Chlorite Dismutase from perchlorate reducing bacteria. A potential biocatalyst to eliminate chlorinated species from polluted water resources and industrial wastes

Magnetospirillum (M.) sp. strain Lusitani, a perchlorate reducing bacteria (PRB), was previously isolated from a wastewater treatment plant and phylogenetic analysis was performed to classify the isolate. The DNA sequence of the genes responsible for perchlorate reduction and chlorite dismutation was determined and a model was designed based on the physiological roles of the proteins involved in the pcr-cld regulon. Chlorite dismutase (Cld) was purified from Magnetospirillum sp. strain Lusitani cells grown in anaerobiosis in the presence of perchlorate. The protein was purified up to electrophoretic grade using HPLC techniques as a 140 kDa homopentamer comprising five ~28 kDa monomers. Steady-state kinetic studies showed that the enzyme follows a Michaelis-Menten model with optimal pH and temperature of 6.0 and 5°C, respectively. The average values for the kinetic constants KM and Vmax were respectively 0.56 mM and 10.2 U, which correspond to a specific activity of 35470 U/mg and a turnover number of 16552 s-1. Cld from M. sp. strain Lusitani is inhibited by the product chloride, but not by dioxygen. Inhibition constants KiC= 460 mM and KiU= 480 mM indicated that sodium chloride is a weak mixed inhibitor of Cld, with a slightly stronger competitive character. The X-ray crystallography structure of M. sp. strain Lusitani Cld was solved at 3.0 Å resolution. In agreement with cofactor content biochemical analysis, the X-ray data showed that each Cld monomer harbors one heme b coordinated by a histidine residue (His188), hydrogen-bonded to a conserved glutamic acid residue (Glu238). The conserved neighboring arginine residue (Arg201) important for substrate positioning, was found in two different conformations in different monomers depending on the presence of the exogenous ligand thiocyanate. UV-Visible and CW-EPR spectroscopies were used to study the effect of redox agents, pH and exogenous ligands on the heme environment.

Optimization of bioplastics production from cheese whey

Dissertation presented in partial fulfillment of the requirements for the degree of Master in Biotechnology

Food waste valorization through the production of polyhydroxyalkanoates by mixed microbial cultures

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

Production and characterization of Chitin-Glucan Complex by Komagataella pastoris

This thesis was focused on the production, extraction and characterization of chitin:β-glucan complex (CGC). In this process, glycerol byproduct from the biodiesel industry was used as carbon source. The selected CGC producing yeast was Komagataella pastoris (formerly known as Pichia pastoris), due the fact that to achieved high cell densities using as carbon source glycerol from the biodiesel industry. Firstly, a screening of K. pastoris strains was performed in shake flask assays, in order to select the strain of K. pastoris with better performance, in terms of growth, using glycerol as a carbon source. K. pastoris strain DSM 70877 achieved higher final cell densities (92-97 g/l), using pure glycerol (99%, w/v) and in glycerol from the biodiesel industry (86%, w/v), respectively, compared to DSM 70382 strain (74-82 g/l). Based on these shake flask assays results, the wild type DSM 70877 strain was selected to proceed for cultivation in a 2 l bioreactor, using glycerol byproduct (40 g/l), as sole carbon source. Biomass production by K. pastoris was performed under controlled temperature and pH (30.0 ºC and 5.0, respectively). More than 100 g/l biomass was obtained in less than 48 h. The yield of biomass on a glycerol basis was 0.55 g/g during the batch phase and 0.63 g/g during the fed-batch phase. In order to optimize the downstream process, by increasing extraction and purification efficiency of CGC from K. pastoris biomass, several assays were performed. It was found that extraction with 5 M NaOH at 65 ºC, during 2 hours, associated to neutralization with HCl, followed by successive washing steps with deionised water until conductivity of ≤20μS/cm, increased CGC purity. The obtained copolymer, CGCpure, had a chitin:glucan molar ratio of 25:75 mol% close to commercial CGC samples extracted from A. niger mycelium, kiOsmetine from Kitozyme (30:70 mol%). CGCpure was characterized by solid-state Nuclear Magnetic Resonance (NMR) spectroscopy and Differential Scanning Calorimetry (DCS), revealing a CGC with higher purity than a CGC commercial (kiOsmetine). In order to optimize CGC production, a set of batch cultivation experiments was performed to evaluate the effect of pH (3.5–6.5) and temperature (20–40 ºC) on the specific cell growth rate, CGC production and polymer composition. Statistical tools (response surface methodology and central composite design) were used. The CGC content in the biomass and the volumetric productivity (rp) were not significantly affected within the tested pH and temperature ranges. In contrast, the effect of pH and temperature on the CGC molar ratio was more pronounced. The highest chitin: β-glucan molar ratio (> 14:86) was obtained for the mid-range pH (4.5-5.8) and temperatures (26–33 ºC). The ability of K. pastoris to synthesize CGC with different molar ratios as a function of pH and temperature is a feature that can be exploited to obtain tailored polymer compositions.(...)

