Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: The need for a cost-effective and environmentally friendly substitute for fossil fuels has resulted in significant attention to the production of bioethanol. Lignocellulose being the most abundant renewable resource on the planet consists of cellulose, hemicelluloses and lignin. It can be exploited as a source of fermentable sugars for the conversion to ethanol which may serve as the ultimate fossil fuel replacement. Hemicelluloses, contributing one third of lignocellulose, consists of xylan and mannan. Mannan consists of glucomannan, galactomannan and galactoglucomannan. A cocktail of enzymes are required for its complete hydrolysis, including β-mannanase, β-mannosidase, α-galactosidase, β-glucosidase and acetyl-mannan esterases. A need has arisen for the development of a recombinant microorganism capable of converting lignocelluloses to bioethanol through an economically feasible process. The yeast Saccharomyces cerevisiae naturally ferments hexose sugars into ethanol and has been used in various industrial applications due to its robustness in industrial processes, its well-developed expression systems, its frequent use as a model organism for heterologous gene expression and its current GRAS (Generally Regarded As Safe) status. This yeast is unable to naturally utilise complex lignocelluloses. Recombinant biotechnology can be implemented to overcome this limiting factor. Due to certain restraints by the yeast S. cerevisiae such as hyperglycosylation and poor secretion capacity, alternative hosts such as Aspergillus niger has also been considered for heterologous protein production. The Aspergillus aculeatus β-mannanase (man1) and Talaromyces emersonii α-galactosidase (Agal) genes were expressed in S. cerevisiae Y294. The cDNA of A. niger β-mannosidase (cAnmndA) and synthetic Cellvibrio mixtus β-mannosidase (CmMan5A) were expressed in A. niger. The sequence coding for the native secretion signal from CmMan5A was removed and replaced with the XYNSEC sequence (yielding XYNSEC-CmMan5A) and expressed in E. coli DH5α. The recombinant Man1, Agal, cAnmndA, CmMan5A and XYNSEC-CmMan5A displayed optimal pH of 5.47, 2.37, 3.4, 3.4 and 5.47, respectively, and optimal temperatures of 70°C for Man1, Agal, cAnmndA and CmMan5A and 50°C for XYNSEC-CmMan5A. Activity levels of Man1, Agal, cAnmndA, CmMan5A and XYNSEC-CmMan5A peaked at 36.08, 256.83, 11.61, 7.58 and 2.14 nkat/ml, respectively. Co-expression of Agal and man1 led to a decrease in enzyme secretion and therefore individual expression of these genes should be considered rather than co-expression. The enzymatic activity of Man1, Agal and CmMan5A resulted in a significant decrease in the viscosity of galactomannan when used synergistically. This study confirmed successful production of galactomannan hydrolysing enzymes by the yeast S. cerevisiae and the fungus A. niger, as well as providing insight into the synergistic effect of these enzymes on the viscosity of galactomannan. / AFRIKAANSE OPSOMMING: Die behoefte vir 'n koste-effektiewe en omgewingsvriendelike plaasvervanger vir fossielbrandstowwe het tot 'n beduidende belangstelling in die produksie van bio-etanol gelei. Lignosellulose synde die volopste hernubare hulpbron op die planeet bestaan uit sellulose, hemiselluloses en lignien. Dit kan as 'n bron van fermenteerbare suikers vir die omskakeling na etanol benut word, wat kan dien vir uiteindelike fossielbrandstofvervanging. Hemiselluloses, wat bydra tot 'n derde van lignosellulose, bestaan uit xilaan en mannaan. Mannaan bestaan uit glukomannaan, galaktomannaan en galaktoglukomannaan. 'n Mengsel van ensieme word vir die volledige hidroliese van mannaan benodig, insluitende β-mannanase, β-mannosidase, α-galaktosidase, β-glukosidase en asetiel-mannaan esterases. 'n Behoefte bestaan vir die ontwikkeling van 'n rekombinante mikroörganisme wat in staat is tot die omskakeling van lignoselluloses na bio-etanol deur middel van 'n ekonomies lewensvatbare proses. Die gis Saccharomyces cerevisiae kan heksoe suikers na etanol omskakel en word gebruik in verskeie industriële toepassings as gevolg van sy robuustheid in industriële prosesse, goed ontwikkelde uitdrukking sisteme, gereelde gebruik as 'n model-organisme vir heteroloë uitdrukking van gene en huidige GRAS (Generally Regarded As Safe) status. Die gis is nie daartoe in staat om komplekse lignosellulose te benut nie. Rekombinante biotegnologie kan egter geïmplementeer word om hierdie beperkende faktor te oorkom. As gevolg van sekere beperkinge van die gis S. cerevisiae soos hiperglikosilering en lae sekresie kapasiteit, is alternatiewe gashere soos Aspergillus niger ook oorweeg vir heteroloë proteïenproduksie. Die Aspergillus aculeatus β-mannanase (man1) en Talaromyces emersonii α-galaktosidase (Agal) gene is in S. cerevisiae Y294 uitgedruk. Die cDNA van A. niger β-mannosidase (cAnmndA) en sintetiese Cellvibrio mixtus β-mannosidase (CmMan5A) is in A. niger uitgedruk. Die DNA volgorde wat kodeer vir die natuurlike sekresiesein van CmMan5A is verwyder en vervang met die XYNSEC volgorde (gegewe XYNSEC-CmMan5A) en uitgedruk in E. coli DH5α. Die rekombinante Man1, Agal, cAnmndA, CmMan5A en XYNSEC-CmMan5A vertoon optimale pH kondisies van 5.47, 2.37, 3.4, 3.4 en 5.47, onderskeidelik, en die optimale temperatuur van 70°C vir Man1, Agal, cAnmndA en CmMan5A en 50°C vir XYNSEC-CmMan5A. Aktiwiteitsvlakke van Man1, Agal, cAnmndA, CmMan5A en XYNSEC-CmMan5A het 'n maksimum bereik op 36.08, 256.83, 11.61, 7.58 en 2.14 nkat/ml, onderskeidelik. Gesamentlike uitdrukking van Agal en man1 het tot 'n afname in ensiemsekresie gelei en dus moet individuele uitdrukking van hierdie gene eerder as gesamentlike-uitdrukking oorweeg word. Die ensiematiese aktiwiteite van Man1, Agal en CmMan5A het tot 'n beduidende afname in die viskositeit van galaktomannaan gelei wanneer dit sinergisties gebruik word. Hierdie studie bevestig suksesvolle produksie van galaktomannaan hidrolitiese ensieme in die gis S. cerevisiae en die fungus A. niger, en verskaf insig in die sinergistiese effek van hierdie ensieme op die viskositeit van galaktomannaan.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / Nos Ãltimos anos, um grande interesse na associaÃÃo de materiais magnÃticos e biolÃgicos tem sido relatado na literatura. A obtenÃÃo de novos compÃsitos constituÃdos de galactomanana (GM), nanopartÃculas magnÃticas (MNPs) de NiZn e glicerol (GL) foram produzidos em diferentes proporÃÃes com finalidade de potencializar as caracterÃsticas individuais de cada material para futuras aplicaÃÃes. Sendo assim, as propriedades estruturais, magnÃticas e dielÃtricas dos nanobiocompÃsitos foram investigadas por DifraÃÃo de Raios-X (DRX), Espectroscopia de AbsorÃÃo na RegiÃo de Infravermelho (FTIR), AnÃlise TÃrmica (TG), Calorimetria ExploratÃria Diferencial (DSC), Microscopia EletrÃnica de Varredura (MEV), Microscopia EletrÃnica de TransmissÃo (TEM), Medidas MagnÃticas e Medidas DielÃtricas. A estrutura de espinÃlio da ferrita de NiZn foi confirmada por DRX e TEM e a amostra GMGL apesar de ser um material amorfo apresentou em seus nanobiocompÃsitos picos caracteristicos da fase de NiZn. As bandas caracterÃsticas para as amostras foram confirmadas por FTIR. Estas por sua vez seguiram um perfil de degradaÃÃo de acordo com as quantidades de NiZn incorporados, confirmados nos termogramas de DSC. A caracterizaÃÃo por MEV foi importante para avaliaÃÃo da morfologia. Os resultados das medidas dielÃtricas apresentaram baixas perdas dielÃtricas e das medidas magnÃticas mostraram comportamento magnÃtico para todos os nanobiocompÃsitos. Portanto, os resultados da caracterizaÃÃo dos nanobiocompÃsitos foram satisfatÃrios para possÃveis aplicaÃÃes como biomaterias, dispositivos eletrÃnicos ou em Ãreas afins. / In recent years, a great interest in the association of magnetic and biological materials has been reported in the literature. New composite consisting of galactomannan (GM), magnetic nanoparticles (NPs) of NiZn and glycerol (GL) were produced in different proportions with the purpose of enhancing the individual characteristics of each material for future applications. Thus, the structural, magnetic and dielectric properties of nanobiocomposites were investigated by Absorption Spectroscopy in the Region of Infrared (FTIR), X-Ray Diffraction (XRD), Thermal Analysis (TG), Differential Scanning Calorimetry (DSC), Electron Microscopy (SEM), Transmission Electron Microscopy (TEM), Magnetic Measurements and Dielectric Measurements. The structure of spinel NiZn ferrite was confirmed by XRD and TEM. Sample GMGL despite being an amorphous material presented in their nanobiocomposites characteristic peaks of phase NiZn. The characteristic bands in the samples were confirmed by FTIR. These in turn followed a degradation profile in accordance with the amounts of NiZn incorporated, which was confirmed in the DSC thermograms. The characterization by SEM was important to assess the morphology. The results of dielectric measurements showed low dielectric loss and magnetic measurements showed magnetic behavior for all nanobiocomposites. Therefore, the results of the characterization of nanobiocomposites were satisfactory for potential applications as biomaterials, electronic devices or related areas.

This work seeks to develop a bionanocompÃsito drawn from galactomannan and crystals nanocelulose, both obtained from Prosopis juliflora. Initially, carob pods were subjected to grinding, milling, heating, filtration and centrifugation in order to yield a viscous solution which, with the addition of ethanol, the precipitate was possible galactomannan. So all fractions, and galactomannan fiber capsules were separated. The gum obtained was lyophilized and characterized, as its composition, as well as the fibrous fractions. Further, the fibers of the capsules underwent the hydrothermal process mesquite, followed by a bleaching with H2O2 and NaOH solutions) and acidic hydrolysis (H2SO4 60% v / v) for extracting nanocelulose. After hydrolysis one estÃvl suspension nanocelulose characterized by TGA, XRD, FTIR and zeta potential was obtained. Then galactomannan films were prepared, starting from a 5% (w / v) polysaccharide solution using glycerol as plasticizer. Movies, nanocelulose were added at concentrations of 3, 5 and 7%, and evaluated the influence of the addition of the nanocrystals in the mechanical, thermal and barrier properties to water vapor. The galactomannan extracted from mesquite presented, respectively, in protein, lipid, ash and moisture content of 5.3%, 2.3%, 4.0% and 5.2% and characteristic thermal behavior, showing exothermic event to 284.7  C, typical of his degradation. Their functional groups identified by FTIR, and determination of its mass and molar ratio mannose / galactose. The fiber showed considerable differences in their levels of cellulose, hemicellulose, lignin, insoluble lignin, extractives, ash and moisture, after treatments. These results were corroborated by analysis of FTIR, XRD and TGA. And through the characterizations of the films, it was proved that there were improvements in the thermal, mechanical and barrier properties, the obtained biocomposites, with the addition of nanocelulose. / Este trabalho busca o desenvolvimento de um bionanocompÃsito elaborado a partir de galactomanana e cristais de nanocelulose, ambos obtidos da algaroba (Prosopis juliflora). Inicialmente, vagens da algaroba foram submetidas à trituraÃÃo, moagem, aquecimento, filtraÃÃo e centrifugaÃÃo obtendo-se uma soluÃÃo viscosa onde, com a adiÃÃo de etanol, foi possÃvel precipitar a galactomanana. EntÃo as fraÃÃes de interesse, galactomanana e fibra das cÃpsulas, foram separadas. A goma obtida foi liofilizada e caracterizada, quanto a sua composiÃÃo, assim como as fraÃÃes fibrosas. Na sequÃncia, as fibras das cÃpsulas da algaroba sofreram processo hidrotÃrmico, seguido de dois branqueamentos com soluÃÃes de NaOH e H2O2, e hidrÃlise Ãcida (H2SO4 60% v/v) para extraÃÃo de nanocelulose. ApÃs a hidrÃlise, foi obtida uma suspensÃo estÃvel de nanocelulose, caracterizada por TGA, DRX, potencial zeta e FTIR. EntÃo foram preparados filmes de galactomanana, a partir de uma soluÃÃo 5% (m/v) do polissacarÃdeo, utilizando glicerol como plastificante. Aos filmes, foram adicionados nanocelulose, nas concentraÃÃes de 3, 5 e 7%, e avaliado a influÃncia da adiÃÃo dos nanocristais nas propriedades mecÃnicas, tÃrmicas e de barreira a vapor de Ãgua. A galactomanana extraÃda da algaroba apresentou, respectivamente, teores de proteÃna, lipÃdio, cinzas e umidade de 5,3%, 2,3%, 4,0% e 5,2% e comportamento tÃrmico caracterÃstico, apresentando evento exotÃrmico a 284,7ÂC, tÃpico de sua degradaÃÃo. Seus grupos funcionais identificados por FTIR, alÃm da determinaÃÃo de sua massa molar e razÃo manose/galactose. A fibra apresentou considerÃveis diferenÃas nos seus teores de celulose, hemicelulose, lignina solÃvel, lignina insolÃvel, extrativos, cinzas e umidade, apÃs os tratamentos. Esses resultados foram corroborados por anÃlises de FTIR, DRX e TGA. AtravÃs das caracterizaÃÃes dos filmes, ficou comprovado que a adiÃÃo de nanocelulose melhorou propriedades tÃrmicas, mecÃnicas e de barreira, dos biocompÃsitos obtidos.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / Devido Ãs suas diferentes propriedades quÃmicas tais como capacidade de formar soluÃÃes viscosas ou gÃis em meio aquoso o isolamento e caracterizaÃÃo de galactomananas tÃm sido de grande importÃncia Nesse trabalho a galactomanana do endosperma da semente de Bauhinia pentandra foi isolada e caracterizada, apresentando-se homogÃnea por GPC Este polissacarideo foi demonstrado ser uma galactomanana clÃssica formada por uma cadeia linear de manose unidas por ligaÃÃes β(1-4) com substituiÃÃes de galactose em ligaÃÃo α(1-6) com uma proporÃÃo Man:Gal de 2 5.1 e viscosidade intrÃnseca em Ãgua de 10 1 dL/g A galactomanana foi avaliada quanto à capacidade de interagir com lectinas galactose ligante O polissacarÃdeo foi tratado com epicloridrina e o material obtido foi utilizado para a montagem de coluna cromatogrÃfica de afinidade Extratos ricos em lectinas de Artocarpus incisa Artocarpus integrifÃlia e Bauhinia pentandra foram aplicados e fraÃÃes lectinicas purificadas foram obtidas A capacidade da galactomanana de B. pentandra em reter a lectina (LBp) da mesma semente foi comparada com a matriz de galactomanana de Adenantera pavonina Caesalpinea pulcherrima Sophora japonica e com a matriz comercial Sepharose 4B Apesar da galactomanana de B. pentandra ter apresentado a menor capacidade de retenÃÃo frente Ãs demais, ela mostrou-se semelhante à matriz comercial sendo viÃvel a sua utilizaÃÃo / Due the chemicals properties diferences, such as the ability to make viscous solution or aqueous gels, the study of the galactomanans has been too important. In this study, the endospermic galactomannans from seeds of Bauhinia pentandra was isolated and partially characterized. This polysaccharide is a classical galalactomannan constituted by a linear chain of mannose linked by β(1-4) linkages with galactose substituintions linked by α(1-6) linkages, resulting in a Man:Gal ratio of 2.5:1, and water intrinsic viscosity equal to 10.1 dL/g. Galactomannan was evaluated in ability to interact with galactose-binding lectins. The polysaccharide was treated with epichlorohidrine and the material obtained was utilized to make the affinity chromatography matrix. Lectin-rich extracts from Artocarpus incisa, Artocarpus integrifolia and Bauhinia pentandra were applied and lectin fractions were obtained, thus, the affinity matrix showed to be efficient to isolate them. The retention capacity of the galactomannan from B. pentandra was compared with galactomannan matrix from Adenantera pavonina, Caesalpinea pulcherrima, Sophora japonica and commercial matrix of Sepharose 4B in regards to the isolation of the lectin from B. pentandra (LBp). Although the galactomannan matrix had been showed the smallest retention capacity in comparision with the others, it is equivalent to the commercial matrix, enabling your utilization

Diese Arbeit untersucht das Auftreten falschpositiver Mannan- und Galactomannanergebnisse bei hämatologischen Patienten nach allogener Stammzelltransplantation, die parenteral ernährt werden müssen oder eine antibiotische Therapie mit Piperacillin-Tazobactam erhalten. Die Kenntnis von Quellen, die die Spezifität des Platelia™ Candida Ag plus und den Platelia™ Aspergillus EIA beeinflussen, ist ein wichtiger Beitrag zur Diagnostik systemischer Pilzinfektionen. / This study investigates the occurrence of false-positive mannan and galactomannan results in hematological patients undergoing allogeneic hematopoietic cell transplantation with current parenteral nutrition or antibiotic treatment with piperacillin-tazobactam. The knowledge of sources influencing specificity of Platelia™ Candida Ag plus and Platelia™ Aspergillus EIA contributes to the diagnosis of invasive fungal infections.

