The interaction between L-Tryptophan (L-TRP) and chlorogenic acid (CGA) was studied using UV/Vis absorption spectra, steady-state fluorescence, synchronous fluorescence spectra, Time-resolved fluorescence spectra, Fourier transform Infrared (FTIR), antibacterial activity, and molecular docking. The fluorescence of L-Tryptophan is exquisitely sensitive to the environment. It’s sensitivity is related to luminescence properties originating from the hetero aromatic indole - ring of the residue with an uneven charge distribution around the macrocycle. Chlorogenic acid is the most abundant hydroxycinnamic acid naturally available in beverages of plant origin, such as coffee, and other agricultural product such as fruits, vegetables, cereals, and legumes. The conformation changes have been noted and the quenching mechanism between (L-TRP) and CGA proved to be static quenching by fluorescence. The binding constant and the number of the binding sites (L-TRP) on CGA were also calculated in this study.