Gene Cluster Directs Diversified Carbazole Biosynthesis via Cooperative Halogenation, Hydroxylation, and Methylation.

Chlocarbazomycins are bioactive carbazoles with therapeutic potential. Here, we report a conserved gene cluster (ccz) that directs its biosynthesis via a novel pathway. This cluster encodes a versatile tailoring system, including a regioselective halogenase (CczH), dedicated hydroxylases (CczA/J/D), and a bifunctional methyltransferase (CczI) that iteratively methylates and accelerates N-OH dehydroxylation. Our findings provide new enzymes for carbazole pathway engineering and structural diversification.