title: Reconstitution of the myxothiazol biosynthetic gene cluster by Red/ET recombination and heterologous expression in Myxococcus xanthus.
abstract: Although many secondary metabolites exhibiting important pharmaceutical and agrochemical activities have been isolated from myxobacteria, most of these microorganisms remain difficult to handle genetically. To utilize their metabolic potential, heterologous expression methodologies are currently being developed. Here, the Red/ET recombination technology was used to perform all required gene cluster engineering steps in Escherichia coli prior to the transfer into the chromosome of the heterologous host. We describe the integration of the complete 57-kbp myxothiazol biosynthetic gene cluster reconstituted from two cosmids from a cosmid library of the myxobacterium Stigmatella aurantiaca DW4-3/1 into the chromosome of the thus far best-characterized myxobacterium, Myxococcus xanthus, in one step. The successful integration and expression of the myxothiazol biosynthetic genes in M. xanthus results in the production of myxothiazol in yields comparable to the natural producer strain.
authors.affiliation: Institut für Pharmazeutische Biotechnologie, Universität des Saarlandes, Postfach 15 11 50, D-66041 Saarbrücken, Germany.
authors.forename: Olena
authors.initials: O
authors.lastname: Perlova
authors.forename: Jun
authors.initials: J
authors.lastname: Fu
authors.forename: Silvia
authors.initials: S
authors.lastname: Kuhlmann
authors.forename: Daniel
authors.initials: D
authors.lastname: Krug
authors.forename: A Francis
authors.initials: AF
authors.lastname: Stewart
authors.forename: Youming
authors.initials: Y
authors.lastname: Zhang
authors.forename: Rolf
authors.initials: R
authors.lastname: Müller
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001709:Biotechnology / Q000379:methods; D016887:Cardiopulmonary Resuscitation; D003001:Cloning, Molecular; D003360:Cosmids; D018274:Electroporation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D008689:Methacrylates / Q000378:metabolism; D005810:Multigene Family; D016941:Myxococcus xanthus / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D011995:Recombination, Genetic; D020597:Stigmatella / Q000201:enzymology* / Q000235:genetics; D013844:Thiazoles / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D008689:Methacrylates; D013844:Thiazoles; C030517:myxothiazol