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Estudo de condições operacionais na produção de penicilina G acilase por diferentes microrganismos
(Universidade Federal de São Carlos, 2006-12-20) Oguri, Renata Tieko; Giordano, Raquel de Lima Camargo; https://lattes.cnpq.br/9847634405993955
Penicillin G acylase (PGA) is an enzyme of major impact in human health for catalyzing the deacilation of the penicillin G, to yield 6-aminopenicillanic acid (6-APA), key intermediate in the semi-synthetic antibiotics production. Several microorganisms produce PGA, but Bacillus megaterium is capable of secreting it to the medium. PGA from B. megaterium has been studied in DEQ-UFSCar. Xanthomonas campestris presents in your DNA the genetic sequence that codify acylase penicillin and tests with a strain of this microorganism from Departamento de Genética e Evolução da UFSCar showed PGA extracellular production. Studies in PGA production were initiated with the microorganism stored in DEQ-UFSCar. At the same time, strains of X. campestris from Coleção de Culturas Tropical (CCT) and Instituto Agronômico do Paraná (IAPAR) were tested. However, assays with these strains showed little cellular growth and no enzymatic activity. The microorganism reactivated was send to Fundação André Toselo for identification. The result showed that the microorganism was Bacillus megaterium. Therefore, at the same time that realized adaptations tries with the new strain of X. campestris, studies of production operational conditions of PGA were performed with strain of Bacillus megaterium, in shaking flasks and bioreactor, as well the enzyme characterization Germination/ propagation of B. megaterium experiments, at 30°C, carried out in shaking flasks showed that the growth time of microorganism in the inoculum step was 24 hours and maximum specific growth rate was µmáx = 0.33h-1. Assays in shaking flasks were carried out in shaker at 30ºC and 300 rpm intend to determine the most adequate pH for the enzyme production in both, microorganism propagation phase and enzyme production phase The results showed that the best pHs among the tested ones were pH 8.0 and 7.0, for the propagation and production phases, respectively, with 355 IU/L. The culture medium with preferentially consumed amino acids 20,0 g/L, potassium phenylacetate 3,5 g/L, solution with differents salts 0,22 g/L and cheese whey 20,0 g/L was the more efficient in PGA production, carried out maximum cellular growth and enzymatic activity, 4,59 g/L and 592UI/L, respectively, in 36 hours. Assays in a 2-L bioreactor, with production medium in the presence of amino acids preferably consumed 10.0 g/L, cheese whey 20.0 g/L, solution of salts 0.22 g/L and inducer potassium phenylacetate 3.5 g/L, indicated that the addition of nutrients during the fermentation is a efficient strategy, providing enzyme production increase. In the study of the influence of oxygen dissolved, carried out in bioreactor too, maximum enzymatic activity was 451 IU/L with 10% of saturation. Moreover, the maximum enzyme production was observed in the bioreactor and not in shaking flasks, like in all experiments until this. In the stage of characterization of the penicillin G acylase from Bacillus megaterium, optimum values of temperature and pH were, 47°C and 8.0, respectively. The Michaelis Menten model fitted resulting in estimated Km and Vmáx parameters values of 1.51 mM and 1.1*10-3 mmol/min*IU, respectively. The activation energy estimated was 27.12 KJ/mol.The results showed that PGA from Bacillus megaterium deactivate completely after 45 minutes of incubation at 60°C and 90 minutes in pH 11.0.
Estudo de oportunidades de reúso de águas residuárias em campi universitários - estudo de caso da Universidade Federal de São Carlos
(Universidade Federal de São Carlos, 2006-12-20) Antoniosi, Patricia Maria Pedro Bom; Salvador, Nemésio Neves Batista; https://lattes.cnpq.br/9930582531600560;
Water, one of the most important natural resources necessary to the man’s life, is in the planet distributed irregularly and committed by its bad use. Several sections look for solutions for the subjects of the shortage of the resources, But the population don t notice its importance inside of the chain consumption and generation of residues, it still intensifies more the problem. For its appropriate administration, seeking the sustainability and the development, conferences on environment brought the surface many ways to reached those objects, explaining concepts of consumption reduction, reuse and recycling. This work aims to integrate those concepts inside of the systems of water supply how principal objective the reuse of the water resources, The wastewaters are a alternative source for the supplying in human activities. The Universidade Federal de São Carlos was chosen, for having uses of many natures and production of the more differentiated types of waters reuse. Through listings, characterizing the uses of the water, they were lifted up the generating activities of the possible water for the reuse and the consuming activities to the campus already existent in São Carlos, identifying the possible opportunities of reuse or recycle the wastewaters inside of the university. Seen the great potentiality of applying wastewaters in some activities in universities, the new project of the campus of Sorocaba, segregated systems of sewer and It was proposed a reservoir of water supply with double camera for drinking water and reuse water, in same structure and supplying the whole university, improving a economy of the water and electric power resource.