Breast cancer is a leading cause of death among women in many countries. In the treatment of the breast cancer cytotoxic drugs (chemotherapy) are often used. Interindividual differences of drug response are an important cause of treatment failures. Bioavailability also depends on a major extent from the expression and activity of drug transport across biomembranes. In particular efflux transporters of the ATP-binding cassette family such as ABCB1, ABCC1 and ABCC2 have been identified as major determinants of chemoresistance in tumor cells. It was hypothesized that variance in the gene expression of membrane transporters and their genetic variance could explain at least in part interindividual differences of pharmacokinetics and clinical outcome of a variety of drugs. This thesis focuses on the functional significance of gene expression of ABCB1, ABCC1 and ABCC2 and single nucleotide polymorphisms in ABCC1 gene.

Protozoan parasites of the genus Plasmodium are the causative agent of malaria. The most severe form of human malaria is caused by P. falciparum, responsible for approximately three quarters of a million deaths each year. One major problem in the treatment of malaria is resistance to existing chemotherapies. Consequently, there is an urgent need to identify and validate novel drug targets. A possible recently identified drug target is the PfNitA protein of P. falciparum which contains orthologues in other Plasmodium species but is absent from humans. The gene is annotated as a putative formate-nitrite transporter and orthologues are found in a range of prokaryotes as well as the lower eukaryotes algae and fungi. To determine the biological function of the protein, pfnita was expressed in Escherichia coli strains lacking the endogenous formate and nitrite transporters. In order to analyse the essentiality of the gene a reverse genetics approach was taken and the data discussed. Results indicate that the PfNitA protein is located in the plasma membrane and digestive vacuole of intraerythrocytic parasites suggesting a role in the uptake or excretion of metabolites. A second complexity with regard to treatment is the lack of a vaccine. A problem in crating a vaccine is antigenic variation. The PIR family of proteins contain a so-called hypervariable domain that has led to the suggestion that the family may play a role in antigenic variation. The objective of the work carried out in this thesis was to investigate the topology and structure of the PcCir2 protein of Plasmodium chabaudi, using E. coli as the expression host. The topology of Cir2 has been examined by means of reporter fusions and overexpression/purification studies undertaken towards crystallisation. As the PcCir2 amino acid sequence does not show significant homology to other proteins, structural data may provide insights into potential functional or binding domains.

A disponibilidade de beta-alanina é o fator limitante para a síntese intramuscular de carnosina. Dessa maneira, aumentar a disponibilidade de beta-alanina para o músculo esquelético é a estratégia mais eficaz para aumentar o conteúdo de carnosina muscular. Postula-se que o transportador de beta-alanina (TauT) possa ser estimulado pela insulina. Para testar essa hipótese, examinamos se a captação de beta-alanina pelo músculo esquelético de humanos é influenciada pela hiperinsulinemia, controlando as concentrações de insulina e beta-alanina no plasma através de infusão intravenosa aguda de beta-alanina. Realizamos um estudo crossover e contrabalanceado em 12 homens jovens e saudáveis (27,5±5,1 anos). Os participantes compareceram ao laboratório em duas ocasiões separadas por 10 semanas de whashout. A beta-alanina foi infundida por via intravenosa em ambos os ensaios por 150 min a uma taxa de 0,11 g.kg.min-1. Em um ensaio, a técnica de clamp euglicêmico hiperinsulinêmico foi usada para obtermos concentrações elevadas de insulina (AI), enquanto que no outro ensaio, foram mantidas concentrações de insulina em jejum (BI). Antes e 30 minutos após a infusão de beta-alanina, amostras de músculo (biópsias percutâneas) foram coletadas para determinar o conteúdo de beta-alanina e carnosina. Coletas sanguíneas foram realizadas antes (0), 10, 30, 60, 90, 120, 150 e 30 min (180) após a infusão para análise de insulina e beta-alanina plasmáticas. Urina 24 h foi coletada após o período de infusão para análise de beta-alanina. Não houve diferenças significantes entre os ensaios na concentração de beta-alanina plasmática (p=0,20), de beta-alanina muscular (p=0,72), de carnosina muscular (p=0,82) e de beta-alanina urinária (p= 0,92). A hiperinsulinemia não aumentou a captação de beta-alanina para o músculo esquelético, nem aumentou a retenção de beta-alanina corporal, pelo menos quando as concentrações de beta-alanina excederam a Vmax do TauT. Nossas descobertas sugerem que as estratégias de suplementação de beta-alanina que manipulam as concentrações de insulina provavelmente