<p><i>Catharanthus roseus</i> is an important medicinal plant and the only source of the important anticancer compounds vincristine and vinblastine. Elicitors are compounds that stimulate plant defense systems by enhancing the immune response through increasing the activity of defense enzymes and the production of defense compounds, as well as simultaneously increasing the production of secondary metabolites through the elicitation of their metabolic pathways. In present study, mycelium extract (CE) and culture filtrate (CF) of endophytic fungus <i>Schizophyllum commune</i> were used as elicitors at concentrations (0, 2, 4, and 6% v/v) at two 48 and 72 hours’ exposure times to induce antioxidant enzyme activity and phenolic compounds production in the <i>C. roseus</i> cell suspension. Results showed that 4% CF at 48&#xa0;h yielded the highest total phenol content (19.28 ± 0.385&#xa0;mg GAE g^− 1 FW), while 4% CE at 48 produced the highest total flavonoid content (6.93 ± 0.265&#xa0;mg QUE g^− 1 FW) and antioxidant activity (25.83 ± 0.925%DPPH inhibition). Quercetin decreased in CF and CE treated, and control cells had the highest quercetin. Cinnamic acid was detected only in CF treated, and apigenin was identified only with CE at 48&#xa0;h. 6% CF at 48&#xa0;h and 2% CE at 72 hours’ exposure time produced the highest gallic acid (2.52 ± 0.002&#xa0;µg g^− 1 FW) and rosmarinic acid (0.49 ± 0.003&#xa0;µg g^− 1 FW) content, respectively. The highest catalase (1.79 ± 0.100 U g^− 1 FW) and guaiacol peroxidase (0.084 ± 0.002 U g^− 1 FW) activity was with 2% and 6% CF for 72&#xa0;h, respectively. Ascorbate peroxidase activity generally decreased below control levels, except in 4% CE for 78&#xa0;h its activity increased slightly and reached the control levels. In conclusion, our results show that CF demonstrates a stronger elicitor than CE, particularly at higher concentrations and longer exposure, for inducing defense responses in <i>C. roseus</i> cell suspension culture.</p>