MESQUITA, Ricardy Leonam Pontes. Síntese de nanopartículas de prata através de ultrassom utilizando galactomanana. 2016. 94 f. Dissertação (Mestrado em química)- Universidade Federal do Ceará, Fortaleza-CE, 2016. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-07-28T17:12:54Z No. of bitstreams: 1 2016_dis_rlpmesquita.pdf: 2439147 bytes, checksum: 18f53a2e3356343fac06a5df49665f6e (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-08-02T14:38:19Z (GMT) No. of bitstreams: 1 2016_dis_rlpmesquita.pdf: 2439147 bytes, checksum: 18f53a2e3356343fac06a5df49665f6e (MD5) / Made available in DSpace on 2016-08-02T14:38:19Z (GMT). No. of bitstreams: 1 2016_dis_rlpmesquita.pdf: 2439147 bytes, checksum: 18f53a2e3356343fac06a5df49665f6e (MD5) Previous issue date: 2016 / Silver nanoparticles (NPAg) have attracted considerable interest in the context of research in recent years due to numerous applications, such as a bactericide, sensors and catalysts. In this paper the use of galactomannan of fava danta (GFD) in obtaining NPAg by ultrasonic method was studied based on the principles of green chemistry without the use of conventional chemical reducing agent. This study aimed to evaluate the ultrasound employment in NPAg synthesis GFD solutions. The NPAg were obtained with the aid of an ultrasonic probe using AgNO3 aqueous solution in concentrations of 10, 40 and 100 mmol / L. The galactomannan in aqueous solution (0,032%, 0,32% and 0,0032% w/ v) was used as stabilizer of NPAg. The synthesis occurred at varying pH (10, 11, 12) and temperature (15, 25 and 35 °C). The characterizations were performed using techniques such as gel permeation chromatography (GPC), nuclear magnetic resonance of hydrogen (NMR1H), spectrophotometry UV-Vis, dynamic light scattering (DLS) and scanning electron microscopy (SEM). UV-Vis analysis for colloids formed during the synthesis at the three concentrations showed the emergence of the plasmon band (around 400 nm) suggesting the formation of NPAg. It was observed faster kinetic profile for concentration systems 10 mmol/ L AgNO3. The pH 11 and 12 provided the best conditions for synthesis of NPAg. The different concentrations of synthetic GFD showed satisfactory results, but the formed colloid concentration 0,0032% w/ v showed no stability. Two of the synthesized colloids were selected FDAg 10/12/25 and FDAg 100/12/25, to submit the study to estimate size during the time of formation of NPAg as were also objects of analysis by electron microscopy scan. DLS was evidenced by the two colloids have decreased the size NPAg during the synthesis time, reaching average sizes near 10 to 4 nm for FDAg 10/12/25 and FDAg 100/12/25 respectively. Monitoring the polysaccharide degradation by GPC analysis was conducted and it was observed the decrease in distribution of the molar masses of the same of 6,65 x 106 to 3,11 x 104 g/ mol during the degradation process. The mannose/ galactose ratio was observed for the GFD, from 1:1,23 to GFD unmodified to 1:2,14 for the GFD degraded in 420 minutes. Thus, this study showed that the ultrasonic path in conjunction with the use of the galactomannan of fava danta in aqueous solution provides a versatile synthetic route for NPAg with good stability. / Nanopartículas de prata (NPAg) têm atraído bastante interesse no âmbito da pesquisa durante os últimos anos devido às inúmeras aplicações, tais como agente bactericida, sensores e catalisadores. Neste trabalho o uso de galactomanana da fava danta (GFD) na obtenção das NPAg por método ultrassônico foi estudado com base nos princípios da química verde sem a utilização de redutores químicos convencionais. Este estudo teve como objetivo, avaliar o emprego do ultrassom na síntese de NPAg em soluções de GFD. As NPAg foram obtidas com auxílio de uma sonda ultrassônica, utilizando solução aquosa de AgNO3 nas concentrações 10, 40 e 100 mmol/L. A galactomanana em solução aquosa (0,032%, 0,32% e 0,0032% m/v) foi utilizada como estabilizante das NPAg. As sínteses ocorreram com variação de pH (10, 11 e 12) e de temperatura (15, 25 e 35 °C). As caracterizações foram realizadas através de técnicas como Cromatografia de permeação em gel (GPC), Ressonância magnética nuclear de hidrogênio (RMN 1H), Espectrofotometria UV-Vis, Espalhamento de luz dinâmico (DLS) e Microscopia eletrônica de varredura (MEV). As análises de UV-Vis para os coloides formados durante as sínteses nas três concentrações mostraram o surgimento da banda de plasmon (em torno de 400 nm) evidenciando a formação das NPAg. Observou-se perfil cinético mais rápido para os sistemas de concentração 10 mmol/L de AgNO3. Os valores de pH 11 e 12 proporcionaram as melhores condições de síntese das NPAg. As diferentes concentrações de GFD apresentaram resultados de síntese satisfatórios, porém o coloide formado em concentração 0,0032% m/v não apresentou estabilidade. Dois dos coloides sintetizados foram selecionados, FDAg 10/12/25 e FDAg 100/12/25, a se submeterem a estudo para estimativa de tamanho durante todo o tempo de formação das NPAg, assim como também foram objetos de análise por Microscopia eletrônica de varredura. Por DLS foi evidenciado que os dois coloides tiveram diminuição de tamanho das NPAg no decorrer do tempo de síntese, atingindo tamanhos médios próximos de 10 e 4 nm para FDAg 10/12/25 e FDAg 100/12/25 respectivamente. Foi realizado o acompanhamento da degradação do polissacarídeo através de análise por GPC, tendo sido observado a diminuição da distribuição das massas molares do mesmo de 6,65 x 106 para 3,11 x 104 g/mol durante o processo de degradação. A razão manose/galactose foi verificada para a GFD, passando de 1:1,23 para a GFD não modificada para 1:2,14 para a GFD degradada em 420 minutos. Sendo assim, este estudo mostrou que a via ultrassônica em conjunto com a utilização da galactomanana da fava danta em solução aquosa proporciona uma rota sintética bastante versátil para as NPAg com boa estabilização.

