Orientador: Crispin Humberto Garcia Cruz / Banca: Eleni Gomes / Banca: Roberto Alves de Oliveira / Resumo: Os polímeros microbianos extracelulares (exo-biopolímeros) são moléculas que apresentam um grande potencial de aplicação nas indústrias de alimentos, farmacêuticas, petrolíferas, de cosméticos, têxteis, de papéis, tintas, produtos agrícolas entre outras. Como biorremediadores, podem ser utilizados na remoção de metais pesados em ambientes aquáticos poluídos, atuando como agentes de biossorção por meio da acumulação de nutrientes a partir do meio ambiente. O objetivo do trabalho foi a produção de exobiopolímeros a partir de três Ascomicetos endofíticos (Colletotrichum sp., Guignardia sp. e Phomopsis sp.). Foi selecionado o fungo que apresentou a maior produção desse material biológico. Tanto esse exo-biopolímero quanto a biomassa foram avaliados quanto aos seus potenciais de biossorção dos íons metálicos chumbo (Pb+2) e cádmio (Cd+2). Para a obtenção do meio de cultivo utilizado para a produção de exo-biopolímeros, foram testados o meio mínimo de sais de Vogel modificado com diferentes fontes de nitrogênio definidas e glicose como única fonte de carbono. Em seguida, foi realizado um planejamento fatorial 3(3-0) com 27 experimentos em duplicata, onde as variáveis independentes foram o tempo, temperatura de cultivo e a concentração inicial de glicose. O fungo Colletotrichum sp. apresentou a maior produção de exo-biopolímero (5,71 gL-1) utilizando a uréia como fonte de nitrogênio, em cultivo a 28 ºC, 150 rpm e glicose 6 % (p/v) em 192 horas de cultivo. A execução do planejamento fatorial demonstrou que a produção de biomassa seca foi favorecida por faixas de temperatura e concentração inicial de glicose entre 24 e 32 ºC e 35 e 75 gL-1, respectivamente. A produção de exo-biopolímero foi favorecida por faixas de temperatura e concentração... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The microbial extracellular polymer (exo-biopolymers) are molecules that have a great potential for application in the industries of food, pharmaceutical, oil, cosmetics, textiles, paper, paints, agricultural products and others. As bioremediation, can be used in the removal of heavy metals in polluted aquatic environments, acting as agents of bioremediation through the accumulation of nutrients from the environment. Objective of this study was to produce exo-biopolymer from three endophytic Ascomycota (Colletotrichum sp., Guignardia sp. and Phomopsis sp.). Then, was selected the fungus that showed the highest production of exobiopolymer. Both the exo-biopolymer and biomass were assessed for their potential of biosorption of metallic ions lead (Pb+2) and cadmium (Cd+2). To obtain the culture medium used for the production of exo-biopolymers, was tested a minimal medium salts of Vogel modified with different nitrogen sources and glucose defined as the only carbon source. Next, was performed a factorial design 3(3-0) with 27 experiments in duplicate, where the independent variables were the time and temperature of cultivation and the initial concentration of glucose. Colletotrichum sp. had the highest production of exo-biopolymer (5.71 gL-1) using urea as the nitrogen source in cultivation at 28 ºC, 150 rpm, and glucose 6% (w/v) in 192 hours of culture. The implementation of the factorial design showed that the dry biomass production was enhanced by a range of temperatures and initial concentration of glucose between 24 and 32 ºC and 35 and 75 gL-1, respectively. The production of exobiopolymer was favored by a range of temperatures and initial concentration of glucose between 24 and 36 ºC and 35 and 70 gL-1, respectively. In the experiment, the fungus Colletotrichum sp. produced 6.11 gL-1 of... (Complete abstract click electronic access below) / Mestre

Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The development of the work plan proposed for this dissertation refers to the production of xanthan gum from sugarcane juice using the bacterium Xanthomonas campestris pv. campestris NRRL B-1459. Tests were conducted in 500 mL erlenmeyers flasks and in a 2.0-L reactor, in sucrose concentration range from 15 to 40 g/L, judiciously following the pre-established experimental designs, when the process variables and its levels were defined. Firstly, two varieties of sugarcane, RB 835486 and SP 791011 were tested in different production media, aiming at selecting the variety and the fermentative media whose K values (rate of consistence) and n (behavior rate) for the fermented must represented in a better way the condition of pseudoplastic fluid. In the sequence, two factorial designs in two levels and three variables defined the appropriate composition of the medium for later investigations related to the gum biosynthesis. However, the third design appears with two variables and three levels, rendering favorable an evaluation of the optimum point through surfaces generated for each response analyzed. Above all, an increase in the scale researched until then guided the execution of tests in reactor for checking, under controlled conditions, the growth of microorganisms, decrease of sucrose concentration and the product formation, the viscosity of fermented must, and the 1% solution of gum after recovery and purification. Tests in erlenmeyer flasks were carried out in stirring table at 120 rpm and 28ºC while in the reactor, the agitation, aeration, and pH maintained their values in 800 rpm, 0.5 vvm and 7.3, respectively. In that way, results found in this study indicate that the variety of sugarcane SP 791011 has remarkable potential for the production of xanthan gum and after execution of two designs was defined too the medium of production constituted by dilute sugarcane, fortified with four components for effecting the 3rd and 4th designs. Then, the feasibility of the production process of xanthan gum was attested in reactor, reaching 15.1 g/L of gum concentration, 0.629 g/Lh of productivity, 0.579 g.g-1 of conversion from substrate to product, 21509.9 cP of viscosity at 0.75 s-1 for 1% solution of xanthan gum. Gums produced were compared to a commercial sample and presented a similar spectroscopy near infrared region. / O desenvolvimento do plano de trabalho proposto para esta dissertação refere-se à produção de goma xantana a partir de caldo de cana utilizando a bactéria Xanthomonas campestris pv. campestris NRRL B-1459. Os ensaios foram conduzidos em erlenmeyers de 500 mL e em reator de 2,0 L, numa faixa de concentração de sacarose de 15 a 40 g/L, seguindo criteriosamente planejamentos experimentais pré-estabelecidos, uma vez definidas as variáveis de processo e seus níveis. Primeiramente, duas variedades de cana-de-açúcar, RB 835486 e SP 791011, foram testadas em diferentes meios de produção visando selecionar a variedade e os meios fermentativos cujos valores de K (índice de consistência) e n (índice de comportamento) para o mosto fermentado representaram melhor a condição de fluido pseudoplástico. Na seqüência, dois planejamentos fatoriais a dois níveis e três variáveis definiram a composição adequada do meio para posteriores investigações relacionadas à biossíntese da goma. Contudo, surge o 3o planejamento com duas variáveis a três níveis, propiciando uma avaliação do ponto ótimo através das superfícies geradas para cada resposta analisada. Sobretudo, um aumento na escala pesquisada até então direcionou a execução dos ensaios em reator para verificar, sob condições controladas, o crescimento de microrganismos, o decréscimo da concentração de sacarose, a formação de produto, a viscosidade do mosto fermentado e a da solução 1% da goma após recuperação e purificação. Os testes em erlenmeyers foram realizados em mesa agitadora a 120 rpm e a 28o C enquanto no reator a agitação, a aeração e o pH mantiveram seus valores em 800 rpm, 0,5 vvm e 7,3 respectivamente. Dessa forma, os resultados encontrados nesse estudo apontam a variedade de cana SP 791011 com potencialidade notável para a produção de goma xantana e