title: 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.
authors.affiliation: Merck Stiftungsprofessur für Molekulare Biotechnologie, Fachbereich Biowissenschaften, Goethe Universität Frankfurt, Max-von-Laue Strasse 9, 60438 Frankfurt am Main (Germany).
authors.forename: Olivia
authors.initials: O
authors.lastname: Schimming
authors.forename: Florian
authors.initials: F
authors.lastname: Fleischhacker
authors.forename: Friederike I
authors.initials: FI
authors.lastname: Nollmann
authors.forename: Helge B
authors.initials: HB
authors.lastname: Bode
mesh_terms: D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D059765:Homologous Recombination; D008968:Molecular Conformation; D009842:Oligopeptides / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D010456:Peptides, Cyclic / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D020637:Photorhabdus / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D020593:Xenorhabdus / Q000378:metabolism
chemical_list: D009842:Oligopeptides; D010456:Peptides, Cyclic
keywords: Photorhabdus; Xenorhabdus; natural products; protein expression; secondary metabolites; yeast homologous recombination cloning