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Keywords:

  • natural products;
  • Photorhabdus;
  • protein expression;
  • secondary metabolites;
  • Xenorhabdus;
  • yeast homologous recombination cloning

Abstract

Heterologous production of GameXPeptide A (1), as well as of the novel peptide natural products ambactin (2) and xenolindicins A–C (3 ac), 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.