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Interactions of hyaluronan with micelles formed by cationic surfactants were studied by the time-resolved measurement of fluorescence resonance energy transfer (FRET) using perylene and fluorescein as probes. Two surfactants were studied – Cetyltrimethylammonium bromide (CTAB) and Septonex. In pure micellar solutions, the same values of FRET efficiency were found for both surfactants, but values for the binding of the probe pair were lower for Septonex micelles than in the case of CTAB. This was attributed to steric effects of the carbethoxy group in the Septonex polar head. Upon the addition of hyaluronan, decreased FRET efficiency and increased binding were detected in comparison with pure surfactants. To resolve the structure of the formed aggregates, steady state and time-resolved fluorescence anisotropy was employed as an additional technique. All results indicated that cationic micelles bind to hyaluronan forming a pearl necklace structure with micelles of smaller size compared to pure surfactant. Besides theoretical interest, the studied polyelectrolyte-surfactant system may be interesting for the formulation of drug delivery systems. [Display omitted] • Cetyltrimethylammonium bromide and Septonex show comparable behavior when interacting with the biopolymer hyaluronan. • The differences in the resulting aggregates are due to the properties of the starting surfactants. • The proposed aggregation model for this interaction is the pearl necklace structure. [ABSTRACT FROM AUTHOR]