apresentam relevância clínica limitada / Beta-alanine availability is limiting for the intramuscular synthesis of carnosine. Thus, increasing beta-alanine availability to skeletal muscle is the most effective strategy to increase muscle carnosine content. It has been postulated that the transmembrane transporter of beta-alanine (TauT) could be stimulated by insulin. To test this hypothesis, we examined whether the beta-alanine uptake by human muscle is influenced by hyperinsulinemia by controlling both insulin and beta-alanine concentrations in plasma via intravenous infusion of beta-alanine. We conducted a counterbalanced crossover study in 12 young men (27.5 ± 5.1 yr). Participants attended to the laboratory on two separated occasions, 10 weeks apart. beta-alanine was intravenously infused on both trials for 150 min at a rate of 0.11 g.kg.min-1. In one trial, a hyperinsulinemic-euglycemic clamp was used to main high insulin concentrations (HI) whereas fasting insulin concentrations (LI) was maintained in the other trial. Before and 30 min after infusion, muscle samples (percutaneous biopsies) were taken to determine beta-alanine and carnosine content. Blood samples were taken before (0), 10, 30, 60, 90, 120, 150 e 30 min (180) after the infusion for plasma insulin and beta-alanine analysis. 24 h urine was colleted after infusion for beta-alanine analysis. No significant differences in plasma beta-alanine (p=0.20), muscle beta-alanine (p=0.72), muscle carnosine (p=0.82) and urinary beta-alanine (p=0.92) were shown between conditions. Hyperinsulinemia did not increase beta-alanine uptake to muscle tissue and bodily tissues, nor did it increase whole-body beta-alanine retention, at least when beta-alanine concentrations exceed the Vmax of TauT. Our findings suggest that beta-alanine supplementation strategies that maniupulate insulin concentrations are probably of limited clinical relevance

Les interneurones GABAergiques dans l’hippocampe forment de petites populations diverses de neurones inhibiteurs contrôlant le transfert d’informations dans de larges ensembles de cellules principales. Ils compensent leur infériorité numérique par de vastes arborisations axonales capables de maintenir une libération vésiculaire de GABA à haute fréquence, et d’ajuster précisément la balance entre excitation et inhibition pour différents régimes d’activité du réseau. Les petites synapses centrales contiennent un nombre limité de vésicules synaptiques dont le recyclage par endocytose est essentiel au maintien de la transmission pendant une activité répétée. Le remplissage en GABA de ces vésicules recyclées est dépendant d’un approvisionnement des terminaisons en GABA suffisant pour faire face à la demande de recyclage créée par l’activité. Nos résultats mettent en évidence de nouveaux mécanismes d’adaptation de l’approvisionnement aux besoins imposés par le recyclage vésiculaire selon le régime d’activité, ainsi qu’un couplage direct entre le cycle de neurotransmetteurs et le cycle vésiculaire. Nous montrons que les transporteurs de glutamine sont responsables d’une potentialisation de l’approvisionnement des varicosités en GABA lors d’une activité répétée, probablement par une augmentation du nombre de ces transporteurs à la membrane. En développant et en utilisant des paradigmes expérimentaux nouveaux, nous montrons que la régulation métabolique du cycle vésiculaire passe par une adaptation du pool de vésicules recyclantes à la disponibilité en neurotransmetteurs. La nature du senseur de cette régulation et sa localisation cytosolique ou luminale restent à déterminer / In the hippocampus, GABAergic interneurons represent only 10% of the neuronal population but are able to synchronize the activity of large neuronal networks. They compensate their numerical inferiority by a large axonal arborization to sustain synaptic activity at high frequency and adjust the balance between excitation and inhibition for different regime of activity. Since small central synapses contain a limited pool of vesicles, their recycling by endocytosis is essential to maintain transmission during repeated activity. The filling of recycling vesicles with GABA is dependent on its supply in terminals which should be adjusted to the demand imposed by vesicular recycling. Our results reveal new transporter mechanisms that adapt GABA supply to neuron activity, suggesting a direct coupling between the neurotransmitter and the vesicle cycles. We show that high recycling activity increases GABA supply, probably by increasing the number of glutamine transporters at the membrane. By developing and using FM5-95 and VGAT-pHluorin with a new experimental paradigm, we provide evidence for a metabolic regulation of the vesicle cycle that involves a dynamic adaptation of the recycling pool to the neurotransmitter availability. The nature of the sensor of this regulation and its cytosolic or luminal location remain to be determined.

