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AdpA, an AraC/XylS family protein, had been proved as a key regulator for secondary metabolism and morphological differentiation in  Streptomyces griseus . Here, we identify AdpA ch , an ortholog of AdpA, as a “higher level” pleiotropic regulator of natamycin biosynthesis with bidirectional regulatory ability in  Streptomyces chattanoogensis  L10. DNase I footprinting revealed six AdpA ch -binding sites in the  scnRI – scnRII  intergenic region. Further analysis using the  xylE  reporter gene fused to the  scnRI – scnRII  intergenic region of mutated binding sites demonstrated that the expression of  scnRI  and  scnRII  was under the control of AdpA ch . AdpA ch  showed 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,  scnRI  and  scnRII , and then binds to other sites where it acts as an inhibitor. When Site A and Site F were mutated  in vivo , the production of natamycin was increased by 21% and 25%, respectively. These findings indicated an autoregulatory mechanism where AdpA ch  serves as a master switch with bidirectional regulation for natamycin biosynthesis.

Bidirectional Regulation of AdpAch in Controlling the Expression of scnRI and scnRII in the Natamycin Biosynthesis of Streptomyces chattanoogensis L10