Discovery and heterologous expression of microginins from Microcystis aeruginosa LEGE 91341

Microginins are a large family of cyanobacterial lipopeptide protease inhibitors. A hybrid polyketide synthase (PKS)/non-ribosomoal peptide synthetase (NRPS) biosynthetic gene cluster (BGC) found in several microginin-producing strains – mic – was proposed to encode the production of microginins, based on bioinformatic analysis. Here, we explored a cyanobacte-rium, Microcystis aeruginosa LEGE 91341, which contains a mic BGC, to discover twelve new microginin variants. The new compounds contain uncommon amino acids, namely homophenylalanine (Hphe), homotyrosine (Htyr) or methylproline, as well as a 3-aminodecanoic acid (Ada) residue, which in some variants was chlorinated at its terminal methyl group. We have used Direct Pathway Cloning (DiPaC) to heterologously express the mic BGC from M. aeruginosa LEGE 91341 in E. coli, which led to the production of several microginins. This proved that the mic BGC is, in fact, responsible for the biosynthesis of microginins and paves the way to accessing new variants from (meta)genome data or through pathway engineering.