Physico-chemical studies of molecular interactions between non-ionic surfactants and bovine serum albumin.

Surfactants, particularly non-ionic types, are often added to prevent and/or minimize protein aggregation during fermentation, purification, freeze-drying, shipping, and/or storage. In this work we have investigated the interactions between two non-ionic surfactants (Tween 20 and Tween 80) and bovine serum albumin (BSA), as model protein, using surface tension, fluorescence measurements and computational analysis. The results showed that, in both cases, the surface tension profile of the surfactants curve is modified upon addition of the protein, and the CMC values of Tween 20 and Tween 80 in the presence of protein are higher than the CMC values of the pure surfactants. The results indicate that although Tween 20 and Tween 80 do not greatly differ in their chemical structures, their interactions with BSA are of different nature, with distinct binding sites. Measurements at different protein concentrations showed that the interactions are also dependent on the protein aggregation state in solution. It was found from fluorescence studies that changes observed in both the intensity and wavelength of the tryptophan emission are probably caused by modifications of tryptophan environment due to surfactant binding, rather than by direct interaction. Based on a computational analysis of a BSA three-dimensional model, we hypothesize about the binding mechanism of non-ionic surfactant to globular protein, which allowed us to explain surface tension profiles and fluorescence results.