title: Heterologous expression of a myxobacterial natural products assembly line in pseudomonads via red/ET recombineering.
abstract: Natural products of microbial origin are widely used as pharmaceuticals and in agrochemistry. These compounds are often biosynthesized by multifunctional megasynthetases whose genetic engineering and heterologous expression offer considerable promise, especially if the natural hosts are genetically difficult to handle, slow growing, unculturable, or even unknown. We describe a straightforward strategy that combines the power of advanced DNA engineering (recombiogenic cloning) in Escherichia coli with the utility of pseudomonads as the heterologous host for the analysis and mutagenesis of known and unknown secondary metabolite pathways. The myxochromide S biosynthetic gene cluster from Stigmatella aurantiaca was rebuilt and engineered in E. coli to contain the elements required for expression in pseudomonads. The successful production in Pseudomonas putida, at unprecedented levels, demonstrates the feasibility of the new approach to the analysis and mutagenesis of these important pathways.
authors.affiliation: Pharmaceutical Biotechnology, Saarland University, P.O. Box 151150, 66041 Saarbrücken, Germany.
authors.forename: Silke C
authors.initials: SC
authors.lastname: Wenzel
authors.forename: Frank
authors.initials: F
authors.lastname: Gross
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.forename: Jun
authors.initials: J
authors.lastname: Fu
authors.forename: A Francis
authors.initials: AF
authors.lastname: Stewart
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
mesh_terms: D004274:DNA, Recombinant / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D018014:Gene Transfer Techniques; D005798:Genes, Bacterial / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics*; D005810:Multigene Family / Q000502:physiology; D009231:Myxococcales / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000235:genetics*; D011549:Pseudomonas / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D000097573:Red Fluorescent Protein
chemical_list: D004274:DNA, Recombinant; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein