AdpA, an AraC/XylS family protein, had been proved as a key regulator for secondary metabolism and morphological differentiation inStreptomyces griseus. Here, we identify AdpAch, an ortholog of AdpA, as a "higher level" pleiotropic regulator of natamycin biosynthesis with bidirectional regulatory ability inStreptomyces chattanoogensisL10. DNase I footprinting revealed six AdpAch-binding sites in thescnRI-scnRIIintergenic region. Further analysis using thexylEreporter gene fused to thescnRI-scnRIIintergenic region of mutated binding sites demonstrated that the expression ofscnRIandscnRIIwas under the control of AdpAch. AdpAchshowed a bi-stable regulatory ability where it firstly binds to the Site C and Site D to activate the transcription of the two pathway-specific genes,scnRIandscnRII, and then binds to other sites where it acts as an inhibitor. When Site A and Site F were mutatedin vivo, the production of natamycin was increased by 21% and 25%, respectively. These findings indicated an autoregulatory mechanism where AdpAchserves as a master switch with bidirectional regulation for natamycin biosynthesis.