definiu-se também após execução de dois planejamentos o meio de produção constituído por caldo de cana diluído fortificado com quatro componentes para efetuar o 3o e 4o planejamentos. Logo, a viabilidade do processo de produção da goma xantana foi atestada em reator, atingindo 15,1 g/L de concentração de goma, 0,629 g/Lh de produtividade, 0,579 g.g-1 de conversão de substrato a produto, 21509,9 cP de viscosidade a 0,75 s-1 para solução 1% de goma. As gomas produzidas foram comparadas a uma amostra comercial e apresentaram espectroscopia na região do infravermelho similares. / Mestre em Engenharia Química

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2017-02-06T17:32:01Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 05 LEANDRO.pdf: 1567255 bytes, checksum: 524598baa8a0da3ae61f6dd0f193fb70 (MD5) / Made available in DSpace on 2017-02-06T17:32:01Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 05 LEANDRO.pdf: 1567255 bytes, checksum: 524598baa8a0da3ae61f6dd0f193fb70 (MD5) Previous issue date: 2012-08-22 / Introdução: A reconstrução do parênquima renal requer, em algumas situações, uma técnica cirúrgica mais complexa e delicada. O trauma renal de alto grau e as nefrectomias parciais para remoção de tumores malignos induzem a uma reconstrução renal de difícil execução e possuem um potencial elevado para causar perda completa do órgão. A utilização de substâncias auxiliares na reconstrução renal, tais como: selantes biológicos e hemostáticos têm fundamental importância para a sua preservação. OBJETIVO: Avaliar o efeito hemostático e selante da esponja do biopolímero da cana-de-açúcar em trauma renal de alto grau induzido em coelhos. MÉTODOS: Foi realizado um estudo experimental, prospectivo e pareado, com vinte e quatro unidades renais de doze coelhos da linhagem California. Após indução de trauma renal bilateral e de alto grau (lesão da via coletora), utilizamos, para a reconstrução e hemostasia do parênquima renal direito, a esponja do biopolímero da cana-de-açúcar, e à esquerda, foi utilizada a esponja de Surgifoam®. Nos dois lados, foi realizada uma sutura com fio de poliglactina 4/0 sobre as esponjas. Desta forma, cada coelho se tornou controle de si mesmo. Durante o procedimento cirúrgico, foram avaliados: o tempo cirúrgico total, o tempo para hemostasia e o grau de perda sanguínea. Os animais foram sacrificados após quatro semanas, sendo os rins retirados e encaminhados para a análise histológica. Foi avaliada a presença de fístulas, urinoma, hematomas e aderências aos órgãos adjacentes no momento da nefrectomia. A análise histológica observou o grau de inflamação e fibrose, assim como presença de necrose. RESULTADOS: Em relação ao tamanho dos rins, não houve diferença estatística entre os grupos (p=0,058), o que configura uma homogeneidade entre eles. Da mesma forma, o tempo para hemostasia foi similar (p=0,195) e a perda sanguínea (p=0,480). Por outro lado, foi notada uma maior incidência de aderências aos órgãos adjacentes no grupo do BCPA, assim como, uma maior intensidade da resposta inflamatória a corpo estranho (p=0,004). Todavia, sem influência nos resultados finais. Não foi evidenciada a ocorrência de hemorragia maciça, formação de hematomas, abscessos, fístulas ou cálculos em nenhuma das unidades renais. A presença de fibrose e necrose foi semelhante nos dois grupos do ponto de vista estatístico (p=0,500 e 0,414, respectivamente). CONCLUSÃO: A esponja do biopolímero da cana-de-açúcar demonstrou eficácia em relação ao controle hemostático e selante sobre o parênquima renal de coelhos, sem causar complicações importantes. Futuros estudos podem direcionar seu uso rotineiro na prática urológica. / Introduction: The reconstruction of the renal parenchyma requires, in some situations, a surgical technique more complex and delicate. The high grade renal trauma and partial nephrectomy for removal of malignant tumors induce a kidney reconstruction difficult to implement and have a high potential to cause complete loss of the organ. The