In vitro and in vivo production of new aminocoumarins by a combined biochemical, genetic, and synthetic approach.
The aminocoumarin antibiotics clorobiocin, novobiocin, and coumermycin A(1) are inhibitors of bacterial gyrase. Their chemical structures contain amide bonds, formed between an aminocoumarin ring and an aromatic acyl component, which is 3-dimethylallyl-4-hydroxybenzoate in the case of novobiocin and clorobiocin. These amide bonds are formed under catalysis of the gene products of cloL, novL, and couL, respectively. We first examined the substrate specificity of the purified amide synthetases CloL, NovL, and CouL for the various analogs of the prenylated benzoate moiety. We then generated new aminocoumarin antibiotics by feeding synthetic analogs of the 3-dimethylallyl-4-hydroxybenzoate moiety to a mutant strain defective in the biosynthesis of the prenylated benzoate moiety. This resulted in the formation of 32 new aminocoumarin compounds. The structures of these compounds were elucidated using FAB-MS and (1)H-NMR spectroscopy.
Galm
Ute
U
Pharmazeutische Biologie, Pharmazeutisches Institut, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 8, 72076 Tübingen, Germany.
Dessoy
Marco Aurélio
MA
Schmidt
Jürgen
J
Wessjohann
Ludger A
LA
Heide
Lutz
L
D019732:Amide Synthases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D049933:Aminocoumarins; D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry*; D003374:Coumarins / Q000737:chemistry* / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006094:Gram-Positive Bacteria / Q000235:genetics; D006801:Humans; D009154:Mutation; D009675:Novobiocin / Q000031:analogs & derivatives* / Q000737:chemistry; D010957:Plasmids; D013379:Substrate Specificity; D015854:Up-Regulation
D049933:Aminocoumarins; D000900:Anti-Bacterial Agents; D003374:Coumarins; D009675:Novobiocin; C006260:clorobiocin; D019732:Amide Synthases; C004628:coumermycin