A triclosan‐resistance protein from the soil metagenome is a novel enoyl‐acyl carrier protein reductase: Structure‐guided functional analysis

The synthetic biocide triclosan targets enoyl‐acyl carrier protein reductase(s) (ENR) in bacterial type II fatty acid biosynthesis. Screening and sequence analyses of the triclosan resistome from the soil metagenome identified a variety of triclosan‐resistance ENRs. Interestingly, the mode of triclosan resistance by one hypothetical protein was elusive, mainly due to a lack of sequence similarity with other proteins that mediate triclosan resistance. Here, we carried out a structure‐based function prediction of the hypothetical protein, herein referred to as FabMG, and in vivo and in vitro functional analyses. The crystal structure of FabMG showed limited structural homology with FabG and FabI, which are also involved in type II fatty acid synthesis. In vivo complementation and in vitro activity assays indicated that FabMG is functionally a FabI‐type ENR that employs NADH as a coenzyme. Variations in the sequence and structure of FabMG are likely responsible for inefficient binding of triclosan, resulting in triclosan resistance. These data unravel a previously uncharacterized FabMG, which is prevalent in various microbes in triclosan‐contaminated environments and provide mechanistic insight into triclosan resistance. The synthetic biocide triclosan targets and inhibits FabI, an enoyl‐acyl carrier protein reductase (ENR) involved in bacterial type II fatty acid synthesis. Here, we provide structural and functional evidences that a triclosan‐resistance enzyme FabMG from the soil metagenome is functionally a FabI‐type ENR, but variations in the sequence and structure of FabMG could be responsible for inefficient binding of triclosan, resulting in triclosan resistance.