use of auxiliary substances in renal reconstruction, such as biological sealants and hemostatic are of enormous value to their preservation. Objective: To evaluate the effect of sealant and hemostatic sponge biopolymer of sugarcane in high grade renal trauma induced in rabbits. Methods: We conducted an experimental study, prospective, matched, with twenty-four renal units of twelve rabbits from California lineage. After induction of bilateral renal trauma and high-grade lesions, it was used for the reconstruction of right renal parenchymal, the biopolymer sponge of cane sugar, and in left kidney, we used the sponge Surgifoam®. On both sides, we performed a suture with polyglactin 4/0 on the sponges. Thus, each rabbit has control of himself. During surgery, we evaluated the total surgical time, time to hemostasis and the degree of blood loss. The animals were sacrificed after four weeks, and the kidneys removed and sent for histological analysis. We evaluated the presence of fistula, urinoma, hematoma, and adhesions to adjacent organs at the time of nephrectomy. Histological analysis showed the degree of inflammation and fibrosis, as well as necrosis. Results: In relation to the size of the kidneys, there was no statistical difference between groups (p = 0.058), which is one commonality between them. Similarly, time to hemostasis was similar (p = 0.195) and blood loss (p = 0.480). Moreover, it was noted an increased incidence of adhesions to adjacent organs in BCPA group, as well as a greater inflammatory response to foreign bodies (p = 0.004). However, no influence on final results. There was no evidence the occurrence of massive bleeding, hematomas, abscesses, fistulas, or in any calculations of renal units. The presence of fibrosis and necrosis was similar in both groups a statistically (p = 0.500 and 0.414, respectively). Conclusion: The biopolymer sponge of sugarcane has shown efficacy in the hemostatic control and sealant on the renal cortex of rabbits, without causing major complications. Future studies may direct its routine use in urologic practice.

In this work nanocomposites based on chitosan or carboxymethylcellulose, and different kinds of clays were prepared using polyethyleneglycol (PEG) as plasticizer. The samples obtained were characterized by Fourier Transformed Infrared Spectroscopy (FTIR), X-Ray Diffraction, Thermal Analysis (TGA/DTG/DSC) and by mechanical characterization with the aim of investigate the interactions between chitosan and clay. From the analysis of the XRD spectrum, it was observed that the so produced nanocomposites presented na intercalated structure. The results obtained from TGA / DTG / DSC techniques showed that a reduction in the thermal stability and glass transition of the produced material occurred. The nanocomposites obtained from biopolymer/sodium bentonite (QuiBen) with PEG 5 % showed better mechanical properties in comparison to the pristine biopolymers due to the enhancement of their ntercalation into the silicate galleries. / Neste trabalho foram preparados nanocompósitos baseados em quitosana ou carborximetilcelulose e diferentes tipos de argilas usando polietileno glicol (PEG) como plastificante. As amostras obtidas foram caracterizadas através de espectroscopia de infravermelho (FTIR), difração de raios X (DRX), análises térmicas (TGA/DTG/DSC), e através de técnicas de análise de tensão e deformação com o objetivo de avaliar as interações entre os biopolímeros, o plastificante e as argilas. Através da analise dos espectros de DRX, foi observado que os nanocompósitos produzidos apresentaram estrutura intercalada. Os resultados obtidos através das técnicas de TGA/DTG/DSC mostraram que houve uma diminuição na estabilidade térmica e na temperatura de transição vítrea do material produzido. Os nanocompósitos obtidos a partir dos biopolímeros/Bentonita sódica com 5 % de PEG apresentaram melhores propriedades mecânicas do que os biopolímeros puros, devido a intercalação dos mesmos nas lamelas da argila.

