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URI permanente desta comunidadehttps://thoth.dti.ufv.br/handle/123456789/3352
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Item Functional complementation of a yeast knockout strain by Schistosoma mansoni Rho1 GTPase in the presence of caffeine, an agent that affects mutants defective in the protein kinase C signal transduction pathway(Instituto Oswaldo Cruz, Ministério da Saúde, 2006-05-26) Aguiar, Pedro H. N. de; Santos, Débora N.; Lobo, Francisco P.; Santos, Túlio M.; Macedo, Andréa M.; Pena, Sérgio D. J.; Machado, Carlos R.; Franco, Glória R.In a previous study, the Schistosoma mansoni Rho1 protein was able to complement Rho1 null mutant Saccharomyces cerevisiae cells at restrictive temperatures and under osmotic stress (low calcium concentration) better than the human homologue (RhoA). It is known that under osmotic stress, the S. cerevisiae Rho1 triggers two distinct pathways: activation of the membrane 1,3-beta-glucan synthase enzymatic complex and activation of the protein kinase C1 signal transduction pathway, promoting the transcription of response genes. In the present work the SmRho1 protein and its mutants smrho1E97P, smrho1L101T, and smrho1E97P, L101T were used to try to clarify the basis for the differential complementation of Rho1 knockout yeast strain by the human and S. mansoni genes. Experiments of functional complementation in the presence of caffeine and in the presence of the osmotic regulator sorbitol were conducted. SmRho1 and its mutants showed a differential complementation of the yeast cells in the presence of caffeine, since smrho1E97P and smrho1E97P, L101T mutants showed a delay in the growth when compared to the yeast complemented with the wild type SmRho1. However, in the presence of sorbitol and caffeine the wild type SmRho1 and mutants showed a similar complementation phenotype, as they allowed yeast growth in all caffeine concentrations tested.Item Coffee protein profiles during fermentation using different yeast inoculation methods(Empresa Brasileira de Pesquisa Agropecuária - Embrapa, 2020) Bressani, Ana Paula Pereira; Martinez, Silvia Juliana; Vilela, Leonardo de Figueiredo; Dias, Disney Ribeiro; Schwan, Rosane FreitasThe objective of this work was to evaluate the protein profiles of natural and semidry fermented Coffea arabica, either subjected to treatments with different yeast inoculation methods with starter culture or to na uninoculated control. Saccharomyces cerevisiae CCMA 0543 and Candida parapsilosis CCMA 0544 were separately inoculated into coffee by directly spraying the cherries on a terrace or in buckets, for 16 hours before sun drying. Protein quantification showed a significant difference between the protein profiles of the samples collected after natural dry fermentation. The MALDI-TOF MS analysis generated a list of 96 peaks with different massto- charge ratios (m/z) in the samples collected at the beginning and the end of fermentation. The highest number of peaks in the natural dry coffee was observed at the end of fermentation in the samples inoculated with S. cerevisiae CCMA 0543, in bucket, and in C. parapsilosis CCMA 0544 sprayed on the terrace. However, in the semidry processed coffee, the highest number of peaks was observed in the initial fermentation, with a decrease in the peptide peaks after fermentation. The fermentation with different microorganisms, processing types, and inoculation methods affects m/z profiles, influencing the types of proteins found in coffee.