Biological buffers are always considered as non-toxic, biocompatible and green compounds. Therefore, we analyzed the catalytic activity of a commercial enzyme α-chymotrypsin (α-CT) in aqueous solutions of some common biological buffers (TRIS, TES, TAPS, and TAPSO) at pH 8 and T = 25 °C. It is found that the increase of the buffer concentration enhanced the catalytic activity of enzyme α-CT, and the tendency follows the order of TRIS > TES > TAPS > TAPSO. Especially in the presence of TRIS, the catalytic activity enhanced 5.5 fold.