Este trabalho aborda o estudo de modelos constitutivos a fim de analisar o comportamento mecânico de um biopolímero derivado de óleo de mamona (Ricinus communis). A importância deste trabalho se dá ao fato de: servir como roteiro de ensaio de materiais poliméricos; completar caracterização do material estudado além do regime viscoelástico, bem como aplicação da análise do comportamento do material além do regime viscoelástico linear. A aplicação dos modelos avaliados via Método dos Elementos Finitos (MEF) exigiu o levantamento das propriedades e do comportamento mecânico do biopolímero, bem como a preparação dos ensaios, com as devidas especificações sugeridas pela ASTM (American Society for Testing and Materials) para cada tipo de ensaio. Assim, foram obtidos os corposde- prova (CDP) para a realização dos ensaios mecânicos quase-estáticos de tração, compressão e flexão (monotônicos e cíclicos), bem como, ensaio para análise dinâmicomecânica (Dynamic Mechanical Analysis - DMA) para a obtenção de propriedades associada às parcelas viscosas do material. Com estes foram obtidos parâmetros inerentes a cada tipo de ensaio, permeados com uma cuidadosa revisão bibliográfica sobre as implicações para a realização de cada ensaio. Uma outra etapa foi a verificação de modelos de plastificação baseados na teoria de von Mises, Drucker-Prager e de viscoelasticidade implementados no programa comercial ABAQUS®. Sendo que o objetivo maior foi a comparação dos resultados experimentais obtidos com os modelos em MEF. Os resultados para tração monotônica obtiveram erro de 0,61%, os resultados de compressão monotônica apresentaram erro de 1,5% e as simulações para os carregamentos cíclicos de tração e compressão não foram tão bons quanto os anteriores. Por isso se justifica a criação do modelo fenomenológico utilizando os parâmetros do material que foram identificados. / This work develops the study involving constitutive models with the objective to analyze the mechanical behavior of a biopolymer obtained from the Castor Oil Polyurethane (Ricinus communis). The importance of this work is due to the fact that: serves as a test script for the use of polymer materials; complete the characterization of the material studied beyond the viscoelastic realm, as well as the application of the analysis of the behavior of the material beyond the linear viscoelastic domain. The application of the evaluated models by way of the Finite Element Method (FEM), required the gathering of the properties and mechanical behavior of the biopolymer, as well as the preparation of the tests, with the required specifications suggested by the ASTM (American Society for Testing and Materials) for each type of test. Therefore, there were obtained the body-tests in order to run the mechanical quasi-static traction tests, tensile, compression and flexion (monotonic and cyclical) dynamicmechanical analysis (DMA) in order to obtain the properties associated with the viscous parts of the material. With these the inherent parameters were obtained for each specimen, permeated with a careful biographical revision of the implications for the experimenting of each specimen. Another phase was the verification of laminated models based on the theory by Von Mises, Drucker-Prager and of the viscoelasticity implemented in the commercial program ABAQUS®. The main objective being the comparison of the experimental results obtained with the models in FEM. The results for the monotonic traction obtained a error of 0,61%, the results of monotonic comparison presented an error of 1,5% and the simulations for the cyclical charges of traction and compression were not as good as the former. Therefore it justifies the creation of the phenomenological model utilizing the parameters of the materials that were identified.

