Molecular insights on the binding of chlortetracycline to bovine casein and its effect on the thermostability of chlortetracycline.

[Display omitted] • The spontaneous binding behavior of CTC to casein was revealed for the first time. • Binding sites engaged in the formation of the casein-CTC complex were deciphered. • The complexation between casein and CTC inhibited the thermal degradation of CTC. Bovine casein was selected as a model protein to evaluate the impact of food matrix on the thermal degradation of antibiotics. Fluorescence quenching and isothermal titration calorimetry experiments revealed that chlortetracycline (CTC) could spontaneously bind to casein via hydrogen bonding and hydrophobic interactions. The amino acid residues forming the binding pocket were further identified using molecular docking, while saturation transfer difference NMR deciphered that the binding of CTC engages its -N(CH 3) 2 group. Moreover, the degradation behavior of free CTC versus that bound in casein-CTC complex was compared during thermal treatment. Compared with free CTC, a lower first-order rate constant was observed in the presence of casein. Removal of casein shortened the half-life of CTC by at least 48.1% at low concentrations. Elucidating that the formation of protein-antibiotic complexes alters the amenability of antibiotics to degradative reactions, which could help eliminate residual antibiotics and guarantee the safety of dairy products. [ABSTRACT FROM AUTHOR]