Programa de Pós-Graduação em Biotecnologia - PPGBIOTEC/ICB
URI Permanente desta comunidadehttps://repositorio.ufpa.br/handle/2011/6089
O Programa de Pós-Graduação em Biotecnologia (PPGBIOTEC) é um programa do Instituto de Ciências Biológicas (ICB) da Universidade Federal do Pará (UFPA). O PPGBIOTEC fornece a base teórica para o desenvolvimento de projetos em duas áreas de concentração e quatro linhas de pesquisa, definidas de acordo com a competência e experiência de orientação dos docentes do programa: Área de Bioprospecção: linhas de pesquisa: (i) química orgânica de biomoléculas e metabolômica; e (ii) genômica, transcriptômica, proteômica e bioinformática de microrganismos e plantas (Bactéria, Archaea e Eukarya). Área de Biotecnologia Aplicada: linhas de pesquisa: (i) saúde humana e veterinária; e (ii) produção de insumos e processos industriais.
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Item Acesso aberto (Open Access) Estudo da combinação de preparações enzimáticas no rendimento de extração aquosa do óleo da polpa de Euterpe oleracea Martius(Universidade Federal do Pará, 2016-03-23) FERREIRA, Erika de Souza; HERMAN, Christelle Anne Nicole Paule; http://lattes.cnpq.br/8386417434654533The oil of the açai palm tree (Euterpe oleracea) fruits has a beneficial lipid profile for health with a high content of mono and polyunsaturated fatty acids, besides antioxidant substances, which provides its appreciation on the industry. The aqueous extraction of vegetable oils supported by enzymes has been widely used because of its benefits compared to the traditional processes. In this context, this work aims to identify the best combination of enzymatic preparations, which would maximize the aqueous extraction yield of the oil of the açai palm tree fruits from the pulp. The açai pulp was obtained from fruits collected in Abaetetuba during the 2015 crop. Four commercial enzymatic preparations (Novozymes) were used: Celluclast 1,5L (cellulase); Viscozyme L (endo 1,3(4) β-glucanase, xylanase, cellulase and hemicellulase); Ultrazym AFP (pectin lyase, cellulase and polygalacturonase); and Shearzyme 500L (endo 1,4-xylanase). The tests of açai oil extraction were performed by a process developed in this work with the following parameters: mean granulometry of the açai pulp of 0,75 cm; global dry matter of the aqueous mixtures of the pulp of 15%; individual enzymatic concentration of 1% (w:w) on pulp basis; constant orbital agitation of 100 rpm and vigorous agitation by hand every 30 minutes; incubation time and temperature of 4 hours and 50°C, respectively. The enzymatic preparations were tested individually (1x1) in quadruplicate, or combined (2x2; 3x3 and 4x4) in triplicate, totaling 89 tests. The negative control presented a mean yield of extraction of 34,91±11,81%, the lowest of all the tests. The tests performed with the isolated enzymatic preparations have enabled an increase of the extraction yield up to 36,66% compared to the negative control, confirming the important role of the enzymes in the aqueous extraction process. The tests performed with the enzymatic preparations combined 3x3 and 4x4 have increased the extraction yield up to 99,05% compared to the negative control, demonstrating the importance of using the combinations of enzymatic preparations. Among the different tests performed, the enzymatic preparations that stood out are Celluclast 1,5 L, Viscozyme L and Ultrazym AFP by facilitating the oil recovery with a reduce standard deviation (±3,01%). Furthermore, the extraction yield using this combination of three enzymatic preparations is statistically equal to the extraction yield using the four enzymatic preparations, which becomes interesting to the industry due to the cost reduction in the process.Item Acesso aberto (Open Access) Estudo do crescimento bacteriano na presença de óleos essenciais de Dysphania ambrosioides l. e Ocimum campechianum mill. para avaliar seus potenciais como antissépticos bucais(Universidade Federal do Pará, 2015-08-31) MOURA, Luiziana Barbosa; SANTOS, Alberdan Silva; http://lattes.cnpq.br/5976702134131016The aromatic plants like Dysphania ambrosioides (mastruz) and Ocimum campechianum (Alfavaca), that are part of folk medicine in Brazil, have phenylpropanoids and terpenoids compounds in their essential oils as result of secondary metabolism that influence the adaptation and defense of these species in environment; particularly, defense