Functional and molecular characterization of SR45, a plant-specific factor involved in sugar and stress signaling in Arabidopsis thaliana

Dissertation presented to obtain the Ph.D degree in Molecular Biology

A multitask ATPase serving different ABC-type sugar importers in Bacillus subtilis

Journal of Bacteriology (Out 2010) 5312-5318

Characterization and regulation of a bacterial sugar phosphatase of the haloalkanoate dehalogenase superfamily, AraL, from Bacillus subtilis

FEBS journal, Volume 278, Issue 14, pages 2511-2524, July 2011

Advances on the production of polyhydroxyalkanoates (PHA) by mixed microbial cultures from sugar cane molasses

Dissertação para obtenção do Grau de Mestre em Biotecnologia

An RBC model with a rich fiscal sector

A Work Project, presented as part of the requirements for the Award of a Masters Degree in Economics from the NOVA – School of Business and Economics

Synthesis and characterization of sugar containing hydrophilic and hydrophobic polymers with potential application in medicine

Dissertation toobtaina Master of Science degree in Bioorganics

Constraint-based modelling of mixed microbial populations: Application to polyhydroxyalkanoates production

Dissertação para obtenção do Grau de Doutor em Engenharia Química e Bioquímica

Characterization and regulation of a bacterial sugar phosphatase of the HAD superfamily, AraL, from Bacillus subtilis.

AraL from Bacillus subtilis is a member of the ubiquitous haloalkanoate dehalogenase, HAD, superfamily. The araL gene has been cloned, over-expressed in Escherichia coli and its product purified to homogeneity. The enzyme displays phosphatase activity, which is optimal at neutral pH (7.0) and 65 °C. Substrate screening and kinetic analysis showed AraL to have low specificity and catalytic activity towards several sugar phosphates, which are metabolic intermediates of the glycolytic and pentose phosphate pathways. Based on substrate specificity and gene context within the arabinose metabolic operon, a putative physiological role of AraL in detoxification of accidental accumulation of phosphorylated metabolites has been proposed. The ability of AraL to catabolise several related secondary metabolites requires regulation at the genetic level. Here, by site- directed mutagenesis, we show that AraL production is regulated by a structure in the translation initiation region of the mRNA, which most probably blocks access to the ribosome-binding site, preventing protein synthesis. Members of HAD subfamily IIA and IIB are characterised by a broad-range and overlapping specificity that anticipated the need for regulation at the genetic level. In this study we provide evidence for the existence of a genetic regulatory mechanism controlling AraL production.

Fermentation of fruit juices by the osmotolerant yeast Candida magnoliae

This study focuses on the assessment of the fermentation conditions required to modulate the metabolic flux in the osmotolerant yeast Candida magnoliae and evaluate its potential to produce low-alcoholic and low-caloric fermented beverages. For that purpose, two strains, PYCC 2903 and PYCC 3191, were used and fermentation conditions as oxygenation, sugar concentration and the ratio of glucose to fructose were studied using synthetic culture media. Candida magnoliae PYCC 2903 was subsequently used to ferment real industrial fructose-rich substrates such as fruit juices. Sugar consumption profiles for C.magnoliae PYCC 2903 incubated aerobically in the presence of high fructose and glucose concentrations (15%, 10% and 5%) showed a selective utilization of fructose, denoting a preference for this sugar over glucose. The lower ratio between ethanol and sugar alcohols yield was obtained for both strains incubated under oxygen limitation simulating industrial fructose-rich substrates, confirming the ability of this yeast to direct fermentation towards alternative products. Enzymatic assays for hexokinase activity in terms of capacity and affinity for glucose and fructose were performed, aiming to elucidate its contribution to the fructophilic behaviour of this yeast. Enzymatic assays for both strains showed that the Vmax is two to threefold higher for fructose than for glucose but Km is also 10-20-fold higher for this sugar than for glucose. Hence, hexokinase kinetic properties do not explain fructophily in C.magnoliae. This indicates that fructose transport is probably determining in this respect, as observed for other fructophilic yeasts. Fruit juice fermentations with C.magnoliae PYCC 2903 revealed a potential for the production of beverages with interesting sensorial properties. Pear and peach fermentations exhibited the best results with the lowest ratio between ethanol and sugar alcohols yield and the most pleasant organoleptic features.

Contributos para o dimensionamento de um armazém: caso de estudo na Nutriceal Foods

Atualmente, as exigências de um mercado cada vez mais consumista obrigam as empresas a melhorar o seu desempenho global de modo a conseguirem corresponder a todas as expetativas e necessidades impostas pelos clientes. É da responsabilidade das empresas, de modo a tornarem-se mais competitivas, aumentar a eficiência e eficácia da cadeia de abastecimento em que estão inseridas. A logística representa custos elevados para as empresas, nomeadamente na sua atividade de armazenagem, o que implica um maior controlo por parte dos gestores de modo a torná-la mais eficiente, representando, assim, vantagens competitivas face à concorrência. Esta dissertação visa o cálculo dos custos necessários para a abertura de um armazém externo aos terrenos da fábrica para a empresa Nutriceal Foods e o desenvolvimento de propostas de melhoria relativamente ao processo de gestão aplicado no atual armazém sub-contratado a uma empresa externa. Em primeiro lugar, foram analisados os custos da Nutriceal Foods associados à sub-contratação do armazém externo, nomeadamente os custos com a movimentação das mercadorias, transporte e armazenagem. Em seguida foram desenvolvidas duas propostas de melhoria que visam a abertura de um armazém externo, tendo sido dimensionada a área de armazenagem pretendida face à previsão do volume de paletes a armazenar em 2015, e definido um layout para cada uma das propostas alternativas. A proposta 1 reduz os custos em relação ao armazém sub-contratado em 37,5%, enquanto que a proposta 2 reduz os custos em 39,4%. De entre as propostas analisadas foi selecionada a proposta 1, por corresponder aos parâmetros pré-definidos pela empresa de minimização dos custos globais do armazém, visando o seu aluguer. Por fim, são sugeridas propostas de trabalho futuro, visando a melhoria da gestão do armazém selecionado.