title: Functional characterization of different ORFs including luciferase-like monooxygenase genes from the mensacarcin gene cluster.
abstract: The biologically active compound mensacarcin is produced by Streptomyces bottropensis. The cosmid cos2 contains a large part of the mensacarcin biosynthesis gene cluster. Heterologous expression of this cosmid in Streptomyces albus J1074 led to the production of the intermediate didesmethylmensacarcin (DDMM). In order to gain more insights into the biosynthesis, gene inactivation experiments were carried out by λ-Red/ET-mediated recombination, and the deletion mutants were introduced into the host S. albus. In total, 23 genes were inactivated. Analysis of the metabolic profiles of the mutant strains showed the complete collapse of DDMM biosynthesis, but upon overexpression of the SARP regulatory gene msnR1 in each mutant new intermediates were detected. The compounds were isolated, and their structures were elucidated. Based on the results the specific functions of several enzymes were determined, and a pathway for mensacarcin biosynthesis is proposed.
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany).
authors.forename: Sarah
authors.initials: S
authors.lastname: Maier
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany).
authors.forename: Tanja
authors.initials: T
authors.lastname: Heitzler
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany).
authors.forename: Katharina
authors.initials: K
authors.lastname: Asmus
authors.affiliation: Organic Chemsitry II, Universität Siegen, Adolf-Reichwein-Strasse 2, 57068 Siegen (Germany).|Present address: Helmholtz Institut für Pharmazeutische Forschung Saarland, Postfach 151150, 66041 Saarbrücken (Germany).
authors.forename: Elke
authors.initials: E
authors.lastname: Brötz
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany).
authors.forename: Uwe
authors.initials: U
authors.lastname: Hardter
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany).
authors.forename: Katharina
authors.initials: K
authors.lastname: Hesselbach
authors.affiliation: Organic Chemsitry II, Universität Siegen, Adolf-Reichwein-Strasse 2, 57068 Siegen (Germany).
authors.forename: Thomas
authors.initials: T
authors.lastname: Paululat
authors.affiliation: Institut für Pharmazeutische Biologie und Biotechnologie, Albert-Ludwigs Universität, Stefan-Meier-Strasse 19, 79104 Freiburg (Germany). andreas.bechthold@pharamzie.uni-freiburg.de.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bechthold
mesh_terms: D000873:Anthracenes / Q000378:metabolism*; D055162:Biocatalysis; D017353:Gene Deletion; D008156:Luciferases / Q000235:genetics*; D006899:Mixed Function Oxygenases / Q000172:deficiency / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D005810:Multigene Family / Q000235:genetics*; D016366:Open Reading Frames / Q000235:genetics*; D013302:Streptomyces / Q000201:enzymology / Q000235:genetics / Q000378:metabolism
chemical_list: D000873:Anthracenes; C510239:mensacarcin; D006899:Mixed Function Oxygenases; D008156:Luciferases
keywords: biosynthesis; didesoxymensacarcin; mensacarcin; polyketides; structure elucidation