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Mass spectrometric analysis of peptides from an immobilized lipase: focus on oxidative modifications.

Törnvall, Ulrika LU ; Melin Fürst, Camilla LU ; Hatti-Kaul, Rajni LU and Hedström, Martin LU (2009) In Rapid Communications in Mass Spectrometry 23(18). p.2959-2964
Abstract
Liquid chromatography/tandem mass spectrometry (LC/MS/MS) was used to study the primary structure of immobilized Candida antarctica lipase B (Novozym(R)435) without detaching the enzyme from the carrier. The immobilized enzyme packed in a miniature column was subjected to proteolysis and the peptides released were injected into the mass spectrometer for analysis. The set-up was utilized to determine amino acid oxidation after treatment of the biocatalyst with hydrogen peroxide. In total, sequence coverage of more than 90% was obtained, containing almost all of the amino acids sensitive to oxidation. Oxidation of methionine, tryptophan and cystine residues was observed. The flow system also allowed evaluation of the enzyme activity prior to... (More)
Liquid chromatography/tandem mass spectrometry (LC/MS/MS) was used to study the primary structure of immobilized Candida antarctica lipase B (Novozym(R)435) without detaching the enzyme from the carrier. The immobilized enzyme packed in a miniature column was subjected to proteolysis and the peptides released were injected into the mass spectrometer for analysis. The set-up was utilized to determine amino acid oxidation after treatment of the biocatalyst with hydrogen peroxide. In total, sequence coverage of more than 90% was obtained, containing almost all of the amino acids sensitive to oxidation. Oxidation of methionine, tryptophan and cystine residues was observed. The flow system also allowed evaluation of the enzyme activity prior to peptide analysis. The developed method is general and should be applicable to other immobilized enzyme systems and to different treatments. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Rapid Communications in Mass Spectrometry
volume
23
issue
18
pages
2959 - 2964
publisher
John Wiley & Sons
external identifiers
  • wos:000269708300013
  • pmid:19681098
  • scopus:69849095970
ISSN
1097-0231
DOI
10.1002/rcm.4208
language
English
LU publication?
yes
id
a7928f2c-38b0-4d93-a533-21f28b7f132e (old id 1469747)
date added to LUP
2009-09-11 09:57:41
date last changed
2017-01-01 04:35:08
@article{a7928f2c-38b0-4d93-a533-21f28b7f132e,
  abstract     = {Liquid chromatography/tandem mass spectrometry (LC/MS/MS) was used to study the primary structure of immobilized Candida antarctica lipase B (Novozym(R)435) without detaching the enzyme from the carrier. The immobilized enzyme packed in a miniature column was subjected to proteolysis and the peptides released were injected into the mass spectrometer for analysis. The set-up was utilized to determine amino acid oxidation after treatment of the biocatalyst with hydrogen peroxide. In total, sequence coverage of more than 90% was obtained, containing almost all of the amino acids sensitive to oxidation. Oxidation of methionine, tryptophan and cystine residues was observed. The flow system also allowed evaluation of the enzyme activity prior to peptide analysis. The developed method is general and should be applicable to other immobilized enzyme systems and to different treatments.},
  author       = {Törnvall, Ulrika and Melin Fürst, Camilla and Hatti-Kaul, Rajni and Hedström, Martin},
  issn         = {1097-0231},
  language     = {eng},
  number       = {18},
  pages        = {2959--2964},
  publisher    = {John Wiley & Sons},
  series       = {Rapid Communications in Mass Spectrometry},
  title        = {Mass spectrometric analysis of peptides from an immobilized lipase: focus on oxidative modifications.},
  url          = {http://dx.doi.org/10.1002/rcm.4208},
  volume       = {23},
  year         = {2009},
}