Serratiopeptidase exhibits therapeutic efficacy in a variety of disease models; in addition to its ability to mediate anti-inflammatory action, it is also responsible for regulating several biological activities by targeting different signaling pathways. Our study aimed to evaluate the central nervous effect of serratiopeptidase on neurobehavioral activities and to eliminate anxiety manifestations in mice. Male mice were subjected to neurobehavioral tests, including open field, negative geotaxis, head poking, and swimming tests, in addition to methods of screening for antianxiety, such as elevated pulse maze and light-dark box tests, after one hour of serratiopeptidase at 5, 10, and 20 mg/kg orally. Serratiopeptidase at 20 mg/kg produced a significant increase in the number of squares cut and rearing compared to the control group. Serratiopeptidase at 5, 10, and 20 mg/kg resulted in a decrease in the time required to correct the position of mice in comparison with the control group in negative geotaxis, and there was an increase in the number of stockings compared with the control group. Serratiopeptidase (20 mg/kg) significantly increased the duration of time spent in the open arm and significantly decreased the amount of time spent in the closed arm compared with the control group and the light-dark box test. Serratiopeptidase at 20 mg/kg significantly increased the time spent on the light side of mice and significantly decreased the time spent on the dark side. Serratiopeptidase at 20 mg/kg demonstrated superior effects to the group treated with sertraline in reducing anxiety-like behavior in both tests. In conclusion, serratiopeptidase has a stimulatory effect on the central nervous system, and a high dose may produce anxiolytic-like effects.