Extração, caracterização e atividades biológicas de proteínas da espécie cnidoscolus urens (L.) Arthur

The extraction, chemical and structural characterization of a wide variety of compounds derived from plants has been a major source of bioactive molecules. Several proteases have been isolated in the plant kingdom, with numerous pharmacological and biotechnological applications. Among the proteases isolated from plants, are the fibrinogenolytic, with relevant application in the treatment of disorders in the coagulation cascade, in addition to potential use as a tool in clinical laboratories. In this study, in addition to evaluating the effects of the protein extract of Cnidoscolus urens (L.) Arthur (Euphorbiaceae) in the coagulation cascade also investigates the presence of antimicrobial activity and characterizes the proteolytic activity detected in this extract, aiming to determine their potential pharmacological and biotechnological application. In this way, crude protein extracts obtained from the leaves of C. urens in Tris-HCl 0.05M, NaCl 0.15M, pH 7.5, were precipitated in different concentrations of acetone, and assessed for the presence of proteolytic activity in azocaseína and fibrinogen. The most active fraction (F1.0) in these tests was chosen for assessment of biological activity and biochemical characterization. The Aα chain and Bβ of fibrinogen were completely cleaved at a concentration of 0.18 μg/μL of protein fraction in 4 minutes. Fibrinogenolytic activity presented total inhibition in the presence of E-64 and partial in the presence of EDTA. The fraction demonstrated coagulant activity in plasm and reduced the APTT, demonstrating acting on the factors coagulation of the intrinsic pathway and common, not exerting effects on the PT. Fibrinolytic activity on plasma clot was detected only in SDS-PAGE in high concentrations of fraction, and there were no defibrinating. Although several proteases isolated from plants and venomous animals are classically toxic, the fraction F1.0 of C. urens not expressed hemorrhagic nor hemolytic activities. Fraction F1.0 also showed no antimicrobial activity. In proteolytic activity on the azocasein, the optimal pH was 5.0 and optimum temperature of 60ºC. The enzyme activity has been shown to be sensitive to the presence of salts tested, with inhibition for all compounds. The surfactant triton did not influence the enzyme activity, but the tween-20 and SDS inhibited the activity. In the presence of reducing agents increase in enzyme activity occurred, a typical feature of enzymes belonging to the class of cysteine proteases. Several bands with proteolytic activity were detected in zymogram, in the region of high-molecular-weight, which were inhibited by E-64. In this study, we found that C. urens presents in its constitution cysteine proteases with fibrinogenolytic and procoagulant activity, which may be isolated, with potential application in treatment of bleeding disorders, thrombolytic and clinical laboratory

Caracterização ecofisiológica de sementes de espécies lenhosas da Caatinga

Dormancy is an inherent property of the seeds that define the environmental conditions in which they are able to germinate and their presence is an adaptive trait common in species inhabiting semiarid regions. Moreover, the ability of seedling establishment in these environments has been related to the size, strength and chemical characteristics of the seeds. This study investigated patterns of dormancy and germination speed in tree species of the Caatinga, exploring how the seed size influence the processes of germination, seedling size and biomass allocation. In addition, we aim to investigate the chemical characteristics of the reserves, to verify a possible relationship between nutritional content and the process of seed germination. Therefore, seeds were collected from ten species of woody Caatinga for tests of breaking dormancy, germination and biochemical characterization. Overall, the results show that the scarification treatments mechanical and chemical, and thermal shock influenced the percentage and speed of germination in 50 % of the species, suggesting that they have some level of physical dormancy in the seeds. Biochemical characterization showed the existence of large amounts of carbohydrates in the seeds of all species, low proportion of protein and low amounts of neutral lipids. Using linear regression, we demonstrated the existence of a significant relationship between seed size and the ratio of root/shoot where the largest seeds invested a greater amount of resources for shoot growth. The relationship between germination speed and non-reducing sugar content was also significant, so these compounds is related to the maintenance of physiological seed quality. These results confirm some relationships discussed in the literature for cultivated species, but can be applied to the species native to the Caatinga