Streptomyces clavuligerus produces a clinically important beta-lactamase inhibitor, clavulanic acid. When several of the selected redox-cycling agents were treated, an increase in clavulanate production was observed. The stimulatory effect was seen when the reaction was fed with menadione, plumbagin and phenazine methosulfate (PMS), whereas feeding with methyl viologen had a negative effect. PMS exerted the strongest effect, enhancing the accumulation of clavulanic acid by 150%. Induction of superoxide dismutase upon the addition of PMS suggested an involvement of superoxide in the enhancing process. The stimulatory effect of PMS was offset by the addition of butylated hydroxyanisole, further supporting the involvement of the active oxygen. The enhanced production of clavulanic acid correlated well with the increased total activity of clavaminic acid synthase, a key enzyme in its biosynthesis, and the transcription of cas2, its coding gene. The results suggested that active oxygen species could enhance the biosynthesis of secondary metabolites through the transcriptional activation of the biosynthetic gene.