Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7

Micha Fridman, Carl J. Balibar, Tania Lupoli, Daniel Kahne, Christopher T. Walsh, Sylvie Garneau-Tsodikova

Research output: Contribution to journalArticle

Abstract

The aminocoumarin antibiotics novobiocin, clorobiocin, and coumermycin A1 are highly potent inhibitors of the bacterial type II topoisomerase DNA gyrase. The key pharmacophore of both clorobiocin and coumermycin A1, the 5-methyl-2-pyrrolylcarbonyl moiety, targets the ATP-binding site of GyrB. The 5-methyl-2-pyrrolylcarbonyl group is transferred by the acyltransferases Clo/CouN7 from the carrier proteins Clo/CouN1 to the 3′-hydroxyl of the L-noviosyl scaffold during the late steps of clorobiocin and coumermycin A1 biosynthesis. We first examined the substrate specificity of the purified thiolation domain protein CouN1 in becoming primed by the phosphopantetheinyltransferase Sfp using a variety of synthetic CoA analogues of the 5-methyl-2-pyrrolylcarbonyl moiety. The acyl-5-CouN1 thioesters were then assayed as donors to the 3′-OH group of descarbamoylnovobiocin by the acyltransferase CouN7, resulting in 21 novel variants with heterocyclic acyl groups installed on the noviosyl moiety of the aminocoumarin scaffold. Scaleup of a 5-methylthiophene derivative yielded a compound with activity against both Gram-negative and Gram-positive bacteria. The minimal inhibitory concentration found for the Gram-positive bacteria was comparable to that of novobiocin.

Original languageEnglish (US)
Pages (from-to)8462-8471
Number of pages10
JournalBiochemistry
Volume46
Issue number28
DOIs
StatePublished - Jul 17 2007

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Aminocoumarins
Novobiocin
Acyltransferases
Gram-Positive Bacteria
Anti-Bacterial Agents
Scaffolds
Bacteria
Enzymes
DNA Gyrase
Topoisomerase II Inhibitors
Biosynthesis
Coenzyme A
Substrate Specificity
Hydroxyl Radical
Carrier Proteins
Adenosine Triphosphate
Binding Sites
Derivatives
Substrates
coumermycin

ASJC Scopus subject areas

  • Biochemistry

Cite this

Fridman, M., Balibar, C. J., Lupoli, T., Kahne, D., Walsh, C. T., & Garneau-Tsodikova, S. (2007). Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7. Biochemistry, 46(28), 8462-8471. https://doi.org/10.1021/bi700433v

Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7. / Fridman, Micha; Balibar, Carl J.; Lupoli, Tania; Kahne, Daniel; Walsh, Christopher T.; Garneau-Tsodikova, Sylvie.

In: Biochemistry, Vol. 46, No. 28, 17.07.2007, p. 8462-8471.

Research output: Contribution to journalArticle

Fridman, M, Balibar, CJ, Lupoli, T, Kahne, D, Walsh, CT & Garneau-Tsodikova, S 2007, 'Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7', Biochemistry, vol. 46, no. 28, pp. 8462-8471. https://doi.org/10.1021/bi700433v
Fridman M, Balibar CJ, Lupoli T, Kahne D, Walsh CT, Garneau-Tsodikova S. Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7. Biochemistry. 2007 Jul 17;46(28):8462-8471. https://doi.org/10.1021/bi700433v
Fridman, Micha ; Balibar, Carl J. ; Lupoli, Tania ; Kahne, Daniel ; Walsh, Christopher T. ; Garneau-Tsodikova, Sylvie. / Chemoenzymatic formation of novel aminocoumarin antibiotics by the enzymes CouN1 and CouN7. In: Biochemistry. 2007 ; Vol. 46, No. 28. pp. 8462-8471.
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