résultats inattendus ont orienté l’étude vers l’analyse du rôle exact des polysaccharides et plus généralement de l’implication des macromolécules dans la structuration du biofilm. Pour cela, la dégradation séquentielle de chaque famille macromoléculaire a été réalisée via des enzymes dépolymérisantes sur les biofilms de Leuconostoc mesenteroides et Bacillus sp. Au regard des résultats obtenus, l’utilisation du BRT a été étendue à la caractérisation qualitative et quantitative d’enzymes de dégradation de polysaccharides. / Biofilms were described for a long time as evolutionary structures elaborated by microorganisms, fixed on a surface and maintained in a polysaccharidic matrix. From this assessment, BioFilm Control chose to screen microorganisms for their capacity to produce exopolysaccharides (EPS), using theBioFilm Ring Test® (BRT). The principle is the co-incubation of magnetic particles with microbial culture on microplates. The mobility of particles depends on the stage of biofilm formation. During this formation, particles are trapped in the matrix and loose their mobility. Revelation is induced by magnet which causes a spot in the absence of biofilm. The pictures analysis quantifies this phenomenon and standardizes different results. This approach was realised, at first step, by the test of EPS-producing bacteria or microalgae with the BRT. The study was extended to the screening of a lactobacilli collection. Unexpected results guided the research toward the understanding of the role of macromolecules in biofilm structuring. To study their implication, sequential enzymatic degradation has been achieved for each macromolecular family of Leuconostoc mesenteroïdes and Bacillus sp. biofilms. Using the results, BRT was then appreciated as a suitable method to detect and quantify polysaccharide degrading enzymes.

<p>This thesis describes the synthesis of structures related to the capsular polysaccharide of <i>Neisseria meningitidis</i> serogroup A and the synthesis of analogues of mycothiol, a compound produced by <i>Mycobacterium</i> <i>tuberculosis</i>. The first part of the thesis describes the synthesis of structural elements present in the native capsular polysaccharide of <i>Neisseria</i> <i>meningitidis</i> serogroup A. In this part, an improved synthesis of 2-azido-2-deoxy-D-mannopyranose is included. The second part of the thesis describes the formation of stable C-phosphonate analogues related to the capsular polysaccharide. The last part outlines the formation of analogues of mycothiol, where the syntheses of a bicyclic analogue and a thioglycosidic analogue are described.</p>

<p>Polysaccharides are found in all forms of life and serve diverse purposes. They are enzymatically synthesised from activated monosaccharide precursors, nucleotide sugars. One such nucleotide sugar is UDP-glucuronic acid, which is formed from UDP-glucose by the UDP-glucose dehydrogenase (UGDH) enzyme. UGDH has been proposed to have a regulatory role in the biosynthesis of polysaccharides. The aim of the studies presented in this thesis was to investigate the role of UGDH in the polysaccharide biosynthesis in three different systems: human cell culture, bacterial cultures<i> </i>and growing<i> </i>plants<i>. </i>The effects of UGDH-overexpression on polysaccharide biosyntheses and, when achievable, on UDP-sugar levels, were investigated.</p><p>A mammalian UGDH was cloned from a kidney cDNA library. Transient expression of the cloned enzyme in mammalian cells led to an increased UGDH-activity. Northern blotting analyses revealed a single transcript of 2.6 kb in adult mouse tissues whereas human tissues expressed a predominant transcript of 3.2 kb and a minor transcript of 2.6 kb.</p><p>Overexpression of the bovine UGDH in mammalian cells induced increased synthesis of the glycosaminoglycans; heparan sulphate, chondroitin sulphate and hyaluronan, without changing their relative proportions. The effects on glycosaminoglycan synthesis caused by an increased demand of UDP-glucuronic acid were studied by overexpression of hyaluronan synthase (Has3), which requires UDP-glucuronic acid as substrate. Overexpression of Has3 and coexpression of Has3 and UGDH resulted in highly augmented production of hyaluronan without noticeably affecting heparan sulfate and chondroitin sulfate synthesis.</p><p>Expression of the bacterial UGDH in <i>E. coli</i> resulted in increased formation of UDP-glucuronic acid, but, unexpectedly, also to synthesis of fewer K5 polysaccharide chains. </p><p>Overexpression of UGD1, one of four <i>A. thaliana</i> UGDH genes, in <i>A. thaliana,</i> resulted in dwarfism. Analysis of the cell wall polysaccharides showed alteration in saccharide composition. Paradoxically, the UDP-sugars derived from UDP-glucuronic acid decreased in amount.</p>

