Characterization of the aminocoumarin ligase SimL from the simocyclinone pathway and tandem incubation with NovM,P,N from the novobiocin pathway

Michelle Pacholec, Caren L.Freel Meyers, Markus Oberthür, Daniel Kahne, Christopher T. Walsh

Research output: Contribution to journalArticle

Abstract

Simocyclinone D8 consists of an anguicycline C-glycoside tethered by a tetraene diester linker to an aminocoumarin. Unlike the antibiotics novobiocin, clorobiocin, and coumermycin A1, 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.

Original languageEnglish (US)
Pages (from-to)4949-4956
Number of pages8
JournalBiochemistry
Volume44
Issue number12
DOIs
StatePublished - Mar 29 2005
Externally publishedYes

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ASJC Scopus subject areas

  • Biochemistry

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