Engineering of fatty acid production and secretion in Saccharomyces cerevisiae

Tese de Doutoramento em Ciências - Especialidade em Biologia

Mechanism of extracellular silver nanoparticles synthesis by Stereum hirsutum and Fusarium oxysporum

The increasing interest for greener and biological methods of synthesis has led to the development of non-toxic and comparatively more bioactive nanoparticles. Unlike physical and chemical methods of nanoparticle synthesis, microbial synthesis in general and mycosynthesis in particular is cost-effective and environment-friendly. However, different aspects, such as the rate of synthesis, monodispersity and downstream processing, need to be improved. Many fungal-based mechanisms have been proposed for the formation of silver nanoparticles (AgNPs), mainly those involving the presence of nitrate reductase, which has been detected in filtered fungus cell used for AgNPs production. There is a general acceptance that nitrate reductase is the main responsible for the reduction of Ag ions for the formation of AgNPs. However, this generally accepted mechanism for fungal AgNPs production is not totally understood. In order to elucidate the molecules participating in the mechanistic formation of metal nanoparticles, the current study is focused on the enzymes and other organic compounds involved in the biosynthesis of AgNPs. The use of each free fungal mycelium of both Stereum hirsutum and Fusarium oxysporum will be assessed. In order to identify defective mutants on the nitrate reductase structural gene niaD, fungal cultures of S.hirsutum and F.oxysporum will be selected by chlorate resistance. In addition, in order to verify if each compound identified as key-molecule influenced on the production of nanoparticles, an in vitro assay using different nitrogen sources will be developed. Lately, fungal extracellular enzymes will be measured and an in vitro assay will be done. Finally, The nanoparticle formation and its characterization will be evaluated by UV-visible spectroscopy, electron microscopy (TEM), X-ray diffraction analysis (XRD), Fourier transforms infrared spectroscopy (FTIR), and LC-MS/MS.

Synthesis, physical and magnetic properties of BaFe12O19/P(VDF-TrFE) multifunctional composites

Composite films with filler microparticles of Barium ferrite dispersed within P(VDF-TrFE) as polymeric matrix have been prepared by solvent evaporation. The lowest BaFO content of 1% wt acts as a small defect within the polymeric matrix, reducing the values of the dielectric and mechanical properties of the pure P(VDF-TrFE). For filler contents up to a 20%, the BaFO filler reinforces the matrix and measured properties increase their values. This trend is not followed by the electrical conductivity. We extended the study to fibers composed by BaFe12O19 microparticles in a PVDF matrix. Due to the big size of BaFO particles (1 micron in diameter), proper fabrication of the fiber shaped composites has not been achieved. We found that true BaFO content are always lower than nominal ones. Results are discussed in terms of the influence of size and morphology of the BaFO particles on the initial properties of the polymeric matrix.

One-step in situ synthesis of polyamide microcapsules with inorganic payload and their transformation into responsive thermoplastic composite materials

Well-dispersed loads of finely powdered metals, metal oxides, several carbon allotropes or nanoclays are incorporated into highly porous polyamide 6 microcapsules in controllable amounts via an original one-step in situ fabrication technique. It is based on activated anionic polymerization (AAP) of ε-caprolactam in a hydrocarbon solvent performed in the presence of the respective micro- or nanosized loads. The forming microcapsules with typical diameters of 25-50 µm entrap up to 40 wt% of load. Their melt processing produces hybrid thermoplastic composites. Mechanical, electric conductivity and magnetic response measurements show that transforming of in situ loaded microcapsules into composites by melt processing (MP) is a facile and rapid method to fabricate materials with high mechanical resistance and electro-magnetic characteristics sufficient for many industrial applications. This novel concept requires low polymerization temperatures, no functionalization or compatibilization of the loads and it is easy to scale up at industrial production levels.

Aproveitamento do Óleo das Amêndoas de Tucumã do Amazonas na Produção de Biodiesel

A falta de disponibilidade de energia elétrica é um dos principais motivos pelo baixo Índice de Desenvolvimento Humano das comunidades isoladas localizadas na Amazônia. O biodiesel produzido a partir de óleos vegetais extraídos de espécies oleaginosas nativas de forma sustentada é uma das melhores alternativas energéticas para a região. O tucumã do amazonas, Astrocaryum aculeatum, é uma espécie de palmeira que produz um fruto muito apreciado na região, a partir do qual se obtém uma amêndoa com alto teor de óleo. Nesse estudo, foi avaliada a produção de biodiesel etílico, a partir de diferentes lotes de óleos de tucumã do amazonas, com índices de acidez baixos e elevados, pela transesterificação por catálise básica e ácida homogêneas, respectivamente. Na catálise ácida, foram testados HCl e H2SO4 como catalisadores nas concentrações de 0,0625 a 1,000 M, empregando etanol hidratado na proporção molar de 1:6 e a reação conduzida a 90 ºC por 24 h. Na catálise básica, foram testados NaOH e KOH, nas proporções de 0,5 a 2,0 %, empregando etanol anidro na proporção molar de 1:12 e a reação conduzida a 80 ºC por 2 h. O biodiesel obtido em cada experimento foi analisado por métodos físicos (massa específica) e cromatográficos (CLAE em fase reversa). Análises cromatográficas indicaram que as melhores conversões foram alcançadas por amostras de biodiesel com massas específicas inferiores a 0,87 g.cm-1. As amostras de biodiesel obtidas com melhor qualidade foram obtidas utilizando-se os catalisadores ácidos a 1,0 M com rendimentos superiores a 90%. No caso da catálise básica, obteve-se biodiesel de boa qualidade empregando-se o catalisador NaOH a 2,0%, porém com rendimento inferior a 60 %. Contudo, em ambos os casos, foi possível identificar um excelente potencial de produção de biocombustível, a partir do óleo das amêndoas de tucumã.