Cellulose is the most abundant biopolymer on earth and the main load-bearing structure in plant cell walls. Cellulose microfibrils are laid down in a tight parallel array, surrounding plant cells like a corset. Orientation of microfibrils determines the direction of growth by directing turgor pressure to points of expansion (Somerville et al., 2004). Hence, cellulose deficient mutants usually show cell and organ swelling due to disturbed anisotropic cell expansion (reviewed in Endler and Persson, 2011). How do cellulose microfibrils gain their parallel orientation? First experiments in the 1960s suggested, that cortical microtubules aid the cellulose synthases on their way around the cell (Green, 1962; Ledbetter and Porter, 1963). This was proofed in 2006 through life cell imaging (Paredez et al., 2006). However, how this guidance was facilitated, remained unknown. Through a combinatory approach, including forward and reverse genetics together with advanced co-expression analysis, we identified pom2 as a cellulose deficient mutant. Map- based cloning revealed that the gene locus of POM2 corresponded to CELLULOSE SYNTHASE INTERACTING 1 (CSI1). Intriguingly, we previously found the CSI1 protein to interact with the putative cytosolic part of the primary cellulose synthases in a yeast-two-hybrid screen (Gu et al., 2010). Exhaustive cell biological analysis of the POM2/CSI1 protein allowed to determine its cellular function. Using spinning disc confocal microscopy, we could show that in the absence of POM2/CSI1, cellulose synthase complexes lose their microtubule-dependent trajectories in the plasma membrane. The loss of POM2/CSI1, however does not influence microtubule- dependent delivery of cellulose synthases (Bringmann et al., 2012). Consequently, POM2/CSI1 acts as a bridging protein between active cellulose synthases and cortical microtubules. This thesis summarizes three publications of the author, regarding the identification of proteins that connect cellulose synthases to the cytoskeleton. This involves the development of bioinformatics tools allowing candidate gene prediction through co-expression studies (Mutwil et al., 2009), identification of candidate genes through interaction studies (Gu et al., 2010), and determination of the cellular function of the candidate gene (Bringmann et al., 2012). / Zellulose ist das abundanteste Biopolymer der Erde und verleiht pflanzlichen Zellwänden ihre enorme Tragkraft. Mit der Reißfestigkeit von Stahl umwickeln Zellulosefibrillen pflanzliche Zellwände wie ein Korsett. Die Orientierung der Zellulosefibrillen bestimmt zugleich die Wachstumsrichtung, indem sie den Zellinnendruck (Turgor) in die entsprechende Ausdehnungsrichtung dirigiert (Somerville et al.,2004).Folglich zeigen Mutanten mit gestörter Zellulosesynthese oft geschwollene Organe und Zellen, die sich nicht mehr gerichtet ausdehnen können (zusammengefasst von Endler und Persson,2011). Wie aber erhalten die Zellulosefibrillen ihre parallele Orientierung? Erste Experimente aus den1960ern führten zur Vermutung, kortikale Mikrotubuli leiten die Zellulosesynthasen auf ringförmigen Bahnen um die Zellen herum (Green, 1962; Ledbetter and Porter, 1963). Diese Theorie wurde 2006 mit Hilfe moderner mikroskopischer Methoden bestätigt (Paredez et al., 2006). Wie jedoch dieser Leitmechanismus funktioniert, blieb bisher unentdeckt. Durch die Kombination verschiedener genetischer und bioinformatischer Methoden, konnten wir pom2 als Zellulose defiziente Mutante identifizieren. Die Ermittlung des Genlocus durch Map-based cloning zeigte, dass es sich bei POM2 um CELLULOSE SYNTHASE INTERACTING 1 (CSI1) handelt, ein Gen, dessen korrespondierendes Protein, wie vorher von uns gezeigt, mit dem zytosolischen Teil der primären Zellulosesynthasen interagiert (Gu et al., 2010). Durch ausführliche zellbiologische Charakterisierung von POM2/CSI1 konnten wir seine zelluläre Funktion entschlüsseln. Mit Hilfe konfokaler Spinning- Disc-Mikroskopie konnten wir zeigen, dass in Abwesenheit von POM2/CSI1, Zellulosesynthasen von den Mikrotubuli- Bahnen abweichen. Der ebenfalls von den Mikrotubuli abhängige Transport der Zellulosesynthasen zur Zellmembran hingegen, war nicht beeinflusst (Bringmann et al., 2012). Demzufolge ist POM2/CSI1 das gesuchte Bindeglied zwischen aktiven Zellulosesynthasen und Mikrotubuli. In dieser Dissertationsschrift werden drei Publikationen des Autors zusammengefasst, die wa ̈hrend der Arbeit an der Dissertiation entstanden sind. Sie beinhalten die Entwicklung bioinformatischer Methoden zur Ko- Expressionsanalyse, um Kandidatengene zu ermitteln (Mutwil et al., 2009), die Identifikaton des Kandidatengens POM2/CSI1 in einer Interaktionsstudie (Gu et al., 2010), sowie die Bestimmung der zellula ̈ren Funktion des korrespondieren- den Proteins POM2/CSI1 (Bringmann et al., 2012).