Rapporten undersöker skillnaden mellan vägtransporter och intermodala transporter. Detta görs bland annat genom en totalkostnadsanalys vilket undersöker kostnader relaterade till transportprocessen och även miljöpåverkan genom utsläpp de olika transportmetoderna bidrar med och diskuterar även kringliggande faktorer som påverkar valet av transportmetod. Det visar sig att vägtransporter är det billigare alternativet men de bidrar även med mycket större miljöpåverkan än Intermodala transporter.

Denna rapport innehåller en fallstudie av företaget HKScan och fokuserar på deras interna hantering av transporter ut till kunderna. Eftersom HKScan verkar i en konkurrensutsatt bransch med små marginaler så är det viktigt att varenda krona används på ett bra sätt. Transporter är en aktivitet som sällan undersöks eftersom det är en operativ aktivitet som är fristående från övrig verksamhet. Det har dock visat sig att även om olika funktioner i ett företag traditionellt styrs separat så finns det starka kopplingar mellan transporter och funktioner som sälj, marknad, inköp, ekonomi, personaladministration och produktion. Denna studies syfte var därför att undersöka vilka faktorer som påverkar transportkostnaderna och ge förslag på hur transportkostnaderna på sikt kan minskas. Undersökningen av vilka faktorer som påverkar transportkostnaderna genomfördes i flera steg. Först sammanställdes de faktorer som nämns i teorin vilket kompletterades med de faktorer som Supply Chain-funktionen på HKScan trodde påverkade transportkostnaderna. Detta resulterade i en modell med 18 möjliga påverkande faktorer som sedan avgränsades beroende på vad HKScan mäter och vad som ansågs relevant att undersöka. Denna avgränsade lista bestod av sju drivare som sedan undersöktes med hjälp av en multipel regressionsanalys. I denna analys kunde det matematiskt bevisas att faktorerna vikt per transportkolli, transporttemperatur, kvantitet, avstånd samt vilken typ av kund som transporten gick till påverkar transportkostnaden både på sändningsnivå och per transportkolli. Efter att dessa drivare identifierats så hölls intervjuer för att undersöka vilka funktioner som har störst möjlighet att påverka dem och därför i förlängningen även kan påverka transportkostnaden direkt. Dessa intervjuer visade att det främst är funktionerna för sälj och Supply Chain som direkt kan påverka transportkostnaden. Även marknad och produktion har möjlighet att påverka transportkostnaden på ett indirekt sätt. Slutligen genomfördes intervjuer med personal från de berörda funktionerna för att undersöka på vilket sätt medvetenheten och styrningen av transportkostnaderna kan förbättras. Detta resulterade i ett stort antal förslag där det mest frekvent förekommande var en kontinuerlig uppföljning av transportkostnaderna. Denna uppföljning ska inte bara vara på nivån att transportkostnaderna varit dyra utan gå djupare och peka på vilka mönster i beställningar och transporter som gett upphov till den högre kostnaden. Om denna uppföljning ska ske manuellt eller med hjälp av ett automatiserat verktyg finns det olika åsikter om men personalen känner genomgående de behöver öka medvetenheten om vad som påverkar transportkostnaderna för att kunna sänka dem. / This report contains a case study of the company HKScan and focuses on their internal handling of transports to customers. As HKScan operates in a competitive industry with small margins, it is important that all money is used in the best way. An activity that costs money but rarely is investigated is the transportation since it is seen as a very operative activity. It has, however, been found that although different functions of a company are traditionally controlled separately, there are strong links between transports and functions such as sales, marketing, purchasing, finance, human resource management and production. The purpose of this study was therefore to investigate which factors that affect transport costs and provide suggestions on how transport costs can be reduced. The