Yeast Homologous Recombination Cloning Leading to the Novel Peptides Ambactin and Xenolindicin

Abstract Heterologous production of GameXPeptide A ( 1 ), as well as of the novel peptide natural products ambactin ( 2 ) and xenolindicins A–C ( 3 a – c ), was achieved by using the “overlap extension PCR‐yeast homologous recombination” (ExRec) method. ExRec cloning is based on the ability of yeast to assemble overlapping DNA fragments into functional plasmids. Here we used this technique to clone a total of 15 biosynthesis gene clusters from Photorhabdus and Xenorhabdus with sizes of up to 45 kb. The structures of the novel compounds 2 and 3 a , which were produced in Escherichia coli , were elucidated by detailed MS and bioinformatics analysis, and additionally confirmed by their chemical synthesis.