Fibrinolytic proteases are enzymes that degrade fibrin; these enzymes are a promising alternative for thrombolytic therapy, and microorganisms produce them. The aim of this study was to evaluate the optimum conditions for the integrated production and purification of fibrinolytic protease from Bacillus sp. UFPEDA 485. Extractive fermentation was carried out in a culture medium containing soybean flour and by adding polyethylene glycol (PEG) and Na2SO4 according to a 2(3) experimental design. In all assays, the enzyme preferentially partitioned to the bottom phase (K < 1), with an optimum activity of 835 U ml(-1) in the bottom phase (salt-rich phase). The best conditions for extractive fermentation were obtained with 18 % PEG 8000 and 13 % Na2SO4. Characterization showed that it is a metalloprotease, as a strong inhibition-residual activity of 3.13 %-occurred in the presence of ethylenediaminetetraacetic acid. It was also observed that enzymatic activity was stimulated in the presence of ions: CaCl2 (440 %), MgCl2 (440 %), FeSO4 (268 %), and KCl (268 %). The obtained results indicate that the use of a low-cost substrate and the integration of fermentation with an aqueous two-phase system extraction may be an interesting alternative for the production of fibrinolytic protease.

-The objective of this work was to investigate, on a flask scale, the production of bioemulsifiers by Yarrowia lipolytica in the presence of sea water, supplemented with nitrogen and phosphate sources, using diesel oil as substrate. A full 2 4 factorial design was conducted to investigate the effects and interactions of the nutrient concentrations (diesel oil, urea, ammonium sulfate and monobasic potassium phosphate) on the response variables: emulsification activity and surface tension of the cell-free cultures. High emulsification activities (> 5,4 UEA) were determined after 168 h in all the experiments. The interactions among diesel oil, urea and monobasic potassium phosphate favored the emulsification with statistical significance. A correlation between the increase of emulsification activity and the reduction of surface tension was not identified.

The capacity of fibrinolytic enzymes to degrade blood clots makes them of high relevance in medicine and in the pharmaceutical industry. In this work, forty-three microorganisms of the genus Bacillus were evaluated for their potential to produce fibrinolytic proteases. Thirty bacteria were confirmed as producers of fibrinolytic enzymes, the best results obtained for the strain Bacillus amyloliquefaciens UFPEDA 485. The optimization of the enzyme production conditions was done by a central composite design (CCD) star 2 3 that allowed to define the optimal conditions for soybean flour and glucose concentrations and agitation rate. The highest fibrinolytic activity (FA) of 813 U mL -1 and a degradation of blood clot in vitro of 62% were obtained in a medium with 2% (w/v) of soybean flour and 1% (w/v) glucose at 200 rpm after 48 h of cultivation, at pH 7.2 and 37°C. The obtained fibrinolytic enzyme was characterized biochemically. Fibrinolytic activity was inhibited by PMSF (fluoride methylphenylsulfonyl -C 7 H 7 FO 2 S) 91.52% and EDTA (ethylenediaminetetraacetic acid -C 10 H 16 N 2 O 8 ) 89.4%, confirming to be a serine-metallo protease. The optimum pH and temperature were 7.0 and 37 o C, respectively, and the enzyme was stable for 12 h. The fibrinolytic activity at physiological conditions of this enzyme produced by Bacillus amyloliquefaciens UFPEDA 485, as well as its long term stability, demonstrate that it has suitable characteristics for human and veterinary applications, and promises to be a powerful drug for the treatment of vascular diseases.

RESUMO -Microalgae are potential sources of high value chemicals and bioactive compounds. To promote the cell productivity, culture conditions such as carbon and nitrogen sources, irradiance, temperature, bioreactor design have been investigated. Under mixotrophic conditions, some microalgae grows rapidly and to have a higher growth rate when compared to photoautotrophic conditions. This works reports Chlorella vulgaris production under mixotrophic conditions using glycerol and corn steep licor. The factorial design (2 2 plus star central) was performed with 0.5-1.5% gycerol (C gly ) and 0.5-1.5% corn steep liquor (C csl ) using maximum cell concentration (X m ), cell productivity (P X ) as response. Results from the response surface methodology were that the X m of 1.52 g L -1 was obtained using 0.9 % of G and 1.2 % of CSL. The highest P X value was 232 mg L -1 day -1 with 1.5% of G and CSL. Therefore, these models can be useful to predict maximum cell concentration in mixotrophic cultures.

Alternative and practical classes are effective in assisting teachers and students in the teaching-learning processes, aiming at good examples in the construction of knowledge, serving as a complement to traditional classes, which in most cases become exhausting due to lack of investment and innovation. The respiratory system is of great importance for human life as it is responsible for the absorption of oxygen through the airways and lungs. Observing the difficulties faced by teachers in the classroom due to limited resources within the school, this work had the objective of implementing an alternative methodology to improve the acquisition of knowledge about biophysics of the respiratory system. The research was carried out with a group of the 3rd year of High School of the Aluísio Germano School in the city of Carpina - PE. Two identical questionnaires were applied: one after the traditional class and another after the alternative class with the application of the Quiz game. The results indicated that there was a considerable increase in the correctness of the questions in the second questionnaire compared to the first one. In addition, it was observed that, during the class that used the game, the students showed more interest in knowing and learning about the subject, having an increase of interaction and totaling 85% the level of interaction in the proposed activities, which corroborates with other authors in the literature. In this way, it was noticeable that the use of playful resources associated with the practices can be great allies in the teaching-learning process.

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