L'objectif de ce projet repose sur le développement d’un dispositif médical intra-utérin résorbable prévenant l’apparition ou la récidive d'adhérences intra-utérines (synéchies). La mise en place de ce dispositif suite à chaque agréssion endométriale représenterait un bénéfice symptômatique et économique dans la prise en charge de l’infertilité par trouble de l'implantation. Ce dispositif repose sur l'association originale de polymères résorbables mis sous forme de films stérilisés et maléables, répondants aux contraintes spécifiques de la voie endo-utérine. Des tests sur culture cellulaire ont permis de valider son potentiel anti-adhérentiel en retrouvant des résultats comparables à ceux d’agents barrières disponibles sur le marché. La dégradation in vitro et in vivo du polymère a été étudiée afin de valider un délai d’efficacité suffisant. Des travaux expérimentaux ont été conduits après accord des comités d’éthiques. D’une part, des tests de reproduction animale ont été menés afin de démontrer l’innocuité du dispositif et son efficacité sur la fertilité. D’autre part, des travaux ont permis de valider sur le plan histologique l’effet préventif sur les synéchies. Un travail de modélisation a permis d’adapter la forme du film pour la voie utérine. Les tests de déploiement ont été conduits sur des utérus de cadavre et sur des pièces d’hystérectomie. Après mise en place aisée par les voies naturelles, le dispositif se déploie et gonfle afin de recouvrir la cavité utérine. Des travaux précliniques pourraient être envisagés avant un développement industriel afin d’améliorer les outils disponibles pour la prévention des synéchies intra-utérines. / The objective of this study was to develop an resorbable intra-uterine medical device preventing the appearance or the recurrence of intra-uterine adhesions (synechiaes). The insertion of the device following each endometrial injury would represent a symptomatic and economic benefit in the treatment of infertility by implantation failure. This device is base on the unique combination of resorbables polymers formed into sterilized and malleable films, respondents to the specific constraints of the endo-uterine way. Cellular assays allowed validating the anti-adhesive effect with results comparable to those of currents agents available for the clinical practice. The in vitro and in vivo degradation of the polymer was study to validate a sufficient period of efficiency. The use of experimental models allowed an evaluation of the polymer. On one hand, tests of animal reproduction were lead to demonstrate the harmlessness of the device and the efficiency on the fertility. On the other hand, work has validated histologically preventive effect on synechiaes. Finally, a modeling work allowed adapting the form and dimension of the film for the human uterine way. Tests of deployment carried out on fresh uterus from hysterectomy. After the insertion by the cervical way, the device unfolds and inflates to cover the uterine cavity. Preclinical studies could be done before an industrial development in order to improve the available tools for the clinical prevention of the intra-uterine synechiaes.

La nisine est un peptide antimicrobien produit par la souche de Lactococus lactis subsp. lactis, autorisée pour des applications alimentaires par le comité d'experts sur les additifs alimentaires et les aliments de l'Organisation des Nations Unies pour l’Alimentation et l'Agriculture et de l’Organisation Mondiale de la Santé (FAO / OMS). La nisine peut être appliquée, par exemple, pour la conservation des aliments, la biopréservation, le contrôle de la flore de fermentation, et potentiellement comme agent antimicrobien clinique. Le piégeage de bactéries capables de produire de la nisine dans des billes d'alginate de calcium est ainsi une voie prometteuse pour immobiliser des cellules actives et prolonger la durée de conservation des aliments. Le travail de thèse visait à concevoir des emballages actifs en biopolymère renfermant des bactéries lactiques bioprotectrices (LAB) pour contrôler la croissance de micro-organismes indésirables dans les aliments, en particulier L. monocytogenes. La stabilité mécanique et chimique des billes d'alginate a d’abord été améliorée, et l'efficacité d'encapsulation a été accrue. Des capsules « alginate / pectine (A / P) » ont été préparés comme premières microsphères, par une technique d'extrusion. La production de nisine par Lactococcus lactis subsp. lactis encapsulé dans différents états physiologiques (phase exponentielle, phase stationnaire) a été étudiée. Les résultats ont montré que les billes composites (A/P) avaient de meilleurs propriétés que celles formulées avec de l'alginate ou de la pectine pure. L’association de l'alginate et de la pectine induit un effet synergique qui a amélioré les propriétés mécaniques des microbilles. La deuxième partie du travail a concerné la mise au point de microcapsules à cœur liquides avec une