Caracterização físico-química das misturas binárias de biodiesel e diesel comercializados no Amazonas

O uso de energias renováveis aliadas à preservação do meio ambiente tem despertado interesse cada vez maior em nível nacional e internacional principalmente a utilização do biodiesel em substituição ao diesel de petróleo. Sua utilização traz uma série de vantagens sociais, econômicas e ambientais. Nos últimos anos, houve no Brasil um aumento nas pesquisas sobre o biodiesel. A lei brasileira nº 11.097 de 2005 dispõe sobre a introdução do biodiesel na matriz energética brasileira, misturado ao óleo diesel em um percentual mínimo obrigatório de 2%, com aumento na adição para 5% até 2012. Contudo, essa adição só é possível se o biodiesel estiver dentro dos padrões de qualidade exigidos. Diante disto, neste trabalho foram preparadas misturas de biodiesel adicionado ao diesel interior, nas proporções 2, 5, 10 e 20% (v/v), sendo posteriormente caracterizados a partir da determinação dos parâmetros físico-químicos citados na Resolução da Agência Nacional do Petróleo, Gás Natural e Biocombustíveis (ANP), visando avaliar se as misturas estão em conformidade com a norma. De acordo com os resultados concluiu-se que, tanto o biodiesel quanto suas misturas binárias (B2 a B20) encontram-se dentro dos padrões de qualidade estabelecido pela ANP.

Synthesis and anticancer activity of N3-substituted-6,8-diaminopurines

[Excerpt] Cancer is the second most common cause of death in developed countries appearing just after cardiovascular diseases. The treatment of cancer remains a major medical challenge and the development of new anticancer drugs is an emerging topic for the scientific community. During the past three decades several chemical classes of anticancer drugs have been identified. In particular, 2,6-diamino purines proved to be important candidates as new anti-cancer agents.

A new and efficient synthesis of N3-cycloalkylamino-6,8-diaminopurines

[Excerpt] The purine core is a privileged scaffold in medicinal chemistry and the biological relevance of purine derivatives makes them attractive targets in the preparation of combinatorial libraries.1,2 In particular, there is a great interest in the synthesis of 8-substituted purines due to their important potential as antiviral and anticancer agents.3 Reports on 8-aminopurines are limited and general methods to obtain these purine derivatives are still needed.4 Cyclic amines and hydrazines are key structural motifs in various bioactive agents.5 Here we report a novel, efficient and inexpensive method for the synthesis of 6,8-diaminopurines 4 incorporating cycloalkylamino substituents at N3position of the purine ring. (...)

A new and efficient synthesis of benzoimidazopurine derivatives

[Excerpt] Purine nucleobases are essential biomolecules in living organisms. Playing several key roles in the cell, they have been a significant inspiration for drug design.1 Benzimidazole nucleus is an important pharmacophore in the development of molecules with pharmaceutical or biological interest. Benzimidazoles have been reported to display significant pharmacological activities such as antiulcer, antifungal, antiparkinson, anticancer and antibiotic.2 Fused structures incorporating these two scaffolds might be important for medicinal chemistry and, to the best of our knowledge, there are no reports of these systems in the literature. In particular, benzo[4,5]imidazo[2,1]purines seem to be novel and must be important target molecules in the heterocyclic synthesis. (...)

The synthesis of new 6-hydrazinopurines as potential anticancer agents

[Excerpt] Purines, such as adenine, are one of the most important naturally occurring nitrogen heterocycles and they are frequently used as bioactive agents.[1,2] The increasing number of synthetic purines reveals the great potential of these compounds as enzyme inhibitors. Protein Kinases have an important regulatory role in cell proliferation, differentiation and signalling processes. Abnormal signal transduction is responsible for devastating diseases such as cancer. All of the protein kinases identified have in common the cofactor ATP indicating that the adenine nucleus is a very important scaffold for discovery of new anti-cancer agents.[3,4] Previous work identified a modest anticancer activity in a family of 6-arylaminopurines. In the view of these results, it seemed reasonable to assume that some interesting anticancer agents might result by replacement of the phenyl group by a secondary amino group linked to the N-6 atom of the adenine moiety. (...)