study of which factors that affect transport costs was carried out in several stages. First, factors mentioned in the theory were compiled and complemented by the factors that the Supply Chain department at HKScan believed could affect the cost. This resulted in a list of 18 possible influencing factors that were then demarcated depending on what HKScan measures and which was considered relevant to investigate. This demarcated list consisted of seven factors, which were then investigated using a multiple regression analysis. From this analysis, it could be mathematically proven that the parameters quantity, distance, weight per transport package, temperature, and the type of customer to which the transport went, affects the transport cost both at the shipping level and per transport package. After these drivers had been identified, interviews were held to investigate which departments have the greatest potential to influence them and, in the long run, affect transport costs. These interviews showed that it was primarily the sales and Supply Chain departments that affect the shipping costs, even though market and production have a more indirect impact. Finally, interviews were conducted with staff from the relevant departments to investigate how awareness and management of transport costs can be improved. This resulted in a large number of proposals where the most frequent one mentioned was a continuous follow-up on transport costs. This follow-up should point out which patterns in orders and shipments that gave rise to the higher cost. If this follow-up is to be done manually or by means of an automated tool, there are different views, but the staff feels that in order to reduce transport costs, they need increased awareness of what affects them. / <p>Observera att alla siffror och resultat i rapporten är modifierade och speglar inte det verkliga resultatet.</p>

Benznidazol (BZ), fármaco utilizado para o tratamento da doença de Chagas, apresenta eficácia limitada na fase crônica da doença. Falhas terapêuticas foram atribuídas majoritariamente a diferenças na suscetibilidade a BZ entre as cepas do T. cruzi. Resultados prévios de nosso grupo indicam que o gene de um transportador ABC da subfamília G, TcABCG1, encontra-se super-expresso em cepas resistentes a BZ. Transportadores ABCG foram associados a resistência a drogas em vários organismos. O objetivo central do presente estudo foi caracterizar o gene TcABCG1 em cepas de diferentes linhagens e cuja suscetibilidade a BZ foi definida. A sequência do gene TcABCG1 (1.998 pb) de 14 cepas foi determinada. Observamos algumas variações de aminoácidos na proteína ABC entre as cepas. Análises genealógicas de TcABCG1 definiram quatro clados (TcI, TcII, TcIII e Tcbat). Os dois haplótipos das cepas híbridas TcV e TcVI agruparam com os clados TcII e TcIII. Dados de imunofluorescência indireta em epimastigotas indicam que TcABCG1 está localizado em vesículas intracelulares. / Benznidazole (BZ), drug employed for Chagas disease treatment, has limited efficacy in the chronic phase of the disease. Treatment failures have been attributed mostly to differences in BZ susceptibility among T. cruzi strains. Previous data from our group indicate that one ABC transporter gene of the G subfamily, named TcABCG1, is overexpressed in BZ-resistant strains. ABCG transporters have been associated to drug resistance in several organisms. The central goal of the present study was to characterize TcABCG1 gene in strains belonging to different lineages and of defined BZ susceptibility. TcABCG1 gene sequence (1,998 bp) of 14 strains was determined. Few amino acid substitutions were detected in the ABC transporter protein among the strains. Genealogic analyses of TcABCG1 showed four distinct clades (TcI, TcII, TcIII and Tcbat). The two haplotypes of TcV and TcVI hybrid strains clustered with TcII and TcIII clades. Indirect immunofluorescence analysis in epimastigote forms indicated that TcABCG1 is localized to intracellular vesicles.