Polysaccharides are found in all forms of life and serve diverse purposes. They are enzymatically synthesised from activated monosaccharide precursors, nucleotide sugars. One such nucleotide sugar is UDP-glucuronic acid, which is formed from UDP-glucose by the UDP-glucose dehydrogenase (UGDH) enzyme. UGDH has been proposed to have a regulatory role in the biosynthesis of polysaccharides. The aim of the studies presented in this thesis was to investigate the role of UGDH in the polysaccharide biosynthesis in three different systems: human cell culture, bacterial cultures and growing plants. The effects of UGDH-overexpression on polysaccharide biosyntheses and, when achievable, on UDP-sugar levels, were investigated. A mammalian UGDH was cloned from a kidney cDNA library. Transient expression of the cloned enzyme in mammalian cells led to an increased UGDH-activity. Northern blotting analyses revealed a single transcript of 2.6 kb in adult mouse tissues whereas human tissues expressed a predominant transcript of 3.2 kb and a minor transcript of 2.6 kb. Overexpression of the bovine UGDH in mammalian cells induced increased synthesis of the glycosaminoglycans; heparan sulphate, chondroitin sulphate and hyaluronan, without changing their relative proportions. The effects on glycosaminoglycan synthesis caused by an increased demand of UDP-glucuronic acid were studied by overexpression of hyaluronan synthase (Has3), which requires UDP-glucuronic acid as substrate. Overexpression of Has3 and coexpression of Has3 and UGDH resulted in highly augmented production of hyaluronan without noticeably affecting heparan sulfate and chondroitin sulfate synthesis. Expression of the bacterial UGDH in E. coli resulted in increased formation of UDP-glucuronic acid, but, unexpectedly, also to synthesis of fewer K5 polysaccharide chains. Overexpression of UGD1, one of four A. thaliana UGDH genes, in A. thaliana, resulted in dwarfism. Analysis of the cell wall polysaccharides showed alteration in saccharide composition. Paradoxically, the UDP-sugars derived from UDP-glucuronic acid decreased in amount.