Universidade Federal do Cearà / Silver nanoparticles (NPAg) have attracted considerable interest in the context of research in recent years due to numerous applications, such as a bactericide, sensors and catalysts. In this paper the use of galactomannan of fava danta (GFD) in obtaining NPAg by ultrasonic method was studied based on the principles of green chemistry without the use of conventional chemical reducing agent. This study aimed to evaluate the ultrasound employment in NPAg synthesis GFD solutions. The NPAg were obtained with the aid of an ultrasonic probe using AgNO3 aqueous solution in concentrations of 10, 40 and 100 mmol / L. The galactomannan in aqueous solution (0,032%, 0,32% and 0,0032% w/ v) was used as stabilizer of NPAg. The synthesis occurred at varying pH (10, 11, 12) and temperature (15, 25 and 35 ÂC). The characterizations were performed using techniques such as gel permeation chromatography (GPC), nuclear magnetic resonance of hydrogen (NMR1H), spectrophotometry UV-Vis, dynamic light scattering (DLS) and scanning electron microscopy (SEM). UV-Vis analysis for colloids formed during the synthesis at the three concentrations showed the emergence of the plasmon band (around 400 nm) suggesting the formation of NPAg. It was observed faster kinetic profile for concentration systems 10 mmol/ L AgNO3. The pH 11 and 12 provided the best conditions for synthesis of NPAg. The different concentrations of synthetic GFD showed satisfactory results, but the formed colloid concentration 0,0032% w/ v showed no stability. Two of the synthesized colloids were selected FDAg 10/12/25 and FDAg 100/12/25, to submit the study to estimate size during the time of formation of NPAg as were also objects of analysis by electron microscopy scan. DLS was evidenced by the two colloids have decreased the size NPAg during the synthesis time, reaching average sizes near 10 to 4 nm for FDAg 10/12/25 and FDAg 100/12/25 respectively. Monitoring the polysaccharide degradation by GPC analysis was conducted and it was observed the decrease in distribution of the molar masses of the same of 6,65 x 106 to 3,11 x 104 g/ mol during the degradation process. The mannose/ galactose ratio was observed for the GFD, from 1:1,23 to GFD unmodified to 1:2,14 for the GFD degraded in 420 minutes. Thus, this study showed that the ultrasonic path in conjunction with the use of the galactomannan of fava danta in aqueous solution provides a versatile synthetic route for NPAg with good stability. / NanopartÃculas de prata (NPAg) tÃm atraÃdo bastante interesse no Ãmbito da pesquisa durante os Ãltimos anos devido Ãs inÃmeras aplicaÃÃes, tais como agente bactericida, sensores e catalisadores. Neste trabalho o uso de galactomanana da fava danta (GFD) na obtenÃÃo das NPAg por mÃtodo ultrassÃnico foi estudado com base nos princÃpios da quÃmica verde sem a utilizaÃÃo de redutores quÃmicos convencionais. Este estudo teve como objetivo, avaliar o emprego do ultrassom na sÃntese de NPAg em soluÃÃes de GFD. As NPAg foram obtidas com auxÃlio de uma sonda ultrassÃnica, utilizando soluÃÃo aquosa de AgNO3 nas concentraÃÃes 10, 40 e 100 mmol/L. A galactomanana em soluÃÃo aquosa (0,032%, 0,32% e 0,0032% m/v) foi utilizada como estabilizante das NPAg. As sÃnteses ocorreram com variaÃÃo de pH (10, 11 e 12) e de temperatura (15, 25 e 35 ÂC). As caracterizaÃÃes foram realizadas atravÃs de tÃcnicas como Cromatografia de permeaÃÃo em gel (GPC), RessonÃncia magnÃtica nuclear de hidrogÃnio (RMN 1H), Espectrofotometria UV-Vis, Espalhamento de luz dinÃmico (DLS) e Microscopia eletrÃnica de varredura (MEV). As anÃlises de UV-Vis para os coloides formados durante as sÃnteses nas trÃs concentraÃÃes mostraram o surgimento da banda de plasmon (em torno de 400 nm) evidenciando a formaÃÃo das NPAg. Observou-se perfil cinÃtico mais rÃpido para os sistemas de concentraÃÃo 10 mmol/L de AgNO3. Os valores de pH 11 e 12 proporcionaram as melhores condiÃÃes de sÃntese das NPAg. As diferentes concentraÃÃes de GFD apresentaram resultados de sÃntese satisfatÃrios, porÃm o coloide formado em concentraÃÃo 0,0032% m/v nÃo apresentou estabilidade. Dois dos coloides sintetizados foram selecionados, FDAg 10/12/25 e FDAg 100/12/25, a se submeterem a estudo para estimativa de tamanho durante todo o tempo de formaÃÃo das NPAg, assim como tambÃm foram objetos de anÃlise por Microscopia eletrÃnica de varredura. Por DLS foi evidenciado que os dois coloides tiveram diminuiÃÃo de tamanho das NPAg no decorrer do tempo de sÃntese, atingindo tamanhos mÃdios prÃximos de 10 e 4 nm para FDAg 10/12/25 e FDAg 100/12/25 respectivamente. Foi realizado o acompanhamento da degradaÃÃo do polissacarÃdeo atravÃs de anÃlise por GPC, tendo sido observado a diminuiÃÃo da distribuiÃÃo das massas molares do mesmo de 6,65 x 106 para 3,11 x 104 g/mol durante o processo de degradaÃÃo. A razÃo manose/galactose foi verificada para a GFD, passando de 1:1,23 para a GFD nÃo modificada para 1:2,14 para a GFD degradada em 420 minutos. Sendo assim, este estudo mostrou que a via ultrassÃnica em conjunto com a utilizaÃÃo da galactomanana da fava danta em soluÃÃo aquosa proporciona uma rota sintÃtica bastante versÃtil para as NPAg com boa estabilizaÃÃo.