A Proteína Transportadora de Poliaminas (PotD) é um antígeno importante para a virulência de Streptococcus pneumoniae in vivo, capaz de proteger camundongos imunizados contra infecção sistêmica, além de reduzir a colonização da nasofaringe dos animais. Porém, visando ampliar a cobertura vacinal, a combinação com outros antígenos da bactéria se faz necessária. Este trabalho teve como objetivo aprofundar o estudo sobre a resposta imune gerada contra a proteína PotD, sozinha ou em fusão com duas outras proteínas pneumocócicas: o derivado de Pneumolisina, PdT, e a proteína de superfície de pneumococo A (PspA). Para tanto, os genes potD, pdT e pspA foram clonados e expressos, sozinhos ou fusionados, gerando as proteínas híbridas rPotD-PdT e rPspA-PotD. As proteínas recombinantes e os híbridos foram utilizados na imunização subcutânea de camundongos BALB/c, gerando elevados níveis de anticorpos. O soro dos animais imunizados foi capaz de reconhecer e se ligar à superfície de diferentes isolados de pneumococos, e de ampliar a fagocitose da bactéria por células peritoneais murinas in vitro. Em todos os ensaios, os híbridos se mostraram mais eficazes do que as proteínas isoladas, induzindo anticorpos capazes de potencializar a fagocitose dos pneumococos. A resposta imune celular foi caracterizada pela produção de INF-&#947;, IL-2 e IL-17 pelos esplenócitos, e um aumento na produção de NO pelos fagócitos peritoneais dos animais imunizados. Apesar dos resultados promissores in vitro, a proteína rPotD-PdT não foi capaz de induzir proteção em nenhum dos modelos avaliados; em contraste, a fusão rPspAPotD foi capaz de proteger os camundongos contra sepse por dois isolados virulentos de pneumococo, além de reduzir a colonização na nasofaringe. Por fim, demonstramos que a adição das poliaminas transportadas por PotD, espermidina e putrescina, à cultura de pneumococos interfere na formação de biofilme in vitro. Cnsiderando o importante papel da formação de biofilmes na colonização, este resultado sugere um possível mecanismo de ação da PotD durante a colonização por pneumococo. Em conjunto, os resultados deste estudo sugerem que a utilização de uma formulação híbrida, rPspA-PotD, compreende uma estratégia vacinal promissora, capaz de proteger contra colonização e sepse pneumocócica, pela produção de anticorpos opsonizantes e ativação de citocinas protetoras, como IL-17. / Polyamine Transporter D (PotD) is an important antigen for Streptococcus pneumoniae virulence in vivo, protecting immunized mice against systemic infection and reducing the bacterial load in the nasopharynx of immunized animals. However, in order to extend vaccine coverage, the combination of PotD with other antigens of the bacterium is required. The present study aimed at expanding the investigation of the immune response generated against PotD alone or fused with two other pneumococcal proteins: the Pneumolysin derivative, PdT and Pneumococcal Surface Protein A (PspA). Therefore, the potD, pdt and pspA genes were cloned and expressed, either alone or in fusion, generating the hybrid proteins rPotD-PdT and rPspA-PotD. The recombinant proteins and hybrids were used for subcutaneous immunization of BALB/c mice, generating high levels of antibodies. Sera from immunized animals were able to recognize and bind onto the surface of different pneumococcal strains, and to enhance phagocytosis of the bacterium in vitro. In all tests, the hybrids were more effective than the isolated proteins. The cellular immune response was characterized by the production of INF-&#947;, IL-2 and IL-17 by splenocytes and increased production of NO by peritoneal cells of the immunized animals. Despite promising results in vitro, rPotD-PdT protein was not able to induce protection in any of the tested challenge models. In contrast, rPspA-PotD fusion was able to protect mice against sepsis with two virulent isolates of pneumococcus and led to reduction in bacterial loads in the nasopharynx of challenged animals. Finally, we demonstrate that the addition of exogenous polyamines, spermidine, and putrescine, in the pneumococcal culture interfered with biofilm formation in vitro. Considering the important role of biofilm formation for successful colonization, this result suggests a possible mechanism of action of PotD during colonization by pneumococcus. Taken together, the results suggest that the use of the hybrid rPspA-PotD comprises a promising vaccine strategy, able to protect against colonization and pneumococcal sepsis, through the production of opsonizing antibodies and activation of protective cytokines, such as IL-17.

