Objective To investigate the effect of different doses of microbe-derived antioxidant on sleep and antioxidant ability in mice. Methods Sixty male Kunming mice with similar body weight were randomly divided into 4 groups.The control group received normal saline, and the experimental groups received microbe-derived antioxidant in a dose of 0.5 g/kg bw, 1.0 g/kg bw or 1.5 g/kg bw once per day, respectively. The experiment period was 30 days. At the end of experiment, the mice of each group were intraperitoneally injected sodium pentobarbital to induce sleep. The mice fall sleep was judged by righting reflex. After the test of sleep, blood was taken for detection of serum antioxidant ability. Results Compared with the low dose and high dose groups, the middle dose group showed a significantly prolonged sodium pentobarbital-induced sleeping time (P<0.05). Compared with the control group, low and high dose groups, the middle dose group had highly significantly increased GSH-Px activity (P<0.01) and significantly increased content of SOD. Under these conditions, the middle dose group reduced both the contents of MDA and 8-ISO-PGF2α (P<0.05) compared with the control group. Conclusions Our results suggest that microbe-derived antioxidant exerts effect on sleep and antioxidant ability in rats. Supplement of 1.0 g/kg bw/d shows the most significant effects.