Lactoferrin/κ-Carrageenan Interactions: Thermodynamic and Kinetic Insights through Surface Plasmon Resonance and Steady-State Fluorescence.

Protein-polysaccharide complexes play important roles in a wide range of technological applications. To elucidate the molecular interactions governing the formation of bovine lactoferrin (BLF)-κ-carrageenan (κCG) bionanostructures, the thermodynamic and kinetic parameters of BLF-κCG binding were determined using Steady-state fluorescence (FS) and surface plasmon resonance (SPR) spectroscopies. Formation of a 1:1 BLF-κCG complex is thermodynamically favorable at equilibrium (-47.00 ≤ ΔG FS o ≤ -46.15 kJ mol-1) and is predominantly enthalpy-driven (ΔH FS o = -57.43 kJ mol-1). Kinetic analysis by SPR revealed that the interaction proceeds through a transition complex ([BLF - κCG]‡), with complex formation occurring more rapidly through association of the free molecules (32.53 ≤ ΔG a ‡ ≤ 38.52 kJ mol-1) than through dissociation of the thermodynamically stable complex (77.61 ≤ ΔG d ‡ ≤ 81.62 kJ mol-1). Consistent with these findings, the energetic barrier for dissociation (E d-298.2 K ‡ = 96.46 kJ mol-1) was higher than for association (E a-298.2 K ‡ = 42.88 kJ mol-1). Increasing the ionic strength altered the BLF-κCG interaction mechanism, converting the binding process from a multistep pathway to a single-step process.