The synthesis of novel 5-aminoimidazoles derivatives with anticancer activity

[Excerpt] The imidazole nucleus is present in a significant number of biomolecules and the inclusion of this moiety in organic scaffolds is considered an important synthetic strategy in drug discovery.[1] 5-Aminoimidazoles are interesting building blocks in medicinal chemistry since they are key components in many bioactive molecules and their derivatives showed a wide pharmacological potential as anticancer drugs.[1] The hydrazones constitute an important class of biological active drug molecules due to their wide range of pharmacological properties that include antitumoral activities.[2] Amidrazone derivatives could be considered very promising in the perspective of new drug discovery, because they are very effective as building blocks to obtain various heterocycles.[2,3] The α-hydrazononitriles are a special case of compounds belonging to the family of hydrazones that is less common in the literature, but has a great interest due to their pharmacological applications.[4] (...)

Peptaibolin analogues by incorporation of α,α-dialkylglycines: synthesis and study of their membrane permeating ability

Analogues of Peptaibolin, a peptaibol with antibiotic activity, incorporating α,α-dialkylglycines (Deg, Dpg, and Ac6c) at selected positions were synthesised by MW-SPPS and fully characterized. A control analogue incorporating L-alanine was also prepared. The native peptide and the analogues were studied by fluorescence spectroscopy for their membrane permeating activity. Small unilamellar vesicles (SUVs) of egg phosphatidylcholine/ cholesterol (70:30) containing an encapsulated fluorescence probe (6-carboxyfluorescein) were used as membrane models. The assays of carboxyfluorescein release from SUVs upon peptide addition showed that Peptaibolin-Dpg and Peptaibolin-Ac6c are the most active peptides. These results indicate that the structure of the α,α-dialkylglycines is crucial for the membrane permeating ability of these Peptaibolin analogues.

Synthesis of polyhydroxylated compounds from Derythrose: enzymatic inhibition studies

Dissertação de mestrado em Química Medicinal

Synthesis of five-membered heterocycles by indirect electrochemical approach in "Green" media

Radical cyclization continues to be a central methodology for the preparation of natural products containing heterocyclic rings. Hence, some electrochemical results obtained by cyclic voltammetry and controlled-potential electrolysis in the study of electroreductive intramolecular cyclization of ethyl (2S, 3R)-2-bromo-3-propargyloxy-3-(2’,3’,4’,6’-tetra-O-acetyl-beta-D-glucopyranosyloxy) propanoate (1a), 2-bromo-3-allyloxy-3-(2’,3’,4’,6’-tetra-O-acetyl-beta-D-glucopyranosyloxy)propanoate (1b), 2-bromo-[1-(prop-2-yn-1-yloxy)propyl]benzene (1c) and [1-bromo-2-methoxy-2-(prop-2’-yn-1-yloxy)ethyl]benzene (1d) promoted by (1,4,8,11-tetramethyl-1,4,8,11-tetraazacyclotetradecane)nickel(I), [Ni(tmc)]+, electrogenerated at glassy carbon cathodes in ethanol and ethanol:water mixtures containing tetraalkylammonium salts, are presented. During controlled-potential electrolyses of solutions containing [Ni(tmc)]2+ and bromoalkoxylated compounds (1) catalytic reduction of the latter proceeds via one-electron cleavage of the carbon–bromine bond to form a radical intermediate that undergoes cyclization to afford the substituted tetrahydrofurans.

Potencial de produção de biodiesel com espécies oleaginosas nativas de Rondônia, Brasil

RESUMOO plano de utilização sustentável de Produtos Florestais, especialmente as espécies oleaginosas nativas tem como função a manutenção da floresta, a preservação do meio ambiente e a geração de renda em comunidades rurais. Neste sentido, este trabalho teve como objetivo avaliar a potencialidade de produção de biodiesel a partir de espécies oleaginosas nativas do estado de Rondônia. Para a identificação das espécies oleaginosas foram selecionadas cinco unidades amostrais em áreas com mata primária com o auxílio do método da Área Fixa. A extração do óleo das espécies oleaginosas foi realizada e aquelas com maior teor de lipídeos foram usadas para a produção de biodiesel. Os resultados mostraram que as espécies andiroba (Carapa guianensis), babaçu (Orbignya phalerata), castanha do Brasil (Bertholletia excelsa), tucumã-do-Amazonas (Astrocaryum tucuma) possuem potencial de produção de biodiesel. Especialmente o tucumã-do-Amazonas, por seu alto teor de lipídios (21,5%) e a qualidade do biodiesel produzido. Além disso, já havia um comércio para esta oleaginosa na região. Entretanto, torna-se imprescindível o estudo da cadeia produtiva destas espécies como estratégia para minimização na perda da cobertura vegetal nativa e na manutenção da função ambiental das áreas a serem exploradas.