The present study aims to evaluate the levels of acetyl cholinesterase activity, specific activity, and total protein in blood samples obtained from persons who have received a diagnosis of autism spectrum disorder. Furthermore, the objective of this study is to determine the most favourable parameters for acetyl cholinesterase concentration, substrate concentration, pH level, temperature, and duration. In addition, the present study aims to ascertain the Km and Vmax values of acetyl cholinesterase in blood samples obtained from patients diagnosed with autism spectrum disorder. A complete collection of 100 samples was procured from individuals who have been diagnosed with autism spectrum disorder (ASD). The study's results suggest that persons diagnosed with ASD demonstrate reduced levels of enzyme activity in comparison to the control group. Moreover, the enzymatic activity in individuals with ASD is shown to be significantly elevated compared to the control group. In contrast to the control group, individuals diagnosed with ASD exhibit decreased amounts of total protein. persons with ASD exhibit slight deviations in the thermodynamic characteristics of enzymes, including the activation energy, Delta G, Delta S, Pz, and Delta H*, when compared to persons in the control group.