Streptomyces SBI034 produces several bafilomycin derivatives. Its afsA homologue (stcA) and putative gamma-butyrolactone receptor gene (stcB) were cloned. Construction of a stcA disruptant (stcA gene knockout) resulted in complete abolishment of all bafilomycin production. Electron microscopic analysis showed a defect of aerial mycelium formation and sporulation in the stcA disruptant. Restoration of all phenotypic defects and bafilomycin production was observed in a stcA complemented strain. Addition of exogenous gamma-butyrolactone (GBL) extracted from the culture broth of the wild-type strain could stimulate the aerial mycelium and spore formation of the stcA disruptant. These results suggest that stcA plays a role in GBL-mediated regulation of bafilomycin biosynthesis and morphological development in Streptomyces strain SBI034.