Mechanistic insights into ammonium-driven metabolic regulation for enhanced nemadectin biosynthesis in Streptomyces cyaneogriseus

Abstract Nemadectin, a milbemycin-class macrocyclic lactone antibiotic produced by Streptomyces cyaneogriseus, is a potent broad-spectrum insecticide with excellent environmental compatibility. Its derivative moxidectin, featuring a C-23 methoxime modification, demonstrates enhanced insecticidal activity and has become a commercially successful agrochemical. This study reveals ammonium regulation effectively boosts nemadectin biosynthesis in S. cyaneogriseus, with mechanistic insights gained through integrated multi-omics analysis. Transcriptomic profiling showed ammonium sulfate supplementation significantly upregulates the nemadectin biosynthetic gene cluster, including polyketide synthase (PKS) genes, backbone modification genes, and pathway-specific transcription factors, while also enhancing the expression of Avenolide-like signaling molecules and global transcription factor Afskne. Metabolomic dynamics revealed reinforced precursor biosynthesis through coordinated metabolic reprogramming: enhanced acetyl-CoA production, reinforced Embden–Meyerhof–Parnas pathway and amino acid/acyl-CoA metabolism, coupled with reduced tricarboxylic acid cycle activity. Systematic integration of physiological phenotyping, metabolite profiling, and transcriptional regulation data comprehensively elucidated the ammonium-driven overproduction mechanism, providing critical insights for developing advanced fermentation strategies and genetic engineering approaches in industrial antibiotic production. Graphical abstract