Purification of complex biopharmaceuticals with new processes, advanced analytics and computer-aided process design tools

Dissertação para obtenção do Grau de Mestre em Biotecnologia

Genetic and functional analysis of the bacillus subtilis BSP1 gam cluster

Mannans (linear mannan, glucomannan, galactomannan and galactoglucomannan) are the major constituents of the hemicellulose fraction in softwoods and show great importance as a renewable resource for fuel or feedstock applications. As complex polysaccharides, mannans can only be degraded through a synergistic action of different mannan-degrading enzymes, mannanases. Microbial mannanases are mainly extracellular enzymes that can act in wide range of pH and temperature, contributing to pulp and paper, pharmaceutical, food and feed, oil and textile successful industrial applications. Knowing and controlling these microbial mannan-degrading enzymes are essential to take advantage of their great biotechnological potential. The genome of the laboratory 168 strain of Bacillus subtilis carries genes gmuA-G dedicated to the degradation and utilization of glucomannan, including an extracellular -mannanase. Recently, the genome sequence of an undomesticated strain of B. subtilis, BSP1, was determined. In BSP1, the gmuA-G operon is maintained, interestingly, however, a second cluster of genes was found (gam cluster), which comprise a second putative extracellular β-mannanase, and most likely specify a system for the degradation and utilization of a different mannan polymer, galactoglucomannan. The genetic organization and function of the gam cluster, and whether its presence in BSP1 strain results in new hemicellulolytic capabilities, compared to those of the laboratory strain, was address in this work. In silico and in vivo mRNA analyses performed in this study revealed that the gam cluster, comprising nine genes, is organized and expressed in at least six different transcriptional units. Furthermore, cloning, expression, and production of Bbsp2923 in Escherichia coli was achieved and preliminary characterization shows that the enzyme is indeed a β-mannanase. Finally, the high hemicellulolytic capacity of the undomesticated B. subtilis BSP1, demonstrated in this work by qualitative analyses, suggests potential to be used in the food and feed industries.

Auto-consumo directo: definição de metodologia e estudo de caso

Actualmente uma das preocupações da Comissão Europeia passa pela maneira como a energia é convertida e o modo como esta é aproveitada. Surgiu assim a estratégia Europa 2020, onde é claramente expressa a preocupação relativamente ao Ambiente e a diminuição da pegada ecológica. No campo relativo ao Ambiente e Energia existem metas tais como a redução das emissões de gases de efeito de estufa em 20%, o aumento da eficiência energética em 20% e a obtenção de 20% da energia consumida via energias renováveis. De maneira a que seja possível atingir as metas propostas, têm de surgir novos regimes para as energias renováveis uma vez que se tem verificado uma diminuição de apoios às tarifas feed-in. Estes factos levaram ao surgimento de um novo regime, o auto-consumo directo, que em tudo se diferencia das tarifas feed-in, uma vez que neste regime o produtor vendia toda a energia que produzia à rede. Já no regime de auto-consumo directo, o produtor consome a energia que produz, passando a ser um prosumer. Nesta dissertação é feito o levantamento sobre o conceito de auto-consumo directo assim como a legislação e ponto de situação da Europa e de Portugal no que diz respeito à paridade de rede. Foram ainda definidas as metodologias a aplicar. Com o auxílio da ferramenta informática desenvolvida em ambiente Java foi feita a análise de dois estudos de caso, uma indústria e um comércio e foi estudada a viabilidade económica, de cada caso, segundo vários indicadores e parâmetros definidos.

Analysis and visualization of energy use for university campus

The reduction of greenhouse gas emissions is one of the big global challenges for the next decades due to its severe impact on the atmosphere that leads to a change in the climate and other environmental factors. One of the main sources of greenhouse gas is energy consumption, therefore a number of initiatives and calls for awareness and sustainability in energy use are issued among different types of institutional and organizations. The European Council adopted in 2007 energy and climate change objectives for 20% improvement until 2020. All European countries are required to use energy with more efficiency. Several steps could be conducted for energy reduction: understanding the buildings behavior through time, revealing the factors that influence the consumption, applying the right measurement for reduction and sustainability, visualizing the hidden connection between our daily habits impacts on the natural world and promoting to more sustainable life. Researchers have suggested that feedback visualization can effectively encourage conservation with energy reduction rate of 18%. Furthermore, researchers have contributed to the identification process of a set of factors which are very likely to influence consumption. Such as occupancy level, occupants behavior, environmental conditions, building thermal envelope, climate zones, etc. Nowadays, the amount of energy consumption at the university campuses are huge and it needs great effort to meet the reduction requested by European Council as well as the cost reduction. Thus, the present study was performed on the university buildings as a use case to: a. Investigate the most dynamic influence factors on energy consumption in campus; b. Implement prediction model for electricity consumption using different techniques, such as the traditional regression way and the alternative machine learning techniques; and c. Assist energy management by providing a real time energy feedback and visualization in campus for more awareness and better decision making. This methodology is implemented to the use case of University Jaume I (UJI), located in Castellon, Spain.