against microrganisms. The objective of this study was to evaluate the antimicrobial activity of essential oils of D. ambrosioides and O. campechianum against pathogenic bacteria mouth. For this they used the disk diffusion method on agar and broth microdilution adapted. The plants were obtained in Santa Izabel do Pará, their leaves were washed and weighed; the essential oil was obtained by hydrodistillation. After, the components of essential oils were identified by gas chromatography-mass spectrometry. Bacteria strains used were: Lactobacillus casei (ATCC 7469), Lactobacillus fermentum (ATCC 9338), Streptococcus mutans (ATCC 25175), Streptococcus oralis (ATCC 10557) and Agregatibacter actinomycetencomitans (ATCC 29522). Bacterial suspensions were prepared for testing and 0.5 McFarland standard. As a positive control we adopted the digluconate of chlorhexidine 0.12%. Inhibition halos were observed for all samples in different concentrations of each essential oil (1%, 5%, 10%, 25%, 50% and 75%). The largest halos were found to A. actinomycetencomitans. In the microdilution test the basil oil inhibited the bacteria S. mutans, the main etiological factor for caries in concentration of 1%; mastruz the oil inhibited L. casei, microorganism that enhances the process of decay in concentrations from 10%; Both oils inhibited the growth of A. actinomycetemcomitans, and can be effective against periodontal disease caused by that pathogen. Plant species of this study produce secondary metabolites classes with potential applications in the development of dental products.Item Acesso aberto (Open Access) Preparação de suportes micro e mesoporosos para imobilização de lipase e aplicação na esterificação do ácido oléico(Universidade Federal do Pará, 2015-09-29) QUARESMA, Francisco Lucio Barbosa; NASCIMENTO, Luis Adriano Santos do; http://lattes.cnpq.br/3720461233595226This study aimed to prepare and characterize heterogeneous catalysts using immobilized enzymes in MCM-41 supports and metakaolin. The supports were functionalized with APTES (an amine) for application to the oleic acid esterification reaction with methanol to produce ester. The characterization of the catalyst was made by XRD, FT-IR, physisorption of N2 and spectrometry of UV. The results show that MCM-41 has been effectively functionalized and immobilized enzymes were, however, there was a partial loss of the hexagonal array of such support, while for metakaolin, both functionalization as immobilization did not occur satisfactorily. Using UV spectrometry, it was found that around 80% of the enzymes were effectively immobilized on MCM-41. This biocatalyst (heterogeneous) was tested in the esterification of oleic acid (molar ratio to oleic acid / methanol was 1:15), at a temperature of 30 °C (100 rpm) for 12h. The obtained conversion was 52% of acid mass to ester, indicating that there viability on the use of MCM-41 amine functionalized for the immobilization of lipase, since catalytic activity of the enzyme was preserved. As for metakaolin, the results showed that, despite having been initially some connection enzymes to metakaolin functionalized, UV spectrometry showed that the anchoring enzyme was not effective in the course of 4 hours, however, new methodological approaches to this material may be applied to ensure the efficient functionalization and immobilization.Item Acesso aberto (Open Access) Produção de ésteres etílicos utilizando rejeito da neutralização do processo do refino dos óleos de buriti, maracujá e castanha-do-pará(Universidade Federal do Pará, 2015-11-27) BRITO, Rogerio da Cunha; NASCIMENTO, Luis Adriano Santos do; http://lattes.cnpq.br/3720461233595226This study deals with ethyl esters obtainment by using three neutralization soapstock generated from the refining process of Buriti oil, Passion fruit and Brazilian nut (Pará nut). At first, acidification of blots was performed to obtain fatty acids concentration, in order to do so it was used a reagent sulfuric acid leaving to react for 50 min at a temperature of around 85 ° C. Moreover, there was formation of three phases: oil component, fat emulsion and phase acidic water, the supernatant phase (oil component) of interest in the possible presence of fatty acids. Characterization by gas chromatography lees of three samples was performed to quantify the fatty acids present therein, the molar masses were determined and also the