ThnL, a B12-dependent radical S -adenosylmethionine enzyme, catalyzes thioether bond formation in carbapenem biosynthesis

Abstract

Complex carbapenems are important clinical antibiotics used to treat recalcitrant infections. Their biosynthetic gene clusters contain three essential B 12 -dependent radical S -adenosylmethionine (rSAM) enzymes. The majority of characterized enzymes in this subfamily catalyze methyl transfer, but only one is required to sequentially install all methionine-derived carbons in complex carbapenems. Therefore, it is probable that the other two rSAM enzymes have noncanonical functions. Through a series of fermentation and in vitro experiments, we show that ThnL uses radical SAM chemistry to catalyze thioether bond formation between C2 of a carbapenam precursor and pantetheine, uniting initial bicycle assembly common to all carbapenems with later tailoring events unique to complex carbapenems. ThnL also catalyzes reversible thiol/disulfide redox on pantetheine. Neither of these functions has been observed previously in a B 12 -dependent radical SAM enzyme. ThnL expands the known activity of this subclass of enzymes beyond carbon–carbon bond formation or rearrangement. It is also the only radical SAM enzyme currently known to catalyze carbon–sulfur bond formation with only an rSAM Fe–S cluster and no additional auxiliary clusters.