The effect of active oxygen species on the productivity of torularhodin, an effective antioxidant, by soil yeast, Rhodotorula glutinis no. 21, was examined. Methylene blue, methyviologen and AAPH [2,2'-azobis(2-amidinopropane) dihydrochloride] were used as generators of singlet oxygen, superoxide anion radicals and peroxy radicals, respectively. All of them indicated effectiveness at a dose of 1.0 x 10(-10) to 3.0 x 10(-6) M. Addition of these generators to the culture medium had almost no influence on the biosynthesis of beta-carotene, whereas it had marked enhancement on those of torulene and torularhodin. Production of uric acid by xanthineoxidase remained unchanged at a torularhodin concentration of up to 100 microM. This result suggests that torularhodin does not directly affect the productivity of superoxide anions. It has been proved that torularhodin has a more potent effect on the scavenging of peroxyl radicals and inhibits substrate degradation by singlet oxygen more effectively than beta-carotene does. Continuous addition of methylene blue enhanced the torularhodin accumulation more markedly than single addition.