Characterization of the aminocoumarin ligase SimL from the simocyclinone pathway and tandem incubation with NovM,P,N from the novobiocin pathway.
Simocyclinone D(8) consists of an anguicycline C-glycoside tethered by a tetraene diester linker to an aminocoumarin. Unlike the antibiotics novobiocin, clorobiocin, and coumermycin A(1), the phenolic hydroxyl group of the aminocoumarin in simocyclinone is not glycosylated with a decorated noviosyl moiety that is the pharmacophore for targeting bacterial DNA gyrase. We have expressed the Streptomyces antibioticus simocyclinone ligase SimL, purified it from Escherichia coli, and established its ATP-dependent amide bond forming activity with a variety of polyenoic acids including retinoic acid and fumagillin. We have then used the last three enzymes from the novobiocin pathway, NovM, NovP, and NovN, to convert a SimL product to a novel novobiocin analogue, in which the 3-prenyl-4-hydroxybenzoate of novobiocin is replaced with a tetraenoate moiety, to evaluate antibacterial activity.
Pacholec
Michelle
M
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
Freel Meyers
Caren L
CL
Oberthür
Markus
M
Kahne
Daniel
D
Walsh
Christopher T
CT
D019732:Amide Synthases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D049933:Aminocoumarins; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002264:Carboxylic Acids / Q000737:chemistry / Q000378:metabolism; D003374:Coumarins / Q000737:chemistry* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006027:Glycosides / Q000096:biosynthesis* / Q000737:chemistry*; D009675:Novobiocin / Q000031:analogs & derivatives* / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D013303:Streptomyces antibioticus / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D049933:Aminocoumarins; D000900:Anti-Bacterial Agents; D002264:Carboxylic Acids; D003374:Coumarins; D006027:Glycosides; C416814:simocyclinone D8; D009675:Novobiocin; C006260:clorobiocin; D019732:Amide Synthases; C004628:coumermycin