Navegando por Palavras-chave "Biorreator"

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    Metagenômica da microbiota envolvida na redução de sulfato e produção de metano em reatores termofílicos de duas fases alimentados com vinhaça
    (Universidade Federal de São Paulo (UNIFESP), 2019-11-11) Silveira, Jessica Teixeira [UNIFESP]; Gregoracci, Gustavo Bueno [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    The present work analysed the specific and functional composition of the microbes involved in vinasse treatment, sulfate removal and methane production, in a thermophilic anaerobic two-phase bioreactor. Biofilm samples were recovered at the end of reactor operation and DNA was extracted with CTAB-chloroform and phenol-chloroform protocols. Sequenced was performed using the shotgun protocol of the Illumina plataform. Data recovery including singlets improved 23,107 hits (7.5%) in R1 and 99,826 hits (6.5%) in R2, with statisticallly significant difference only for the abundance of Thermodesulfovibrio yellowstonii. In R1, a third of the microbiota was composed by T. yellowstonii, which was the main sulfate reducing bacterium (SRB). Still, other SRB were detected in lower abundances. Most SRB use hydrogen, formiate and glucose as substract for sulfate reduction. Ferrodoxins and adenil redutases were detected as sulfate reducing enzymes. Iron reducing species such as Geobacter sulfurreduens and Thermincola potens were also abundant in R1. Fermenting bacteria from Clostridia e Bacilli (Firmicutes) classes were the most abundant in both reactors, and their main fermentation pathway in both reactor was mixed acids fermentation. In R2, butanol biosynthesis and lactate fermentation were also representative pathways. Carbohydrate transformation occurred preferentially through the pentose-phosphate pathway, and through the serine-glyoxate cycle in both reactors. Hydrogenases were the most important funtions connected to respiration in theses reactors as well. Planctomycetaceae family species composed nearly a fifth of R2, suggesting anaerobic ammonium oxidation. Methanogenesis in R2 was performed by Methanobacteriaceae mainly, particularly Methanothermobacter thermautotrophicus. The main methanogenic pathway was performed by hydrogenotrophic archaea, likely due to high concentration of CO2, H2 e formiate. Heterodisulfide reductases were related to methanogenesis. Functional results overall corroborated operational data measured in the reactors.
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    Uso do modo de operação em contínuo para avaliar aspectos fisiológicos de linhagem recombinante de drosophila melanogaster
    (Universidade Federal de São Paulo (UNIFESP), 2019-02-19) Oliveira, Carla Reis [UNIFESP]; Augusto, Elisabeth De Fatima Pires [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Glycoproteins are complex molecules produced preferentially by animal cells, but their quality pattern rely on the knowledge of cell metabolism and requires engineering tools to ensure greater uniformity and productivity. As long as reports on the metabolism of insect cells are scarce, researches on the physiological aspects of Drosophila*melanogaster are of utmost importance to promote their usage. In order to contribute on the knowledge of this metabolism, the present work aimed to identify growth limiting and inhibitory factors of S2AcRVGP2, a D.*melanogaster S2 cell line stably transfected for the synthesis of Rabies virus glycoprotein (GPV). This study was conducted using TC100 based medium in spinner flasks and in benchtop bioreactor stirred at 100 rpm and with bubbleLfree aeration system. Exploratory cultures on batch mode were carried out with different concentrations of LLcystine (CYS2) and magnesium (Mg2+). The influence on the metabolism of different glucose concentrations (GLC) in the feed medium was studied in a continuous mode. It has been shown that lactate and ammonium did not seem to be growth inhibiting. The supplementation with CYS2 11 mg/L enhanced GPV expression by 75% in batch with initial supplementation and 21% in batch with pulse addition at the end of exponential phase. The evaluation of Mg2+ influence on cell growth kinetic parameters showed that when cultivated with half of its basal concentration, cell production increased 37%, but when cultured on [Mg2+] inferior to 121 mg/L, it limited the growth rate and altered cell membrane morphology. Continuous culture with glucose concentration above 3.75 g/L in the feed medium indicated overflow on glycolysis with consequently limitation of cell growth. The metabolism shifted to a more balanced state when fed with glucose concentration under 2.5 g/L. Cell production increased by 40% and GPV expression doubled in the lowest glucose concentration evaluated (1.25 g/L).