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 language | English |
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Pages (from-to) | 252-258 |
Number of pages | 7 |
Journal | ChemBioChem |
Volume | 13 |
Issue number | 2 |
DOIs | |
Publication status | Published - 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