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A mass spectrometric investigation of native and oxidatively inactivated chloroperoxidase

Grey, Carl Elmon; Hedström, Martin LU and Adlercreutz, Patrick LU (2007) In ChemBioChem 8(9). p.1055-1062
Abstract
The enzyme chloroperoxidase (CPO) found in Calclariomyces fumago is able to catalyze several stereoselective oxidation reaction by using a dean oxidant, usually hydrogen peroxide (H2O2), without the need for expensive cofactor generation. CPO's lack of operational stability however, is a major limitation for its commercial use. In the present study, a capillary-LC on-line trypsin- digestion system combined with reversed-phase chromatography and mass spectrometric detection was optimized for studying the primary sequence of CPO. Samples containing native CPO, CPO treated with H2O2, and CPO oxidatively inactivated by the use of indole and H2O2 were analyzed and compared. Three oxidized peptides were found in the samples treated with H2O2.... (More)
The enzyme chloroperoxidase (CPO) found in Calclariomyces fumago is able to catalyze several stereoselective oxidation reaction by using a dean oxidant, usually hydrogen peroxide (H2O2), without the need for expensive cofactor generation. CPO's lack of operational stability however, is a major limitation for its commercial use. In the present study, a capillary-LC on-line trypsin- digestion system combined with reversed-phase chromatography and mass spectrometric detection was optimized for studying the primary sequence of CPO. Samples containing native CPO, CPO treated with H2O2, and CPO oxidatively inactivated by the use of indole and H2O2 were analyzed and compared. Three oxidized peptides were found in the samples treated with H2O2. Two additional oxidized peptides were found in the CPO samples that were completely inactivated, one of which contained an oxidized cysteine residue, Cys50, which is an essential amio acid due to its function as the axial ligand to the iron in the heme - the prosthetic group in CPO. In addition, the heme group was absent in the inactivated samples but was readily detected in other samples. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
mass spectrometry, chloroperoxidase, indole, peptides, oxidation
in
ChemBioChem
volume
8
issue
9
pages
1055 - 1062
publisher
John Wiley & Sons
external identifiers
  • wos:000247546400013
  • scopus:34547552884
ISSN
1439-4227
DOI
10.1002/cbic.200700091
language
English
LU publication?
yes
id
2cb70402-8eeb-46a9-b772-04dcd315ed2e (old id 648148)
date added to LUP
2007-12-11 10:55:20
date last changed
2017-09-10 03:46:43
@article{2cb70402-8eeb-46a9-b772-04dcd315ed2e,
  abstract     = {The enzyme chloroperoxidase (CPO) found in Calclariomyces fumago is able to catalyze several stereoselective oxidation reaction by using a dean oxidant, usually hydrogen peroxide (H2O2), without the need for expensive cofactor generation. CPO's lack of operational stability however, is a major limitation for its commercial use. In the present study, a capillary-LC on-line trypsin- digestion system combined with reversed-phase chromatography and mass spectrometric detection was optimized for studying the primary sequence of CPO. Samples containing native CPO, CPO treated with H2O2, and CPO oxidatively inactivated by the use of indole and H2O2 were analyzed and compared. Three oxidized peptides were found in the samples treated with H2O2. Two additional oxidized peptides were found in the CPO samples that were completely inactivated, one of which contained an oxidized cysteine residue, Cys50, which is an essential amio acid due to its function as the axial ligand to the iron in the heme - the prosthetic group in CPO. In addition, the heme group was absent in the inactivated samples but was readily detected in other samples.},
  author       = {Grey, Carl Elmon and Hedström, Martin and Adlercreutz, Patrick},
  issn         = {1439-4227},
  keyword      = {mass spectrometry,chloroperoxidase,indole,peptides,oxidation},
  language     = {eng},
  number       = {9},
  pages        = {1055--1062},
  publisher    = {John Wiley & Sons},
  series       = {ChemBioChem},
  title        = {A mass spectrometric investigation of native and oxidatively inactivated chloroperoxidase},
  url          = {http://dx.doi.org/10.1002/cbic.200700091},
  volume       = {8},
  year         = {2007},
}