Neurospora intermedia N-1 is a fungus isolated from “oncom merah”, an Indonesian red peanut cake. It has been studied for carotenoids production for food and drug colorant. Commercial production of natural carotenoids from microorganisms recently competes mainly with synthetic carotenoids by chemical procedures. Improving the efficiency of carotenoid biosynthesis can increase carotenoids production by microorganisms. Some of the cultural and environmental factors are positively affect carotenoids content of the fungal strains. The objective of this study was to evaluate the carotenogenesis of Neurospora intermedia N-1 in a liquid fermentation. The fermentation was started by optimizing the condition with light responses, then addition of variated carbon sources at 2 % w/v, cofactor Mg2+, antibiotics penicillin and chloramphenicol. Carotenoid analysis was carried out by spectrophotometer UV/Vis at 480 nm and HPLC using a C18 column with acetonitrile: methanol: 2-propanol (85:10:5) as the mobile phase for isocratic elution. The research result showed that fermentation within 3 days incubation in dark condition and 4 days under blue light was better than 7 days in dark condition, and produced carotenoid 8.13 μg/g spores. Substrate with addition of maltose 2 % w/v and incubation at aerobic condition produced carotenoid 19.913 μg/g spores. Stimulation by co-factor Mg2+ in the ranges concentration below 10 mM increased the total carotenoids up to 24.31 μg/g spores, but decreased when was stimulated higher than 12 mM. Stimulation by antibiotics penicillin and chloramphenicol at concentration of 1 mg/mL increased the β-carotene content. Penicillin influenced more significant to the carotenogenesis of Neurospora intermedia N-1, particularly for β-carotene production up to 17.193 μg/g spores.