membrane d'hydrogel d'alginate et d'un noyau de gomme de xanthane. Les résultats ont montré que ces microcapsules contenant L. lactis encapsulé durant la phase exponentielle dans une matrice d’alginate et un noyau nutritif de gomme xanthane ont donné les meilleurs résultats et présentent une activité anti-listeria intéressante. Ces microbilles ont enfin été appliquées pour la conservation des aliments et en particulier dans des emballages alimentaires actifs. Des films (HPMC, amidon) ont été élaborés en piégeant des perles d'«alginate actifs/gomme xanthane» enrichies en L. lactis dans des films d’emballage et appliqués pour la conservation des aliments / Nisin is an antimicrobial peptide produced by strains of Lactococus lactis subsp. lactis, recognized as safe for food applications by the Joint Food and Agriculture Organization/World Health Organization (FAO/WHO). Nisin could be applied, for shelf-life extension, biopreservation, control of fermentation flora, and potentially as clinical antimicrobials. Entrapment of bacteria able to produce nisin in calcium alginate beads is promising way for cells immobilization in active films to extend food shelf-life. The present PhD work aimed to design biopolymeric active packaging entrapping bioprotective lactic acid bacteria (LAB) and control undesirable microorganisms growth in foods, particularly L. monocytogenes. First, the mechanical and chemical stability of the alginate beads were improved, and consequently the effectiveness of encapsulation was increased. Alginate/pectin (A/P) biopolymers were prepared, as first microspheres design, by extrusion technique to encapsulate nisin-producing Lactococcus lactis subsp. lactis in different physiological state (exponential phase, stationary phase). Results showed that A/P composite beads were more efficient to increase beads properties than those formulated with pure alginate or pectin. Association of alginate and pectin induced a synergistic effect which improved microbeads mechanical properties. As a second microspheres design, aqueous-core microcapsules were prepared with an alginate hydrogel membrane and a xanthan gum core. Results showed that microcapsules with L.lactis in exponential state encapsulated in alginate membrane and aqueous-core based on xanthan gum with nutrients gave the best results and exhibit interesting antilisterial activity. These microparticles were applied in food preservation and particularly in active food packaging. A novel bioactive films (HPMC, starch) was developed and tested, entrapping active beads of alginate/xanthan gum core-shell microcapsules and alginate/pectin hydrogel enriched with L.lactis

This thesis deals with the study of polyhydroxyalkanoate biosynthesis as a microbial product with the potential to replace current conventional plastics made from petroleum. The dissertation thesis is elaborated in the form of a discussed set of already published publications, which are then part of the thesis in the form of appendices. The work builds on relatively extensive knowledge in the field of polyhydroxyalkanoate production and brings new facts and possible strategies. Various possibilities of analysis of polyhydroxyalkanoates using modern methods were tested in this work, which brings especially speed, which can be crucial in real-time evaluation of production biotechnological process. Raman spectroscopy has proven to be a very promising technique for rapid quantification of PHA. Furthermore, the work deals with valorisation of waste of food and agricultural origin. Emphasis is placed on methods of detoxification of lignocellulose hydrolysates. In this context, adsorption of inhibitors to lignin was first used as an alternative to other detoxification techniques. Due to detoxification, selected production strains Burkholederia cepacia and B. sacchari were able to utilize softwood hydrolyzate for PHA production. In the next part of the work was also tested the possibility of using chicken feathers as a complex source of nitrogen. Evolutionary engineering was also used as a possible strategy to eliminate the inhibitory effect of levulic acid as a microbial inhibitor that results from the hydrolysis of lignocellulosic materials. Adaptation experiments were used to develop strains exhibiting higher resistance to levulic acid and the ability to accumulate a higher 3HV copolymer from the original wild-type C. necator strain. Another promising approach tested in the work was the use of extremophilic microbial strain, which leads to a reduction in the cost of biotechnological production. Selected Halomonas species have shown high potential as halophilic PHA producers. The final part of the thesis was devoted to the selection of the production strain with regard to the properties of the resulting PHA. The Cupriavidus malaysiensis strain was selected to produce a P(3HB-co-3HV-co-4HB) terpolymer which revealed significant differences in material properties over P3HB.