acid value. The oil components resulting from the acidification reaction were characterized by a saponification number, acid number and the conversion ratios which have the following values: for burity 96.5%, 94.2% for passion fruit and 90.4% and for Brazilian nut. The acidification of the product (oil component) of burity, passion fruit and Brazilian nut were subjected to esterification reaction, in which ethanol was used as a reactant molar ratio 1:30 (oil: ethanol) and 5% catalyst (in relation to the mass of the oil component). The reaction occurred for 60 minutes at a temperature of about 90 ° C. The same parameters were applied for the three reactions. The characterization of ethyl esters was determined by some quality parameters required by ANP, as acidity index, bulk density, kinematic viscosity at 40 ° C and also the content of ethyl esters present in the sample. The rates of conversion of oil components to ethyl esters showed the following percentage: 93.42% (buriti), 91.71% (passion fruit) and 89.38% (Brazilian nut). Despite the higher conversion into ethyl esters have been observed for the Buriti, samples of passion fruit and Brazilian nutt, also had satisfactory results, as those values revealed that the samples have high potential for reuse serving as raw material for production biodiesel.Item Acesso aberto (Open Access) Produção e avaliação da atividade antioxidante de metabólitos secundários de Piper divaricatum G. Meyer sob diferentes condições de cultivo(Universidade Federal do Pará, 2015-08-06) CORPES, Rosana Silva; SILVA, Joyce Kelly do Rosário da; http://lattes.cnpq.br/2278686174214080Many species of the genus Piper are widely distributed in the Amazon and various biological applications because of large structural diversity of its secondary metabolites. The species Piper divaricatum, is endemic in the Amazon and produces in its essential oil high concentrations of methyleugenol (50-90%), an phenylpropanoid with antioxidant and fungicidal properties. Because of its potential applications, the objective of this study was to establish the in vitro cultivation and comparing the biosynthesis of secondary metabolites and antioxidant properties with the in vivo cultivation. For establish the in vitro culture were used shoot apexes on Murashige e Skoog medium with addition of regulator BAP 0.5 mg.mL-1. For in vivo cultivation, micropiles were propagated in the greenhouse in vermiculite and adding nutritious Murashige e Skoog solution. The volatile compounds identified in the leaves of seedlings grown in vivo were methyleugenol, β-elemene and E-β-ocimene, which did not differ from in vivo cultivation, with the exception of 90 days. The in vitro culture of roots was not efficient to produce phenylpropanoids and presented a very different profile compared to the in vivo cultivation of terpenes. In general, for plants in vitro cultivated there was no statistically significant difference in the phenolics compounds content and antioxidant activity in the leaves. However, the antioxidant activity of roots was significant. The results support the hypothesis that in vitro regenerated plants can synthesize metabolites similar the matrix plant and maintain their biological properties.Item Acesso aberto (Open Access) Purificação, caracterização e avaliação de peptídeo antimicrobiano produzido pela cianobactéria amazônica Lyngbya sp. CACIAM 07(Universidade Federal do Pará, 2024-06) PINTO NETO, Joaquim da Silva; SANTOS, Agenor Valadares; http://lattes.cnpq.br/9530734927662735; https://orcid.org/0000-0002-2690-2841Cyanobacteria are photosynthesizing microorganisms that, together with micro and macroalgae, are responsible for most of the oxygen released into the atmosphere. One defense mechanism of cyanobacteria is the production of compounds that exhibit antimicrobial, allelopathic, and antiparasitic activities, protecting the organism against attacks from other species of cyanobacteria, predators, and microorganisms in general. The genus Lyngbya is one of the most studied due to its promising production of molecules of biotechnological interest, such as peptides, proteins, and secondary metabolites, which can be used in various areas to address problems that afflict society and the environment, such as crop pests, food contamination, and the lack of new commercial antimicrobials. The aim of this study was to obtain, evaluate, and characterize the peptide with