Barreiras e estímulos ao aleitamento materno exclusivo na Província do Uíge, Angola

RESUMO - É recomendado o aleitamento materno exclusivo desde o nascimento até aos 6 meses. Angola apresenta uma situação bastante preocupante, pois é um dos países do mundo com maior taxa de mortalidade infantil: em 2010 era de 114,9 mortes por cada 1000 nascimentos. O desmame precoce, a contaminação dos alimentos e da água potável e a desnutrição criam condições para aparecimento de doenças que causam elevadas mortes em crianças com menos de um ano. Em muitas regiões de África, o parto ainda é um acontecimento marcado por práticas culturais ancestrais que podem prejudicar a saúde e sobrevivência dos recém-nascidos, além de pôr em perigo a vida das mães. Algumas dessas práticas incluem: (i) deitar fora o colostro; e (ii) alimentar os bebés com outros alimentos que não o leite materno. Este estudo é quantitativo com metodologia transversal e pretende identificar, descrever e analisar os fatores que contribuem para o abandono do aleitamento materno exclusivo antes dos 6 meses de idade, na província do Uíge, em Angola, bem como indicar formas de intervenção para que os índices de prevalência do aleitamento materno exclusivo possam aumentar. A população em estudo são mães com filhos até um ano de idade e utilizadoras do centro de saúde materno infantil do município sede da província de Uíge em Angola, da qual será considerada uma amostra de 418 mães. Os dados obtidos através de questionário serão registados em quadros e gráficos para posterior análise estatística através de tabelas de frequências, cálculo de percentagens e taxas de incidência.

Functional characterization and directed engineering of redox proteins for a rational improvement of Bioelectrochemical systems

In recent years, new methods of clean and environmentally friendly energy production have been the focus of intense research efforts. Microbial fuel cells (MFCs) are devices that utilize naturally occurring microorganisms that feed on organic matter, like waste water, while producing electrical energy. The natural habitats of bacteria thriving in microbial fuel cells are usually marine and freshwater sediments. These microorganisms are called dissimilatory metal reducing bacteria (DMRB), but in addition to metals like iron and manganese, they can use organic compounds like DMSO or TMAO, radionuclides and electrodes as terminal electron acceptors in their metabolic pathways.(...)

Produção de electricidade em Portugal

In the present work we intend to do an analysis of the production of electricity in special regime in Portugal. We will focus in particular in the remuneration system through the feed-in tariffs. First, we will excurse throughout different legal diplomas that regulated the special regime in Portugal, exploring which guarantees were conferred to electricity generators throughout the years. We intend to also evaluate how the producers remunerative rights were (or not) protected in the various legislative changes. In the second part of the dissertation we will examine whether the feed-in tariffs may be considered as State aid. Due to the inclusion of the subject in EU Law, we will analyze EU regulation and case law to support our position about the Portuguese regime. Finally, and to the extent that the production of electricity in special regime has undergone several changes to its remunerative regime in the last few years, we propose to analyze more carefully the amendments in question. We will scrutinize the reasons that based the amendments in question, which are mainly based on the economic crisis suffered by the country. We will also examine how those changes may jeopardize the remunerative rights of the producers.

Towards modification of Medicago truncatula epigenome: genome editing with engineered nucleases

Periodic drought is the primary limitation of plant growth and crop yield. The rise of water demand caused by the increase in world population and climate change, leads to one of the biggest challenges of modern agriculture: to increase food and feed production. De novo DNA methylation is a process regulated by small interfering RNA (siRNAs), which play a role in plant response and adaptation to abiotic stress. In the particular case of water deficit, growing evidences suggest a link between the siRNA pathways and drought response in the model legume Medicago truncatula. As a first step to understand the role of DNA methylation under water stress, we have set up several bioinformatics and molecular methodologies allowing the design of Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems and the assembly of TALENs (transcription activator-like effector nucleases), to target both dicer-like 3 (MtDCL3) and RNA-Dependent RNA polymerase (MtRDR2), enzymes of the RNA-directed DNA methylation pathway. TALENs efficiency was evaluated prior to plant transformation by a yeast-based assay using two different strategies to test TALENs activity: Polyacrylamide gel electrophoresis (PAGE) and Single strand conformation polymorphisms (SSCP). In this assay, yeast cells triple transformation emerged as good and rapid alternative to laborious yeast mating strategies. PAGE analysis might be a valuable tool to test TALENs efficacy in vivo if we could increase TALENs activity. SSCP-based approach proved to be ineffective due to the generation of several false positives. TALENs and CRISPR/Cas9 system constructed and designed in this work will in the future certainly enable the successful disruption of DCL3 and RDR2 genes and shed the light on the relationship between plant stress resistance and epigenetic regulation mediated by siRNAs in M.truncatula.

Using artificial neural networks to generate trading signals for crude oil, copper and gold futures

In this thesis, a feed-forward, back-propagating Artificial Neural Network using the gradient descent algorithm is developed to forecast the directional movement of daily returns for WTI, gold and copper futures. Out-of-sample back-test results vary, with some predictive abilities for copper futures but none for either WTI or gold. The best statistically significant hit rate achieved was 57% for copper with an absolute return Sharpe Ratio of 1.25 and a benchmarked Information Ratio of 2.11.