This study reports the production of multiple Acyl-Homoserine Lactones (AHLs), quorum sensing molecules thought to be exclusive to Gram-negative, in six Streptomyces species, prominent Gram-positive bacteria, challenging the previously established paradigm. These findings suggest the potential existence of a novel regulatory circuit in Streptomyces, organisms possessing significant secondary metabolism and complex life cycle. These findings hold substantial implications for the industrial, clinical, and evolutionary aspects of Streptomyces. The identification of a new potential regulatory circuit opens avenues for future investigations into the physiological roles of AHLs, the characterization of synthesizing enzymes, and the elucidation of molecular structures in Streptomyces and other Gram-positive bacteria.