antimicrobial activity produced by the Amazonian cyanobacterium Lyngbya sp. CACIAM 07. The extract was obtained from the dried biomass of the cyanobacterium, and its antimicrobial activity was evaluated by well diffusion against gram-positive (Bacillus subtilis ATCC 6633, Listeria monocytogenes ATCC 6477, and Corynebacterium fimi NCTC 5) and gram-negative (Salmonella typhimurium ATCC 14021) bacteria. The aqueous extract showed the greatest activity against the target bacteria, inhibiting the growth of three of the four bacteria used in this study: Bacillus subtilis ATCC 6633, Salmonella typhimurium ATCC 14021, and Listeria monocytogenes ATCC 6477. It also inhibited the growth of the cyanobacterium Cyanobium sp. CACIAM 14. The crude aqueous extract was purified using molecular exclusion chromatography and then reversed-phase liquid chromatography. The collected fractions were tested for their antimicrobial and allelopathic activity, with fraction C7B1 showing the best activity against Bacillus subtilis ATCC 6633, Salmonella typhimurium ATCC 14021 and demonstrating allelopathic activity against Synechococcus sp. CACIAM 66. According to the chromatograms, it was possible to determine the size of the molecule, which was 12 kDa, consistent with an antimicrobial peptide. Thus, in this work, it was possible to isolate, analyze, and partially characterize the activity of the possible AMP obtained from Lyngbya sp. CACIAM 07, a cyanobacterium isolated from the Amazon region. We would point out that no molecule with antimicrobial or allelopathic activity from this strain has been described in the literature to date.Item Acesso aberto (Open Access) Vitrificação de tecido ovariano de gata doméstica (Felis catus): um modelo para a preservação da fertilidade em felinos silvestres(Universidade Federal do Pará, 2016-09-06) BRITO, Danielle Cristina Calado de; SANTOS, Regiane Rodrigues dos; http://lattes.cnpq.br/0500967766886604; PIECZARKA, Julio Cesar; http://lattes.cnpq.br/6644368250823351The aim o the present thesis was to develop an efficient vitrification protocol of the ovarian tissue from domestic cat (Felis catus). This study was divided: Phase I: Effect of different basis media during the vitrification of cat ovarian tissue; Phase II: Effect of different sugars (extracellular cryoprotectants) and the vitrification technique for the vitrification of feline ovarian tissue. In phase I, the morphology of preantral follicles was similar (p > 0.05) to fresh control when RPMI-1640 was used as basis medium for vitrification. RPMI-1640 does not contain phenol red, which was found to enhance ethylene glycol (EG) toxicity during vitrification. In phase 2, the percentage of morphologically normal preantral follicles was similar (p > 0.05) to fresh control only when the vitrification medium contained 0.1 or 0.5 M trehalose, instead of sucrose or raffinose at same concentrations. Furthermore, based on parameters such as morphology, cell proliferation and thickness of collagen fibers, it is possibe to assume that efficient vitrification of feline ovarian tissue can be performed by combining trehalose with EG, with or without dimethylsulfoxide (DMSO), applying the solid-surface vitrification (SSV) or ovarian tissue cryosystem (OTC) method. Although vitrification with OTC in the presence of EG did not differ from the other treatments, this protocol presented the highest percentages of preserved preantral follicles (56%), being similar to control (64%). Additionally, no effect on gene regulation was observed after vitrification when apoptosis markers (BAX – protein X associated to Bcl-2), endoplasmic reticulum (ER) stress (ER protein 29 – ERP29), water channels proteins like aquaporins 3 and 9 (AQP3 and AQP9), the membrane ABC transporters ABCB1 and ABCG2, except when the SSV method was applied using only EG as cryoprotectant followed by seven days in vitro culture, where ERP29 up-regulation (ER stress) and AQP9 down-regulation (impaired water transport) were observed. Based on this, it can be concluded that to efficiently preserve feline ovarian tissue, is is necessary the use of a vitrification protocol free of phenol red, supplemented with trehalose, as extracellular cryoprotectant, and EG alone or in combination with DMSO, as intracellular cryoprotectants. Both open (SSV) and closed (OTC) systems are equaly efficient to maintain follicular survival during the vitrification procedure.