LAUSR

Xanthomonadins are yellow pigments that are produced by the phytopathogen Xanthomonas campestris pv. campestris (Xcc). A pig cluster is responsible for xanthomonadin biosynthesis. Previously, Xcc4014 of the cluster was characterized as a bifunctional chorismatase that produces 3‐hydroxybenzoic acid (3‐HBA) and 4‐HBA. In this study, genetic analysis identified 11 genes within the pig cluster to be essential for xanthomonadin biosynthesis. Biochemical and bioinformatics analysis suggest that xanthomonadins are synthesized via an unusual type II polyketide synthase pathway. Heterologous expression of the pig cluster in non‐xanthomonadin‐producing Pseudomonas aeruginosa strain resulted in the synthesis of chlorinated xanthomonadin‐like pigments. Further analysis showed that xanC encodes an acyl carrier protein (ACP) while xanA2 encodes a ATP‐dependent 3‐HBA:ACP ligase. Both of them act together to catalyse the formation of 3‐HBA‐S‐ACP from 3‐HBA to initiate xanthomonadin biosynthesis. Finally, we showed that xanH encodes a FabG‐like enzyme and xanK encodes a novel glycosyltransferase. Both xanH and xanK are not only required for xanthomonadin biosynthesis, but also required for the balanced biosynthesis of extracellular polysaccharides and DSF‐family quorum sensing signals. These findings provide us with a better understanding of xanthomonadin biosynthetic mechanisms and directly demonstrate the presence of extensive cross‐talk among xanthomonadin biosynthetic pathways and other metabolic pathways.