Identification of a novel butenolide signal system to regulate high production of tylosin in Streptomyces fradiae

Identifying hormone-like quorum sensing (QS) molecules in streptomycetes is challenging due to low production levels but is essential for understanding secondary metabolite biosynthesis and morphological differentiation. This work reports the discovery of a novel γ-butenolide-type signaling molecule (SFB1) via overexpressing its biosynthetic gene (orf18) in Streptomyces fradiae. SFB1 was found to be essential for production of tylosin through dissociating the binding of its receptor TylP (a transcriptional repressor) to target genes, thus activating the expression of tylosin biosynthetic gene cluster (tyl). Meanwhile, SFB1 biosynthesis is negatively regulated by TylQ (another transcriptional repressor); the disruption of its coding gene tylQ led to increased production of SFB1, which in turn increased the yield of tylosin. Using tylQ disrupted mutant as chassis cell, co-overexpressing transcriptional activators TylR and TylS further increased tylosin yield to 3926 ± 110 mg/L, representing a 2.93-fold improvement over the wild-type strain. Since the quorum sensing signaling system can affect the biosynthesis of many secondary metabolites, thereby this strategy may also be readily applied for improving the titers of other microbial metabolites. KEY POINTS: • SFB1 is a novel γ-butenolide-type quorum sensing signaling molecule of S. fradiae. • SFB1 regulates the production of tylosin. • Engineering SFB1 regulatory cascade improves tylosin production.