Probing the binding properties of dicyandiamide with pepsin by spectroscopy and docking methods.

Dicyandiamide (DCD), considered to be a nitrification inhibitor, poses threat to human's health with exposure from milk, infant formula and other food products. In this work, DCD was investigated for its binding reaction with pepsin using spectroscopy and docking methods. Fluorescence experiments indicated DCD quenched the fluorescence of pepsin through a static process. Thermodynamic analysis of the binding data (ΔH0 = -21.72 kJ mol-1 and ΔS0 = 17.61 J mol-1 K-1) suggested the involvement of hydrophobic and hydrogen bonding in the complex formation. The pepsin interacted with DCD at a hydrophobic cavity, leading to a conformational changes in the pepsin, as revealed from UV-vis absorption, Fourier transform infrared, the time-resolved fluorescence, three-dimensional fluorescence and circular dichroism spectral results.