Covalent flavinylation of L-aspartate oxidase from Escherichia coli using N6-(6-carboxyhexyl)-FAD succinimidoester.

L-Aspartate oxidase is a flavoprotein catalyzing the first step in the de novo biosynthesis of pyridine nucleotides in E. coli. Binding of FAD to L-aspartate oxidase is relatively weak (K(d) 6.7 x 10(-7) M), resulting in partial loss of the coenzyme under many experimental conditions. Only the three-dimensional structure of the apo-enzyme has been obtained so far. In order to probe the flavin-binding site of the enzyme, apo-L-aspartate oxidase has been reacted with N6-(6-carboxyhexyl)-FAD succinimidoester. The structural characterization of the resulting N6-(6-carbamoylxyhexyl)FAD-L-aspartate oxidase shows the covalent incorporation of 1 FAD-analog/monomer. Residue Lys38 was identified as the target of the covalent modification. N6-(6-carbamoylxyhexyl)-FAD-L-aspartate oxidase shows only 2% catalytic activity as compared to the native enzyme. Comparison of some properties of the flavinylated and native enzymes suggests that, although the flavin is covalently bound to the former in the region predicted from molecular modeling studies, the microenvironment around the isoallossazine is different in the two forms

Covalent flavinylation of L-aspartate oxidase from Escherichia coli using N6-(6-carboxyhexyl)-FAD succinidoester

L-Aspartate oxidase is a flavoprotein catalyzing the first step in the de novo biosynthesis of pyridine nucleotides in E. coli. Binding of FAD to L- aspartate oxidase is relatively weak (K(d) 6.7 x 10-7 M), resulting in partial loss of the coenzyme under many experimental conditions. Only the three-dimensional structure of the apo-enzyme has been obtained so far. In order to probe the flavin-binding site of the enzyme, apo-L-aspartate oxidase has been reacted with N6-(6-carboxyhexyl)FAD succinimidoester. The structural characterization of the resulting N6-(6-carbamoylxyhexyl)FAD-L- aspartate oxidase shows the covalent incorporation of 1 FAD-analog/monomer. Residue Lys38 was identified as the target of the covalent modification. N6- (6-carbamoylxyhexyl)-FAD-L-aspartate oxidase shows only 2% catalytic activity as compared to the native enzyme. Comparison of some properties of the flavinylated and native enzymes suggests that, although the flavin is covalently bound to the former in the region predicted from molecular modeling studies, the microenvironment around the isoallossazine is different in the two forms

Surfactantes reativos não-iônicos em polimerização em emulsão de látices de acetato de vinila - vinil neodecanoato: influência nas propriedades de barreira à água Nonionic reactive surfactants in emulsion polymerization of vinyl acetate - vinyl neodecanoate latexes: influence on the water barrier properties

A indústria de tintas é grande consumidora de látex obtido por polimerização em emulsão. Os surfactantes, essenciais à estabilidade do látex, exercem papel fundamental na produção e na aplicação destes polímeros. Contudo, podem também produzir efeitos adversos nas propriedades do produto, em razão de sua adsorção física às partículas de polímero. Os surfactantes não ligados podem migrar através do filme para as interfaces, formando agregados que podem aumentar a sensibilidade à água, afetando desta forma as propriedades de barreira. Um caminho promissor para minimizar este efeito dos surfactantes convencionais tem sido o uso de surfactantes polimerizáveis, ou reativos, que estão covalentemente ligados ao polímero e, desta forma, não podem ser dessorvidos e migrarem durante a formação do filme. Neste trabalho foram preparados látices de acetato de vinila - vinil neodecanoato (VeoVa 10®), estabilizados com surfactantes não-iônicos convencionais e reativos, e avaliado o desempenho dos filmes obtidos a partir destes látices. Os resultados demonstraram que o uso de surfactantes não-iônicos polimerizáveis pode, sob determinadas condições, trazer ganhos para as propriedades de barreira.<br>The paint industry is a huge consumer of latex from emulsion polymerization. The surfactants, essential to the stability of the latex, play a crucial role in the production and application of emulsion polymers. However, they can also have adverse effects on product properties due to their physical adsorption on the polymer particles. The unbound surfactants can migrate through the film toward the interfaces forming aggregates which increase water sensitivity of the film, thus affecting its barrier properties. A promising way to reduce the negative effects of the conventional surfactants is to use polymerizable or reactive surfactants (surfmers) that are covalently linked to the polymer, which avoids its desorption and migration during the film formation. In this work vinyl acetate - vinyl neodecanoate (VeoVa 10®) latexes, stabilized with conventional and reactive nonionic surfactant, were prepared and the performance of these films was evaluated. It was noted that latexes stabilized with nonionic polymerizable surfactants can bring, under certain conditions, better barrier properties