title: An artificial pathway to 3,4-dihydroxybenzoic acid allows generation of new aminocoumarin antibiotic recognized by catechol transporters of E. coli.
abstract: An artificial operon was synthesized, consisting of the genes for chorismate pyruvate-lyase of E. coli and for 4-hydroxybenzoate 3-hydroxylase of Corynebacterium cyclohexanicum. This operon, directing the biosynthesis of 3,4-dihdroxybenzoate, was expressed in the heterologous expression host Streptomyces coelicolor M512, together with a modified biosynthetic gene cluster for the aminocoumarin antibiotic clorobiocin. The resulting strain produced a clorobiocin derivative containing a 3,4-dihdroxybenzoyl moiety. Its structure was confirmed by MS and NMR analysis, and it was found to be a potent inhibitor of the gyrases from Escherichia coli and Staphylococcus aureus. Bioassays against different E. coli mutants suggested that this compound is actively imported by catechol siderophore transporters in the cell envelope. This study provides an example of the structure of a natural product that can be rationally modified by synthetic biology.
authors: Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
authors: Silke
authors: S
authors: Alt
authors: Nadja
authors: N
authors: Burkard
authors: Andreas
authors: A
authors: Kulik
authors: Stephanie
authors: S
authors: Grond
authors: Lutz
authors: L
authors: Heide
mesh_terms: D015109:4-Hydroxybenzoate-3-Monooxygenase / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000494:pharmacology; D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D003352:Corynebacterium / Q000235:genetics; D027081:DNA Gyrase / Q000378:metabolism; D052978:Disk Diffusion Antimicrobial Tests; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D062385:Hydroxybenzoates / Q000737:chemistry / Q000378:metabolism*; D005810:Multigene Family; D009675:Novobiocin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000494:pharmacology; D007652:Oxo-Acid-Lyases / Q000235:genetics; D011956:Receptors, Cell Surface / Q000378:metabolism*; D048372:Streptomyces coelicolor / Q000235:genetics / Q000378:metabolism; D059005:Topoisomerase II Inhibitors
chemical_list: D000900:Anti-Bacterial Agents; D001425:Bacterial Outer Membrane Proteins; D062385:Hydroxybenzoates; D011956:Receptors, Cell Surface; D059005:Topoisomerase II Inhibitors; C559400:novobiocin 401; C035902:siderophore receptors; D009675:Novobiocin; C009091:protocatechuic acid; C006260:clorobiocin; D015109:4-Hydroxybenzoate-3-Monooxygenase; D007652:Oxo-Acid-Lyases; C075960:chorismate pyruvate lyase; D027081:DNA Gyrase