Silver nanoparticles (AgNPs) functionalized with glutamic acid (GA) was used as a biochemical sensing probe in colorimetry for detection of lysozyme protein in milk samples. The method is based on the color change of AgNPs/GA from yellow to reddish-yellow differentiated with naked eyes for qualitative determination and red shift of localized surface plasmon resonance (LSPR) absorption signal intensity of AgNPs/GA in visible region used for quantitative determination of lysozyme. The control experiments were performed to demonstrate the electrostatic force of interactions between AgNPs/GA and protein molecule. A wide linear range of 3-150 nM with limit of detection of 1.5 nM was acquired for quantitative determination of lysozyme using AgNPs/GA as a biochemical sensing probe. The advantages of using AgNPs/GA as a biochemical sensing probe are simple, label-free and economic for determination of lysozyme from milk samples.