Orientador: Marcos Silveira Buckeridge / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-12T18:38:24Z (GMT). No. of bitstreams: 1 Lisboa_CesarGustavoSerafim_D.pdf: 1945488 bytes, checksum: 831197d663c65044a02bdfd7ba13598f (MD5) Previous issue date: 2008 / Resumo: Aplicações biotecnológicas de polissacarídeos de plantas: uma estratégia sustentável para a produção em demanda. Atualmente, a busca por novas fontes de materiais pode levar ao depósito de patentes. Isto é importante para que novos produtos, que apresentem características inovadoras, potencial de redução de custos, rotas de produção ambientalmente corretas, partição de benefícios e, finalmente, possibilitem a exploração de novos mercados de maneira sustentável. Nesse contexto, o presente trabalho teve como objetivo avaliar o potencial de uso de polissacarídeos de plantas de diferentes fontes nos setores de alimentos, farmacosméticos e papel. Os resultados são apresentados na forma de um artigo e 4 patentes. Os xiloglucanos de sementes de espécies nativas de plantas (Hymenaea courbaril) e de plântulas de eucalipto foram aplicados no processo de confecção do papel, demonstrando que suas propriedades mecânicas podem ser melhoradas. Resultados similares foram obtidos com galactomananos de sementes de Sesbania virgata e Dimorphandra mollis. O xiloglucano de H. courbaril foi estudado mais profundamente e demonstrou-se que pode ser também aplicado em cosméticos, pois não é citotóxico, é capaz de espessar formulações e interfere na produção de colágeno pela pele. As propriedades reológicas do galactomanano de D. mollis foram estudadas em comparação a outros galactomananos já utilizados na indústria. Demonstrou-se que ele é apropriado para várias aplicações em alimentos. Como um todo, nossos resultados indicam que tanto os polissacarídeos de sementes de espécies nativas como o de eucalipto podem ser utilizados em várias aplicações de tal forma que possam fazer parte de programas de uso sustentável da biodiversidade, levando a benefícios tecnológicos e também sociais. / Abstract: Biotechnological applications of plant polysaccharides: a sutainable strategy for production and demand. Nowdays, the search for new sources of materials can lead to new patent deposits. This is important because new products that display innovative features, cost reduction potential, environmentally clean production routes, partition of benefits and finally, may turn possible to explore differential market shares. These may be excellent socio-environmental and market shares for biotechnology companies. In this context, the present work aimed at evaluating the potential of use of plant polysaccharides from different sources in the food, farmacosmetic and paper industrial sectors. In the present work, we present studies that focus on the use of seed polysaccharides on applications on paper, cosmetics and food industries. The results are presented in the form of one article and 4 patents. The xyloglucan from seeds of native tropical tree species (Hymenaea courbaril - jatobá, Copaifera langsdorffii - copaiba) and from seedlings of Eucalyptus grandis (eucalypt) were applied to paper making and we demonstrated that several properties of paper can be improved by using these xyloglucans during preparation. Similar results were obtained with galactomann ans from Sesbania virgata and Dimorphandra mollis, two fast-growing legume trees. The polymers increased paper resistance to mechanical stress so that they can be applied for applications where more resistant papers (such as rapping paper) are used. The xyloglucan from H. courbaril was studied more deeply, demonstrating that it can be applied in cosmetics as it is not cytotoxic and can be used to improve skin biochemistry such as collagen production. Galactomannan from D. mollis was studied from the rheological point of view and its properties were compared with other commercially used galactomannans. Our results showed that D. mollis galactomannan is appropriate to several applications in food industries. As a whole, our results indicate that both, native seeds and seeds of eucalyptus, might be used in several industrial applications so that these might be used as part of programs of sustainable use of biodiversity, leading to social and technological benefits to society. / Doutorado / Biologia Celular / Doutor em Biologia Celular e Estrutural