Au cours des dernières décennies, une augmentation non négligeable du phénomène de résistance aux antibiotiques des bactéries a été observée. Ces bactéries possèdent plusieurs mécanismes de résistance parmi lesquels l’utilisation de transporteurs de type MDR (MultiDrug Resistance) dont certains appartiennent à la famille des transporteurs ABC (ATP-Binding Cassette). Les transporteurs ABC sont des protéines membranaires et ubiquitaires qui possèdent une topologie commune avec deux domaines transmembranaires et deux domaines cytoplasmiques. Les transporteurs ABC de type exportateur permettent le transport de molécules à l’extérieur de la cellule en utilisant l’énergie fournie par l’hydrolyse de l’ATP. PatA-PatB est un transporteur ABC de Streptococcus pneumoniae, un pathogène humain responsable de pneumonies et de méningites. Cette protéine est impliquée dans la résistance de ce pathogène à des antibiotiques de types fluoroquinolones. Pour étudier son mécanisme moléculaire, nous avons optimisé la surexpression fonctionnelle de ce transporteur chez Escherichia coli. Ainsi, nous avons pu caractériser son activité de transport de drogues et son activité d’hydrolyse de nucléotides. Ces expériences ont révélé que PatA-PatB a la particularité d’utiliser préférentiellement le GTP comme source d’énergie, contrairement aux autres membres de cette famille. Afin d’identifier l’origine de cette propriété au niveau moléculaire, des expériences de mutagénèse dirigée ont été effectuées et nous avons ainsi identifié deux simples mutants qui transportent les drogues aussi bien avec du GTP que de l’ATP / The excessive use of antibiotics during the past decades led to the amplification of multidrug resistance in pathogenic bacteria. Bacteria have developed several mechanisms of antibiotic resistance. One of them involves the antibiotic efflux by MDR (MultiDrug Resistance) transporters, some of which belong to the ABC (ATP-Binding Cassette) transporter family. ABC transporter are ubiquitous membrane proteins with a conserved topology comprising four domains : two «TransMembrane Domain» and two cytoplasmic domains named « Nucleotide-Binding Domain ». ABC exporters expel drugs outside the bacteria using the energy of ATP hydrolysis. PatA-PatB is an ABC transporter from Streptococcus pneumoniae, a human pathogen bacterium responsible for pneumonia and meningitis. This protein is involved in S. pneumoniae resistance against fluoroquinolone antibiotics. To study the molecular mechanism, we optimized the functional expression of this transporter in Escherichia coli. Then, we characterized its drug transport activity and its nucleotide hydrolysis activity. These experiments showed that PatA-PatB, in contrast to other members of the ABC superfamily, preferentially uses GTP as energy supply. To identify the origin of this property at a molecular level, mutagenesis experiments were performed and we identified two mutants capable of an even drug transport with ATP and GTP

O presente trabalho teve por objetivo avaliar o perfil metabólico da utilização dos açúcares por diferentes leveduras (BG-1, CAT-1, FLEISCHMANN, PE-2, 1403-7A e LCM001) em diferentes meios de crescimento e também analisar o comportamento e a atividade de invertase destas leveduras durante um processo fermentativo com reciclos de células, semelhantemente ao processo industrial, levando em consideração os parâmetros: produção de etanol, formação de biomassa, produção de trealose, glicogênio, glicerol, rendimento e eficiência fermentativa. Os experimentos foram divididos em três partes: a 1a foi a quantificação da atividade de invertase das diferentes leveduras, em mosto de fermentação à base de mel e água (13% ART), a 2a analisouse o perfil de metabolização em diferentes