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Estudos funcionais de inibidores de cisteíno peptidases da cana-de-açúcar e caracterização de uma cisteíno peptidase de Sphenophorus levis, uma importante praga da cultura canavieira
(Universidade Federal de São Carlos, 2009-04-30) Dellamano, Marcia; Silva, Flávio Henrique da; https://lattes.cnpq.br/1757309852446263; https://lattes.cnpq.br/6172605531468633
Cystatins are proteins that inhibit specifically cysteine peptidases. The Canecystatin 1 gene which codifies a protein containing 106 amino acids residues was identified in sugarcane and possesses significant similarity with Oryzacystatin, a cystatin from rice. In order to obtain a cystatin with improved activity, direct evolution experiments were carried out. A DNA shuffling library was constructed using these two cystatins. One clone named A10PL3 obtained from these shuffled cystatins was selected, expressed in E.coli, purified and an analyzed by activity assays. These results showed that the activity of hybrid protein A10PL3 increased, in particular regarding its inhibitory activity on cathepsin B compared with its two precursors. The present study aimed to revert the changes of individual clone A10PL3 through site-directed mutations, generating three mutant cystatins: Mutant I (Thr17Ile), Mutant II (Gln 84Leu) and Mutant III (Thr17Ile); (Gln84Leu). Assays for inhibitory activity against the human cathepsins B and L were performed. Structural studies were also made by means of molecular modeling of proteins by homology that were enabled to understand the molecular mechanisms related to improvement of the inhibitory activity of these cystatins. These studies therefore corroborate with the observed data previously which demonstrated the improvement of specific protein A10PL3 in cathepsin B inhibition (Ki 16 nM) in relation to their parents. The mutants I, II and III, did not present improvement in inhibitory activity against cathepsin B. The structural studies revealed that the mutations performed on cystatin A10PL3 destabilized the hydrophobic core making it more flexible, thus increasing the inhibitory activity on cathepsin B. The absence of interactions underlying the hydrophobic core resulted in a trend of lower solubility, probably due to their inability to adopt a compact formation, which resulted in the exposure of some residues which are part of that core, which can lead to aggregation and also contribute to increasing the flexibility of cystatin, influencing their inhibitory activity.
Aplicação de nanopartículas em filmes utilizados em embalagens para alimentos
(Universidade Federal de São Carlos, 2009-04-30) Aouada, Márcia Regina de Moura; Mattoso, Luiz Henrique Capparelli; https://lattes.cnpq.br/5839043594908917; https://lattes.cnpq.br/2870655742911951
Dispersions composed of chitosan (CS) were obtained by the template polymerization (CS-PMAA) and ionotropic gelatinization (CS-TPP). The nanoparticles were analyzed by FT-IR analysis, particle size, zeta potential, electron microscopy (transmission and scanning), and TGA analysis. The nanoparticles obtained presented a very homogeneous morphology showing a quite uniform particles size distribution and a rather spherical shape. The particle size and zeta potential were dependent on the chitosan concentration used during the preparation method and pH. Different particles were incorporated in the hydroxypropyl methylcellulose (HPMC) films in order to improve their properties. For this, a 3% HPMC (in w:v %) solution was prepared and added in this nanoparticles solutions of both syntheses. The films were characterized by mechanical (tensile, elongation and elastic modulus), barrier [water vapor (WVP) and oxygen], thickness, morphological, and thermal properties. The WVP of the control HPMC film was 0.79 g mm/kPa h m2. The WVP decreased significantly when CS-PMAA nanoparticles were incorporated to HPMC films. For example, WVP decreased to 0.64; 0.59 and 0.47 g mm/kPa h m2 for films containing 59, 82 and 110 nm, respectively. For CS-TPP nanoparticles the WVP decreased to 0.58; 0.45 and 0.33 g mm/kPa h m2 for films containing nanoparticles with 220, 110 and 85 nm, respectively. The same way, the mechanical properties of the HPMC films were strongly improved when CS nanoparticles were incorporated. In this way, HPMC films containing nanoparticles could be considered as promising materials for application in packaging for preservation of foods.
Obtenção e caracterização de perovskitas supercondutoras monocristalinas do tipo YxPr1-xBa2Cu3O7-δ dopadas por Fe, Ni, Ga e Zn
(Universidade Federal de São Carlos, 2009-04-30) Cichetto Júnior, Leonélio; Moreira, Fernando Manuel Araújo; https://lattes.cnpq.br/1809254923092721; https://lattes.cnpq.br/7434408308350513
The Praseodymium is known to have a singular behavior in the family of cuprate superconductors of high critical temperature of the type RE-123. When these cuprates are doped with Praseodymium apparently critical temperature decreases, with a metalinsulator transition. The origin of this behavior is of fundamental interest, and it may provide an explanation for the mechanism of superconductivity at high temperatures, since any reasonable theory for this mechanism must explain the behavior of the system Y1-xPrxBa2Cu3O7-δ, also called YPr-123. Through the experience gained by the research group to which we belong, and where this research project was developed, we continue the work begun by the PhD students Cecilia Stari and Alexandre Lanfredi. These two students have developed studies of polycrystalline YPr-123 family, and single crystals of YBa2Cu3O7-δ (Y-123) obtained by the method of Sol-Gel grown in crucibles of SnO2, respectively. None of them worked with the collection and characterization of single-crystalline samples of the system YPr-123, with special emphasis on Pr-rich samples, and that pure one, or PrBa2Cu3O7-δ (Pr-123). Studying to obtain and characterize samples of single-crystalline superconducting Pr-123 is the main goal of our Project Master. Besides obtaining samples of single-crystalline YPr-123 samples, in this master's work was studied the influence of cation doping in single crystals of composition Y1-xPrxBa2Cu3O7-δ (YPr123) for 0 ≤ x ≤ 1. Doping were performed with the elements Ga, Zn, Fe and Ni to study the behavior of superconductivity in these types of crystals. To obtain samples with a high degree of purity necessary for the study of their basic properties, were made our own SnO2 crucibles with high resistance to corrosion caused by self-flow used in the growth of crystals. Were also used Al2O3 and ZrO2CeO crucibles. The chemical composition and structural quality of these samples were analyzed by x-ray diffraction (XRD) and scanning electron microscopy (SEM) associated with the techniques of micro-analysis by energy dispersive X-ray (EDX). Also, we have used measures of the magnetic susceptibility for the characterization of superconducting properties of all samples. The results indicated that the whole YPr-123 family shows superconductivity, which is still very controversial in the literature. Specifically about the compound PrBa2Cu3O7-δ we verified that the sample is a superconductor, with critical temperature around 98K. This important result allows us to conclude that the relevant factor to observe the presence of superconductivity in this family is the Argon atmosphere during the growth of single crystal, as has been pointed out by previous publications of our research group. The results found here characterize this work as unique in the study of single crystal samples of the family YPr-123.