Astragalus gombo Bunge (Fabaceae) est une plante bien représentée dans le Sahara Septentrional Est Algérien (région de Ouargla, Algérie) et couramment utilisée comme fourrage pour les animaux ou en médecine traditionnelle. Malgré de nombreuses publications sur la richesse en polysaccharides d’autres espèces appartenant au genre Astragalus et la description de leurs propriétés biologiques putatives ou avérées, aucune étude ne s’est intéressée à l’espèce gombo. L’objectif de ce travail de thèse a donc été de réaliser des investigations sur plusieurs parties de cette plante en vue d’extraire, identifier et caractériser son contenu en polysaccharides. Dans un second temps les propriétés biologiques et rhéologiques de ces polymères ont été étudiées afin d’identifier d’éventuelles voies de valorisation. Les résultats obtenus ont permis de détecter la présence de composés pectiques et hémicellulosiques dans la tige d’Astragalus gombo et celle d’un galactomannane dans les graines. Le galactomannane est de haute masse moléculaire (1,1x106 Da) et est constitué d’une chaine principale de β-(1→4)-D-mannane ramifiée en α-(1→6) par des résidus Dgalactopyranoses avec un rapport M/G de 1,7. La caractérisation de ses propriétés rhéologiques a révélé un comportement typique rhéofluidifiant et des propriétés viscoélastiques. L’étude des activités biologiques associées à ce biopolymère a révélé son potentiel comme prébiotique et antioxydant. / Astragalus gombo Bunge (Fabaceae) is a terrestrial plant occuring in the East Septentrional Sahara (Ouargla, Algeria). It is commonly used as fodder or in traditional medicine by local populations. Despite numerous publications focusing on polysaccharidic contents of Astragalus species and the designation of their putative or proved biological activities, no study has examined those of A. gombo. The objective of this thesis was firstly to investigate several organs of this plant for their polysaccharide contents. In a second step, the biological and rheological properties of these biopolymers have been studied to identify ways of adding value. Results led to the identification of pectic compounds and hemicelluloses in the rods of Astragalus gombo whereas a galactomannan was detected in its seeds. This galactomannan was a high molecular weight macromolecule composed of a β-(1→4)-D-mannan skeleton ramified by residues of D-galactopyranoses. The M/G ratio was of 1.7. The characterization of its rheological behavior was typic of that of a rheofluidifiant fluid with viscoelastic properties. The study of its biological properties showed its potential as prebiotic and antioxidant agent.

Invasive fungal infections (IFIs) are associated with considerable morbidity and mortality among high-risk individuals. Outcomes for IFI historically have been suboptimal and associated with a high mortality rate, hence global prophylaxis strategies have been applied to at-risk populations. Among certain populations, fluconazole prophylaxis has reduced systemic and superficial infections caused by Candida species. Newer azoles are currently being evaluated as prophylaxis and have the potential to provide protection against mould pathogens that are more troublesome to treat once they occur. Global prophylaxis strategies have the shortcoming of subjecting patients to therapy that ultimately will not need it. Targeted prophylaxis has the advantage of treating only patients at highest risk using some parameter of greater host susceptibility. Prophylaxis strategies are most suitable in patients at the highest risk for IFI. For patient groups whose risk is somewhat lower or when suspicion of IFI occurs in patients receiving prophylaxis, empirical antifungal therapy is often employed following a predefined period of fever. Again this approach subjects many non-infected patients to unnecessary and toxic therapy. A more refined approach such as presumptive or pre-emptive therapy whereby treatment is only initiated upon positive identification of a surrogate marker of infection in combination with clinical and radiological signs will subject fewer patients to toxic and expensive treatments.