Distiller's grain (DG), a byproduct of the Chinese alcohol industry, can be used as a renewable organic waste resource for biofuel through pyrolysis. However, amino acids and protein are the main component of DG and have obvious effects on its pyrolysis. In this study, the properties of DG and extracted DG (EDG) and their pyrolysis behaviors were compared in the temperature range 50−900 °C. 3D-FTIR revealed that the main gaseous products of DG were CO 2 , CH 4 , ketones, aldehydes, acids, and amines, and that the deamination of amino acids and protein was inhibited by lignocellulosic components during DG pyrolysis. The TG/DTG results showed that DG could be pyrolyzed at a lower temperature, and the absolute value of maximum DTG loss rate was 6% min −1 , higher than that of EDG. The distributed activation energy model (DAEM) was used to analyze the kinetic behaviors of DG and EDG. Meanwhile, the Flynn−Wall−Ozawa (FWO) method was also used to confirm the results of DAEM, and the results proved that the existence of soluble components can reduce the reaction barrier and enhance the reaction rate of DG. This study provides a basic reference for the full utilization of DG through the pyrolysis process.