This study aimed to investigate kojic acid production from the fermentation of rice by using Amylomyces rouxii TISTR 3182 and Aspergillus oryzae TISTR 3259. Steamed rice was fermented by A. rouxii followed by A. oryzae for 15 days, and mixed and separated cultures of both strains were compared. Kojic acid was analysed using Bentley's colorimetric method while glucose was analysed by the dinitrosalicylic acid (DNS) method. The results showed that separation of A. rouxii from rice medium before the addition of A. oryzae produced more kojic acid than the mixed culture, reaching a maximum yield of approximately 1.54 ± 0.10 g/l at day 10, pH 4 and 27 ± 2°C. The fermented broth was evaluated for antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, producing a maximum inhibition of 84 ± 11% and antityrosinase activity by dopachrome method with a maximum inhibition of 89 ± 11%. It may be concluded that microbial fermentation of rice could provide a good source of kojic acid. Separation of A. rouxii TISTR 3182 before the addition of A. oryzae TISTR 3259 in rice medium resulted in more effective kojic acid production. The fermented mixtures may have the potential to be further utilized as functional food and cosmetic ingredients, providing antioxidant and whitening activity.