meios de crescimento à base de YEP com 2% de açúcares (glicose, sacarose, ou glicose + sacarose), e a 3a foi a realização de uma fermentação alcoólica com 4 reciclos de células, com mosto de mel (13% ART), sendo os três primeiros utilizados para a avaliação do rendimento fermentativo, bem como seus indicadores (trealose, glicogênio, viabilidade, entre outros) e o último, destinado ao perfil de metabolização de açúcares em condições de fermentação. Os resultados mostram que as leveduras possuem diferentes atividades de invertase (BG-1 = 7,34; FLEISCHMANN = 5,75; CAT-1 = 3,76; PE-2 = 2,39 gART.h-1.gbiomassa; 1403-7A e LCM001, não possuem atividade), apresentam diferentes velocidades de hidrólise da sacarose, tanto em meios de crescimento como mostos (BG-1 e FLEISCHMANN = 2 h; CAT-1 = 3h; PE-2 = 4h; 1403-7A = 24 h e LCM001 = >24 h), assim como a velocidade de metabolização dos açúcares presentes nestes. Conclui-se que a atividade de invertase é dependente da linhagem de levedura, assim como a velocidade de metabolização dos açúcares em meios de crescimento e mosto. A análise do perfil de metabolização de açúcares não permite identificar a presença de transportador de sacarose. No geral, as leveduras selecionadas apresentam melhor desempenho fermentativo. Por outro lado, verificou-se que as linhagens com transportador de sacarose apesar da menor produção de álcool, apresentaram uma boa eficiência fermentativa. O maior problema por parte destas últimas, é o alto tempo de fermentação e a baixa taxa de multiplicação, com conseqüente queda na viabilidade celular. / The aim of this work was to evaluate the metabolic profile in the utilization of sugars, using different yeast strains (BG-1, CAT-1, FLEISHMANN, PE-2, 1403-7A and LCM001) with different growth medium and also to analyze behavior and the invertase activities of these yeast strains during fermentative process with recycling, similar to the industrial process. The following parameters were used: ethanol production, biomass formation, trealose production, glycogen, glycerol, ethanol yield and fermentation efficiency. The trials were divided into three parts: First was activity quantification of invertase using different yeast strains, with a mix of molasses and water (13% ART), the second was analyzed the metabolization profile with different growth medium using YEP added 2% of sugars (glucose, sucrose, or glucose + sucrose), and the third it was the alcoholic fermentation with 4 yeast cell recycles, using wort of molasses and juice ( 13 % ART), The first three were utilized to evaluate the fermentation yield and also theirs indicatives (trealose, glycogen, viability within others) and, the last one was performed to study the profile of sugars metabolization in fermentation conditions. The results showed that those yeasts produced different invertase activities (BG-1 = 7,34; FLEISCHMANN = 5,75; CAT-1 = 3,76; PE-2 = 2,39 gART.h-1.gbiomass; 1403-7A and LCM001 did not have invertase activity), different velocity of sucrose hydrolysis, as much as medium growth as worts (BG-1 and FLEISCHANN = 2 h; CAT-1 = 3h; PE-2 = 4h; 1403-7A = 24 h and LCM001 = >24 h), also as the velocity of sugars metabolization present on this medium. It can be concluded that the invertase activity is dependent of yeast strain, as the velocity of sugars metabolization in growth medium and wort. The profile analysis of metabolization of sugars did not allow to identify the presence of sucrose transporter. In general, the selected yeasts present the best fermentation performance. On the other hand, it was observed the strains with sucrose transportation did not show stress. The major problem of these yeasts were the high fermentation time and low propagation rate, and a decrease of the viability.