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Navegando Região Metropolitana de Goiânia (RMG) por Por Orientador "Abdelnur, Patrícia Verardi"
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Item Metabolômica de leveduras fermentadoras de xilose na identificação de alvos metabólicos para aumento da produção de etanol 2g(Universidade Federal de Goiás, 2019-02-18) Cavalcanti, Christiane Gonçalves Campos; Rodrigue, Clenilson Martins; http://lattes.cnpq.br/9689635455677053; Abdelnur, Patrícia Verardi; http://lattes.cnpq.br/0386805742968055; Abdelnur, Patrícia Verardi; Vaz, Boniek Gontijo; Côrrea, Mauro Vicentini; Parachin, Nádia Skorupa; Silva, Caio de Oliveira GorgulhoOne of the challenges in the production of second generation (2G) ethanol is in the development of efficient yeasts to convert xylose into ethanol in the process of fermenting sugars. Thus, the comprehensive analysis of xylose fermenting yeast metabolism is essential to facilitate the identification of the limiting factors in the conversion metabolism of this pentose and, thus, help in the construction of more efficient genetically modified strains. The objective of this work was to use an advanced analytical tool, the metabolomics, to quantify the main metabolites related to the xylose to ethanol conversion pathways in four species of yeast xyloses: Scheffersomyces stipitis, Spathaspora passalidarum, Spathaspora arborariae and Candida Tenuis. The yeasts were grown in xylose under two different growth conditions, aerobic and microaerobic. The preparation of the sample for metabolic analysis included the steps of quenching and extraction of the metabolites, for which the protocols of ice cold methanol and boiling ethanol, were used, respectively. The quantification of the metabolites was performed by ultra high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/ MS), using ion-pair chromatography and hydrophilic interaction liquid chromatography. The data obtained in the quantification were processed and treated through analysis of variance (ANOVA) using RStudio software. A total of eleven metabolites of the xylose metabolism pathway were quantified and validated through metabolic flow analysis (MFA) with R2>90. These metabolites were used to construct the metabolic flow of the three species of yeasts (Scheffersomyces stipitis, Spathaspora passalidarum and Spathaspora arborariae) analyzed. The comparison of molecular targets in yeast xylose showed that the S. stipitis and S. passalidarum species have the best yields of ethanol when grown with limited oxygen. The metabolomic analysis performed in this study may help in the genetic improvement of these microorganisms and in the production of 2G ethanol.Item Metabolômica de macrofungos cultivados em torta do caroço de algodão por espectrometria de massas(Universidade Federal de Goiás, 2020-02-19) Souza, Miria de Almeida; Abdelnur, Patrícia Verardi; http://lattes.cnpq.br/0386805742968055; Abdelnur, Patrícia Verardi; Silva, Caio de Oliveira Gorgulho; Kato, LucíliaThe large production of agro-industrial waste arouses the search for sustainable alternatives. These residues are a great source of energy and biomass, and can be used for energy generation, biofuels and chemicals. Cotton is a crop that has great commercial importance. In addition to the production of fibers used in the manufacture of textiles, the seed is used to produce oil (liquid fraction) and cake/bran (solid fraction). Cotton seed cake/bran (TCA) has a significant amount of protein and carbohydrates, which can be used for animal nutrition. However, TCA has some antinutritional or toxic compounds for animal nutrition, such as gossypol. The use of microorganisms, among them, macrofungi has been a strategy to degrade/detoxify such chemical compounds and thus facilitate their use in mono or polygastric nutrition. However, it is necessary to identify the bioactive chemical compounds produced by the macrofungi, and in this case, the metabolomics based on mass spectrometry can be an applied tool to characterize the low molecular weight fungal metabolites. Thus, this work characterized the metabolites present in cottonseed cake after solid state fermentation of macrofungi, using untargeted metabolomics by Ultra-High Performance Liquid Chromatography - Mass Spectrometry (UHPLC-ESI-MS / MS). The resulting data were submitted to Principal Component Analysis (PCA) and compared to the database of Chemical Entities of Biological Interest and Kyoto Encyclopedia of Genes and Genomes. The PCAs of the macrofungi Schizophyllum commune (FPB 117) and Fistulina hepatica (CC102) showed significant changes in the profile of chemical compounds in cottonseed cake (in natura) after fungal fermentation. In the fermented (TCA + macrofungus) it was possible to identify chemical compounds of the classes of lipids, amino acids, oligosaccharides, nucleosides and nucleotides. The fermented with Pleurotus pulmonarius EF88 showed a chromatographic profile very close to the control, indicating the microbial cultivation process under the tested conditions did not show significant growth of the macrofungus. The untargeted metabolomics tool applied in this work proved to be efficient for the identification and characterization of metabolites from microbial fermentative processes in plant biomass. Such information contributes to studies involving the deconstruction of lignocellulosic biomass and the animal nutrition industry.Item Desenvolvimento de métodos baseados em espectrometria de massas e cromatografia líquida para análise de compostos químicos produzidos por bioconversão de glicerina(Universidade Federal de Goiás, 2018-07-19) Vieira, Flávia Soares; Rodrigues, Clenilson Martins; http://lattes.cnpq.br/9689635455677053; Abdelnur, Patrícia Verardi; http://lattes.cnpq.br/0386805742968055; Abdelnur, Patrícia Verardi; Rodrigues, Clenilson Martins; Kato, Lucília; Correia, Mauro VicentiniBiodiesel production has been increasing every year, which has generated surpluses of glycerin, the main co-product of the transesterification reaction. In the search for applications to this surplus, microbial fermentation has been highlighted as an alternative to add value to this co- product. In this process, microorganisms are used to convert this abundant carbon source into compounds of high economic value, including organic acids and polyols. However, there are still some technological challenges, such as low yield during the fermentation processes and high dependence of new analytical methods, more sensitive and selective, able to identify and quantify the compounds produced. The objective of this work was to establish new analytical methodologies to identify and quantify compounds from bioconversion processes involving the production of organic acids and polyols. To guarantee the quantification of the target compounds, methods based on ultra-high- performance liquid chromatography (UHPLC) were developed and the photodiode array (PDA) and mass spectrometry (MS) detectors were evaluated as alternatives for the monitoring of these two classes of chemical compounds. The analytes investigated in this study were short chain organic acids: citric acid, fumaric acid, glyceric acid, kojic acid, lactic acid, muconic acid, oxalic acid, propionic acid, succinic acid, xylonic acid; and polyols with spatial isomers, including polyols with 4 carbons (C4: erythritol and threitol), 5 carbons (C5: arabitol, ribitol, xylitol), and 6 carbons (C6: dulcitol, iditol, mannitol, sorbitol) and 7 carbons (C7: volemitol). The UHPLC-PDA method developed for the analysis of organic acids was validated by parameters of linearity, limit of detection and quantification, precision and accuracy. The analytical curves showed good linearity with determination coefficients (R²) higher than 0.999 for all organic acids. Precision and accuracy were adequate with intra-day and inter-day variation coefficients below 4.06% and recovery ranged from 94.90% to 109.63%. The developed UHPLC- MS methods presented greater sensitivity and selectivity in the detection of organic acids and polyols, being able to monitor these analytes even in low quantity. The analytical methods developed in this work are crucial tools in the selection or screening of promising microorganisms for low added value substrates conversion and important to optimize biotechnological processes for production of these compounds of interest.