A Defined and Flexible Pocket Explains Aryl Substrate Promiscuity of the Cahuitamycin Starter Unit–Activating Enzyme CahJ

Abstract

Cahuitamycins are biofilm inhibitors assembled by a convergent nonribosomal peptide synthetase pathway. Previous genetic analysis indicated that a discrete enzyme, CahJ, serves as a gatekeeper for cahuitamycin structural diversification. Here, the CahJ protein was probed structurally and functionally to guide the formation of new analogues by mutasynthetic studies. This analysis enabled the in vivo production of a new cahuitamycin congener through targeted precursor incorporation.Breaking the barrier: Biofilm formation is employed by pathogenic microbes to defend against antibiotic action. This study probes both structurally and functionally CahJ, a key biosynthetic adenylation enzyme involved in generation of the cahuitamycin biofilm inhibitors, and lays a foundation for the development of effective new analogues.