Manganese-substituted carbonic anhydrase as a new peroxidase

Krzysztof Okrasa, Romas J. Kazlauskas

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Carbonic anhydrase is a zinc metalloenzyme that catalyzes the hydration of carbon dioxide to bicarbonate. Replacing the active-site zinc with manganese yielded manganese-substituted carbonic anhydrase (CA[Mn]), which shows peroxidase activity with a bicarbonate-dependent mechanism. In the presence of bicarbonate and hydrogen peroxide, (CA[Mn]) catalyzed the efficient oxidation of o-dianisidine with kcat/KM = 1.4 × 10 6M-1S-1, which is comparable to that for horseradish peroxidase, kcat/KM = 57 × M -1 S-1. CA[Mn] also catalyzed the moderately enantioselective epoxidation of olefins to epoxides (E = 5 for p-chlorostyrene) in the presence of an amino-alcohol buffer, such as N,N-bis(2-hydroxyethyl)-2-aminoethanesulfonic acid (BES). This enantioselectivity is similar to that for natural heme-based peroxidases, but has the advantage that CA[Mn] avoids the formation of aldehyde side products. CA[Mn] degrades during the epoxidation limiting the yield of the epoxidations to <12%. Replacement of active-site residues Asn62, His64, Asn67, Gln92, or Thr200 with alanine by site-directed mutagenesis decreased the enantioselectivity demonstrating that the active site controls the enantioselectivity of the epoxidation.

Original languageEnglish (US)
Pages (from-to)1587-1596
Number of pages10
JournalChemistry - A European Journal
Issue number6
StatePublished - Feb 8 2006


  • Carbonic anhydrase
  • Enzyme catalysis
  • Epoxidation
  • Hydrogen peroxide
  • Oxidation


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