Pyridoxal-5′-phosphate-dependent enzyme GenB3 Catalyzes C-3′,4′-dideoxygenation in gentamicin biosynthesis

Abstract Background The C-3′,4′-dideoxygenation structure in gentamicin can prevent deactivation by aminoglycoside 3′-phosphotransferase (APH(3′)) drug-resistant pathogens. However, the enzyme catalyzing dideoxygenation step biosynthesis pathway remains unknown. Results Here, we report that GenP catalyzes 3′ phosphorylation of intermediates JI-20A, JI-20Ba, and JI-20B. We further demonstrate pyridoxal-5′-phosphate (PLP)-dependent GenB3 uses these phosphorylated substrates to form 3′,4′-dideoxy-4′,5′-ene-6′-oxo products. following C-6′-transamination GenB4-catalyzed reduction 4′,5′-olefin lead formation C. To best our knowledge, is first PLP-dependent biosynthesis. Conclusions This discovery solves a long-standing puzzle enriches knowledge chemistry enzymes. Interestingly, results evade APH(3′) pathogens, producers evolved smart strategy, which utilized their own activate hydroxyls as leaving groups for 3′,4′-dideoxygenation