Optimization of purification process and thermal stability of lactoferrin from camel milk

To expand the application of camel lactoferrin (CLF) in thermally processed foods, this study optimized cation-exchange chromatography to investigate its thermally induced conformational evolution. Under optimal conditions (flow rate 1 mL/min, pH 7.0, impurity elution salt concentration containing 0.3 M NaCl), CLF with purity of 90.13% ± 1.06% was obtained with good repeatability (RSD < 5%) for preparation and quantitative analysis. Compared with bovine lactoferrin (BLF), CLF exhibits divergent lobe thermostability. Fluorescence and UV-vis spectra confirm severer unfolding and hydrophobic exposure in CLF at 95 °C, whereas DLS reveals weaker aggregation, showing inconsistent trends between unfolding and aggregation. Molecular dynamics simulations demonstrate CLF adopts a compact internally disordered conformation. Strengthened intramolecular salt bridges and hydrophobic interactions shield hydrophobic sites and inhibit aggregation. This study provides process-optimization ideas and structural insights for the application of CLF in thermally processed foods and the high-value utilization of camel milk resources.