This thesis describes the synthesis of structures related to the capsular polysaccharide of Neisseria meningitidis serogroup A and the synthesis of analogues of mycothiol, a compound produced by Mycobacterium tuberculosis. The first part of the thesis describes the synthesis of structural elements present in the native capsular polysaccharide of Neisseria meningitidis serogroup A. In this part, an improved synthesis of 2-azido-2-deoxy-D-mannopyranose is included. The second part of the thesis describes the formation of stable C-phosphonate analogues related to the capsular polysaccharide. The last part outlines the formation of analogues of mycothiol, where the syntheses of a bicyclic analogue and a thioglycosidic analogue are described.

UDP-galactopyranose mutase (EC 5.4.99.9; UGM), the product of the glf gene, is an enzyme that catalyzes the conversion of uridine diphosphate galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf). UGM activity is found in bacteria, parasites and fungi, however is absent in higher eukaryotes. This enzyme is essential for the viability of many pathogenic organisms, such as Mycobacterium tuberculosis and Escherichia coli, due to the broad distribution of Galf in crucial structures such as the cell wall or capsular polysaccharide. Not surprisingly, galactofuranose biosynthesis has become an attractive antimicrobial target due to the absence of these sugars in higher eukaryotes. The UGM homologue, UDP-Nacetylgalactopyranose mutase (UNGM), was identified in Campylobacter jejuni 11168, encoded for by the cj1439c gene. UNGM is known to function as a bifunctional mutase, which catalyzes the reversible ring contraction between the pyranose-furanose forms of UDPgalactose (UDP-Gal) and UDP-N-acetylgalactosamine (UDP-GalNAc). UNGM is essential for the virulence of C. jejuni, due to the incorporation of UDP-N-acetylgalactofuranose into the capsular polysaccharide. We report the first structure of UNGM determined by X-ray crystallography, to a resolution of 1.9 Å. Analysis of the dimeric, holoenzyme structure of UNGM has identified that the cofactor flavin adenine dinucleotide is bound within each monomer of the enzyme. Comparative analysis with UGM homologues has confirmed the conserved active site residues involved in the binding of various substrates. Docking studies suggest that UNGM binds its natural substrates in a productive binding mode for catalysis with the flavin cofactor, which is consistent with the proposed mechanism for UNGM. The mobile loops are essential for substrate binding, and we have identified that the conserved arginine residue, Arg169, and the neighboring Arg168, function to stabilize the diphosphate region of UDP, although not concurrently. The non-conserved arginine residue, Arg168, appeared to favor the stabilization of N-acetylated sugars, which is in agreement with the enzyme’s higher binding affinity for UDP-GalNAc over UDP-Gal by a factor of 0.9. We have also identified that the active site Arg59 exists in two conformations in the structure of UNGM, with one conformation directed toward the active site. Arg59 is 2.5 to 3.0 Å from the acetamido moiety of GalNAc, which is favorable for stabilization and is believed to confer specificity for this substrate.

La coacervation complexe, une séparation de phase associative principalement induite par des interactions électrostatiques, entre la ß-lactoglobuline (BLG, protéine animale) et la gomme d'Acacia (AG, polysaccharide végétal) a été étudiée dans ce travail. La plus grande difficulté pour comprendre la coacervation complexe au niveau moléculaire entre BLG et AG révèle être la polymolécularité élevée d'AG. A partir de là, la motivation principale de cette thèse était de comprendre et contrôler les interactions entre la BLG et les fractions moléculaires d'AG, FI (~88% d'AG) et FII (~10% d'AG) en utilisant la titration calorimétrique isotherme, la diffusion statique et dynamique de lumière, la mobilité électrophorétique, la Granulo- Polarimétrie et la microscopie optique. Une énergie d'interaction plus forte, une stoechiométrie d'association plus faible et ainsi une complexation favorable ont étés montrées entre la BLG et FII en relation avec l'accessibilité et la densité de charges plus élevées de FII. Les résultats majeurs de cette étude ont ainsi montré des rôles différents des fractions de l'AG dans la coacervation complexe avec la BLG.