Functionalization of Oxidases with Peroxidase Activity Creates Oxiperoxidases: A New Breed of Hybrid Enzyme Capable of Cascade Chemistry

Remko T. Winter, Tomas E. van den Berg, Dana I. Colpa, Edwin van Bloois, Marco W. Fraaije*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)
102 Downloads (Pure)

Abstract

The covalent flavoprotein alditol oxidase (AldO) from Streptomyces coelicolor A3(2) was endowed with an extra catalytic functionality by fusing it to a microperoxidase. Purification of the construct resulted in the isolation of a synthetic bifunctional enzyme that was both fully covalently flavinylated and heminylated: an oxiperoxidase. Characterization revealed that both oxidase and peroxidase functionalities were active, with the construct functioning as a single-component xylitol biosensor. In an attempt to reduce the size of the oxidaseperoxidase fusion, we replaced portions of the native AldO sequence with the bacterial cytochrome c CXXCH heme-binding motif. By mutating only three residues of the AldO protein we were able to create a functional oxidaseperoxidase hybrid.

Original languageEnglish
Pages (from-to)252-258
Number of pages7
JournalChemBioChem
Volume13
Issue number2
DOIs
Publication statusPublished - 23-Jan-2012

Keywords

  • biosensors
  • cofactors
  • enzymes
  • oxidation
  • synthetic biology
  • COELICOLOR ALDITOL OXIDASE
  • COVALENTLY BOUND FLAVIN
  • CYTOCHROME-C MATURATION
  • ESCHERICHIA-COLI
  • STREPTOMYCES-COELICOLOR
  • HORSERADISH-PEROXIDASE
  • LIGNIN DEGRADATION
  • HYDROGEN-PEROXIDE
  • ALCOHOL OXIDASE
  • MICROPEROXIDASE-11

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