<div>Drug solubility is a persistent challenge in pharmaceutical product development. The objective of this research is to develop a formulation/processing strategy by means of a biodendrimeric solid dispersion (BDSD) platform, for increasing the solubility and dissolution rate of poorly water-soluble drugs. The BSDS platform combines a novel type of excipient, referred to as DLB, with a new application of the hot melt extrusion (HME) process.</div><div><br></div><div>Four model compounds – phenytoin (PHT), griseofulvin (GSF), ibuprofen (IBU), and loratadine (LOR) – were used to evaluate the solubilization effect of an octenylsuccinate-modified dendrimer-like biopolymer (OS-DLB). Shake-flask solubility measurements show that OS-DLB exerts significant solubilizing effect when present at less than 0.2% in water. The presence of hydrophobic C₈ chains on OS-DLB creates the type of favorable nonpolar microenvironment necessary for producing a parallel liquid phase equilibrium responsible for the increase in the total amount of drug dissolved in aqueous media. The higher the hydrophobicity of the drug, the higher the observed solubilization effect. Isothermal titration calorimetry studies show that drug solubilization by OS-DLB occurs by means of entropy-driven interactions. These studies also show that the intermolecular interaction between IBU and OS-DLB in solution exhibits very small energy change upon mixing but a stronger effect on entropy. In comparison, the intermolecular interaction between the less hydrophobic GSF and OS-DLB have significant effects on both enthalpy and entropy. Consequently, in terms of solubilization enhancement, it was found that the interaction between IBU and OS-DLB is entropy-driven (more favorable), while in the case of GSF, the interacting molecules are arranged to maximize enthalpic interaction.</div><div><br></div><div>Based on the solubility studies, a formulation/processing approach for enhancing the dissolution rate of the model drugs was developed. The biopolymer serving as both carrier and solubilizing agent, was coprocessed with poloxamer, functioning as a processing aid, using hot melt extrusion (HME) as an enabling technology. The result is a non-amorphous solid dispersion, exhibiting high and long-lasting supersaturation upon dissolution. A 3-factor, 3-level Box-Behnken design was implemented to define the optimal design space for the formulation/extrusion process. The results obtained from multivariate data analysis (partial least squares and principal components analysis) and response surface modeling suggest that drug release performance of IBU BDSDs is strongly influenced by the processing variables, while maximum release of GSF from the BDSDs can be attained through selective combination of functional excipients.<br></div>

Les polymères issus de la biomasse génèrent depuis quelques années un engouement certain puisqu’ils apparaissent comme de potentiels substituts aux polymères issus de l’industrie pétrolière. Parmi ces monomères récemment développés, l’acide succinique bio-sourcé a reçu une attention particulière notamment pour des applications dans le domaine des polyesters tels que le poly(butylène succinate). La présente thèse décrit la synthèse de ce polymère par estérification directe de l’acide succinique et du 1,4-butanediol dans un réacteur pilote de 7.5 L. Les principaux paramètres du procédé tels que l’excès de diol, la température de trans-estérification ainsi que la pureté de l’acide succinique ont été étudiés. Une attention particulière a été portée sur le choix du catalyseur (son type, la quantité utilisée …) afin d’observer son influence sur le procédé ainsi que sur la stabilité du polymère final. Puis différentes stratégies de modification du poly(butylene succinate) ont été testées pour améliorer à la fois sa mise en forme par extrusion gonflage et les propriétés des films obtenus. Dans ce but l’introduction d’agents de branchements, de silices nanométriques ainsi que des comonomères rigides a été étudiée. / Polymers issued from biomass present a growing interest, since they seem to be a suitable alternative to conventional petrochemical polymers. Among the newly developed monomers, bio-based succinic acid received a particular attention for its application in the synthesis of aliphatic polyesters such as poly(butylene succinate). The present thesis reports the synthesis of this polymer via the direct esterification of succinic acid and 1,4-butanediol in a 7.5 L pilote scale reactor. Main process parameters such as the diol exces, the trans-esterification temperature as well as the purity of succinic acid have been studied. In addition a special attention was taken to highlight the influence of the catalyst (its type, quantity ...) on the synthesis and on the stability of the resulting polymer. Then several strategies of modification of poly(butylene succinate) have been studied to improve the processability of the polymer via film extrusion blowing and to enhance the properties of polymer films. To that end the introduction of branching agents, silica nanofillers as well as rigid comonomers have been studied.