Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics

Wei Chen Chang, Yisong Guo, Chen Wang, Susan E. Butch, Amy C. Rosenzweig, Amie Kathleen Boal, Carsten Krebs, Joseph M. Bollinger, Jr.

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

The bicyclic β-lactam/2-pyrrolidine precursor to all carbapenem antibiotics is biosynthesized by attachment of a carboxymethylene unit to C5 of L-proline followed by β-lactam ring closure. Carbapenem synthase (CarC), an Fe(II) and 2-(oxo)glutarate (Fe/2OG)-dependent oxygenase, then inverts the C5 configuration. Here we report the structure of CarC in complex with its substrate and biophysical dissection of its reaction to reveal the stereoinversion mechanism. An Fe(IV)-oxo intermediate abstracts the hydrogen (H•) from C5, and tyrosine 165, a residue not visualized in the published structures of CarC lacking bound substrate, donates H• to the opposite face of the resultant radical. The reaction oxidizes the Fe(II) cofactor to Fe(III), limiting wild-type CarC to one turnover, but substitution of the H•-donating tyrosine disables stereoinversion and confers to CarC the capacity for catalytic substrate oxidation.

Original languageEnglish (US)
Pages (from-to)1140-1144
Number of pages5
JournalScience
Volume343
Issue number6175
DOIs
StatePublished - Jan 1 2014

Fingerprint

Lactams
Carbapenems
Tyrosine
Glutarates
Oxygenases
Proline
Dissection
Hydrogen
Antibiotics
Substrate
pyrrolidine

All Science Journal Classification (ASJC) codes

  • General
  • Medicine(all)
  • History and Philosophy of Science

Cite this

Chang, Wei Chen ; Guo, Yisong ; Wang, Chen ; Butch, Susan E. ; Rosenzweig, Amy C. ; Boal, Amie Kathleen ; Krebs, Carsten ; Bollinger, Jr., Joseph M. / Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics. In: Science. 2014 ; Vol. 343, No. 6175. pp. 1140-1144.
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Mechanism of the C5 stereoinversion reaction in the biosynthesis of carbapenem antibiotics. / Chang, Wei Chen; Guo, Yisong; Wang, Chen; Butch, Susan E.; Rosenzweig, Amy C.; Boal, Amie Kathleen; Krebs, Carsten; Bollinger, Jr., Joseph M.

In: Science, Vol. 343, No. 6175, 01.01.2014, p. 1140-1144.

Research output: Contribution to journalArticle

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AU - Boal, Amie Kathleen

AU - Krebs, Carsten

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