Production of bacterial biopolymers from industrial fat-containing wastes

Dissertation presented in partial fulfilment of the Requirements for the Degree of Master in Biotechnology

Lime mortars with ceramic wastes: characterization of components and their influence on the mechanical behaviour

Considering the fundamental importance of preserving the built heritage and of ensuring the good performance achieved by incorporating ceramic particles in lime mortars in ancient times, it is important to study solutions that use materials the available today, in order to produce mortars intended to repair and replace the old ones. Solutions incorporating industrial ceramic waste might be profitable for several reasons, namely for economic, environmental and technical aspects. In this paper, seven ceramic waste products collected from ceramics factories are characterized. Their mineralogy, dimensional features and pozzolanicity were determined. Three of these products, with different particle size fractions (obtained directly from milling, dust only and fragment fractions only), were selected, incorporated into air lime mortars, and their mechanical strength was determined. In the present work, evidence of mechanical efficiency, when common sand or air lime were partially replaced by ceramic wastes, was made clear, drawing attention to the sustainability of this type of mortars, hence, encouraging further research.

Estudo da eficiência da ETAR da zona industrial da AutoEuropa por análise biológica

No tratamento de águas residuais por lamas activadas, existem diferentes populações de protozoários e metazoários que se podem desenvolver durante o processo. A sua presença é fundamental para o bom desempenho da ETAR. Além de reduzirem a turvação do efluente final, estimulam o crescimento bacteriano, promovem a floculação e algumas espécies reduzem a poluição. É possível estabelecer uma estreita relação entre a predominância de determinadas espécies ou grupos de espécies, com parâmetros operacionais da ETAR, tais como os índices bióticos. Este procedimento requer a identificação e enumeração das diferentes espécies. No entanto, existem outros métodos, que apresentam relações entre a abundância de uma dada espécie ou grupo e os parâmetros da estação. O presente trabalho tem como objectivos: avaliar o funcionamento do sistema de tratamento de lamas activadas através da observação microscópica de protozoários e metazoários, identificação e contagem das espécies relevantes; estabelecer relações entre os organismos que contribuem para o processo de depuração de águas residuais e as condições de operação, como parâmetros físicoquímicos ou o desempenho da ETAR; e produzir uma ferramenta de apoio à gestão da ETAR, baseada em princípios biológicos. Pretende-se, ainda, contribuir e incentivar para a adopção de metodologias semelhantes em outros sistemas de lamas activadas em funcionamento no nosso País. Em termos metodológicos, a colheita de amostras foi realizada semanalmente na ETAR da ZIA durante dois meses e meio. Os parâmetros operacionais e físico-químicos foram obtidos junto da ETAR. A análise biológica foi efectuada por microscopia óptica. Avaliou-se qualitativa e quantitativamente a microfauna presente no tanque de arejamento, no poço de mistura de lamas e no clarificado de ambos os decantadores, a fim de se verificar as possíveis correlações desses parâmetros com a eficiência do sistema. Aplicou-se os resultados biológicos obtidos, em quatro modelos propostos para a avaliação do processo de tratamento, baseada em análises biológicas. Em termos qualitativos e na maior parte das observações, a lama apresentou boa qualidade. A ETAR da ZIA foi colonizada por tecamebas, principalmente Arcella sp. e Euglypha alveolata, em todo, o estudo e em todos os pontos analisados. No tanque de arejamento foram sempre observados ciliados nadadores livres. A densidade total de organismos no tanque de arejamento variou de 2982 e 7330 organismos/mL, com valor médio de 5995 organismos/mL. Relativamente, aos parâmetros estudados, os objectivos de remoção para o CBO5, CQO, SST e SSV foram alcançados.

Redução de ruído ambiental em área de produção industrial

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em: Instrumentação Manutenção Industrial e Qualidade

Manutenção industrial e custo do ciclo de vida - extracção oleaginosas

Dissertação apresentada na Faculdade de Ciências e Tecnologias para obtenção do grau de Mestre em Engenharia e Gestão Industrial

Análise de funções de segurança num processo industrial e num posto de transformação de energia eléctrica, numa industria papeleira, aplicando a metodologia SFA

Dissertação apresentada na Faculdade de Ciências e Tecnologia da Universidade Nova de Lisboa para obtenção do grau de Mestre em Engenharia Industrial

Estágio nas Edições Colibri. Uma editora "familiar" de produção (quase) industrial

Relatório de Estágio apresentado para cumprimento dos requisitos necessários à obtenção do grau de Mestre em Edição de Texto

Intelligent decision support systems for collaboration in industrial plants

Dissertação apresentada para obtenção do Grau de Doutor em Sistemas de Informação Industriais, Engenharia Electrotécnica, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Synthesis of biodiesel from wastes of food industry via direct transesterification with methanol/carbon dioxide mixtures

Dissertation presented at Faculdade de Ciências e Tecnologia from Universidade Nova de Lisboa to obtain the degree of Master in Chemical and Biochemical Engineering

O e-learning na manutenção industrial uma proposta de e-learning aplicada à manutenção produtiva total

Dissertação para obtenção do Grau de Mestre em Engenharia e Gestão Industrial