Supplementation with rice oil and its major components (oleic acid and linoleic acid) was found to have a significant influence on cephalosporin C (CPC) production and cell growth by A. chrysogenum M35 in shake flask cultures. Five percent (v/v) rice oil had the most robust effect and 5% (v/v) oleic acid was the second most efficient on cell growth, whereas 3% (v/v) linoleic acid was found to be optimal for CPC production. Rice oil, oleic acid, and linoleic acid also significantly improved the rates of glucose consumption. When glucose was almost consumed, CPC production was initiated and, on the addition of rice oil, lipase activity increased steadily to 1.56 U/ml for 4 days. These results suggest that rice oil and fatty acids are used as carbon source to produce CPC by A. chrysogenum M35. Moreover, a mixture, composed of 40% (v/v) oleic acid and 60% (v/v) linoleic acid, had the strongest stimulatory effect on CPC production, due to a synergistic effect of the two fatty acids. Consequently, the maximum CPC titer (7.44 g/l) was improved about 4.5-fold.