Puerarin, which was used as a natural isoflavone in this study, was obtained from the Kudzu plant. The inhibitory effect of puerarin on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) enzymes was investigated. The IC50 values of AChE (177.73 nM) and BChE (315.07 nM) were determined. The binding mechanism of puerarin, which has a high inhibitory effect on the active site of the enzyme, was demonstrated by the molecular docking method. The antioxidant activity was evaluated for the same molecule. For this purpose, Fe3+‐Fe2+ reducing capacity, ABTS radical scavenging activity, CUPRAC method, DPPH⋅ scavenging activity and DMPD radical scavenging method were used. The percentage (%) inhibition amount of puerarin molecule was calculated as 99.33 % for ABTS radical scavenging activity, which is the more preferred method for natural products. The absorbtion value of iron reduction of Puerarin was 0.122±0.010, and the value of copper reduction was 0.633±0.026. Our research efforts include uncovering the potent antioxidant potential of puerarin and, simultaneously, a comparative study of tacrine, primarily emphasizing its inhibitory effects against AChE and BChE enzymes. The puerarin is expected to be a guide for drug discovery against many problems, especially oxidative cell damage in Alzheimer's disease, mitochondrial dysfunction (MD), cancer, and cardiovascular diseases.