A novel class I lanthipeptide produced by the marine bacterium Thalassomonas viridans XOM25T was identified using genome mining. The putative lanthipeptides were heterologously coexpressed in Escherichia coli as GFP-prepeptide fusions along with the operon-encoded class I lanthipeptide modification machinery VdsCB. The core peptides, VdsA1 and VdsA2, were liberated from GFP using the NisP protease, purified, and analyzed by collision-induced tandem mass spectrometry. The operon-encoded cyclase and dehydratase, VdsCB, exhibited lanthipeptide synthetase activity via post-translational modification of the VdsA1 and VdsA2 core peptides. Modifications were directed by the conserved double glycine leader containing prepeptides of VdsA1 and VdsA2.
Keywords: Escherichia coli heterologous expression; GFP fusion; Gram-negative; class I lanthipeptides; natural product discovery; viridisin.