Structural and Biochemical Characterization of Peroxiredoxin Q beta from Xylella fastidiosa CATALYTIC MECHANISM and HIGH REACTIVITY

The phytopathogenic bacterium Xylella fastidiosa is the etiological agent of various plant diseases. To survive under oxidative stress imposed by the host, microorganisms express antioxidant proteins, including cysteine-based peroxidases named peroxiredoxins. This work is a comprehensive analysis of the catalysis performed by PrxQ from X. fastidiosa (XfPrxQ) that belongs to a peroxiredoxin class still poorly characterized and previously considered as moderately reactive toward hydroperoxides. Contrary to these assumptions, our competitive kinetics studies have shown that the second-order rate constants of the peroxidase reactions of XfPrxQ with hydrogen peroxide and peroxynitrite are in the order of 107 and 106 M(-1) s(-1), respectively, which are as fast as the most efficient peroxidases. The XfPrxQ disulfides were only slightly reducible by dithiothreitol; therefore, the identification of a thioredoxin system as the probable biological reductant of XfPrxQ was a relevant finding. We also showed by site-specific mutagenesis and mass spectrometry that an intramolecular disulfide bond between Cys-47 and Cys-83 is generated during the catalytic cycle. Furthermore, we elucidated the crystal structure of XfPrxQ C47S in which Ser-47 and Cys-83 lie similar to 12.3 angstrom apart. Therefore, significant conformational changes are required for disulfide bond formation. In fact, circular dichroism data indicated that there was a significant redox-dependent unfolding of alpha-helices, which is probably triggered by the peroxidatic cysteine oxidation. Finally, we proposed a model that takes data from this work as well data as from the literature into account.

Influence of levels and forms of selenium associated with levels of vitamins C and E on the performance, yield and composition of tilapia fillet

Objetivou-se avaliar a influência da vitamina C (VC), vitamina E (VE) e de duas formas de selênio sobre o desempenho, rendimento e qualidade do filé de tilápia Nilótica. O experimento foi realizado num delineamento inteiramente casualizado composto por seis tratamentos, os quais foram constituídos pela adição de 100, 200 e 400 mg kg-1 das vitaminas C e E, associados com 0,1, 0,2 e 0,4 mg kg-1 de duas formas de selênio. Cada tratamento foi constituído por cinco repetições de 30 peixes cada. A dieta com 200 mg kg-1 de VC e VE + 0,2 mg kg-1 de selênio orgânico proporcionou ganho de peso, comprimento corporal e fator de conversão alimentar semelhantes aos tratamentos com 400 mg kg-1 de VC e VE + 0,4 mg kg-1 de selênio orgânico ou inorgânico. A adição de selênio orgânico à dieta melhorou o ganho de peso e o fator de conversão alimentar em comparação com o selênio inorgânico. A dieta com 0,2 mg kg-1 de selênio orgânico apresentou nível de glutationa peroxidase igual ao da dieta com 0,4 mg kg-1 de selênio inorgânico. Os rendimentos de carcaça e filé não foram influenciados pelos tratamentos, entretanto houve efeito dos tratamentos na composição química do filé.