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Liquid chromatography-mass spectrometry analysis of enzyme-hydrolysed carboxymethylcellulose for investigation of enzyme selectivity and substituent pattern

Cohen, Arieh LU ; Schagerlöf, Herje LU ; Nilsson, Carina LU ; Melander, Claes LU ; Tjerneld, Folke LU and Gorton, Lo LU (2004) In Journal of Chromatography A 1029(1-2). p.87-95
Abstract
A series of celloendoglucanases: Bacillus agaradhaerens Cel 5a, Humicola insolens Cel 5a, H. insolens Cel 7b, H. insolens Cel 45a, Trichoderma reesei Cel 7b, and T reesei Cel 45a were used to hydrolyse carboxymethylcellulose (CMC) and the hydrolysis products were investigated with a novel liquid chromatography-mass spectrometry (LC-MS) method. Separation was achieved using a graphitised carbon chromatographic column which allowed the use of electrospay compatible eluents. Analysis of the compounds produced during enzyme hydrolysis of CMC is used to understand enzyme selectivities and substitution pattern of CMC. Conventional high-performance anion-exchange chromatography (HPAEC)-pulsed amperometric detection (PAD), size-exclusion... (More)
A series of celloendoglucanases: Bacillus agaradhaerens Cel 5a, Humicola insolens Cel 5a, H. insolens Cel 7b, H. insolens Cel 45a, Trichoderma reesei Cel 7b, and T reesei Cel 45a were used to hydrolyse carboxymethylcellulose (CMC) and the hydrolysis products were investigated with a novel liquid chromatography-mass spectrometry (LC-MS) method. Separation was achieved using a graphitised carbon chromatographic column which allowed the use of electrospay compatible eluents. Analysis of the compounds produced during enzyme hydrolysis of CMC is used to understand enzyme selectivities and substitution pattern of CMC. Conventional high-performance anion-exchange chromatography (HPAEC)-pulsed amperometric detection (PAD), size-exclusion chromatography (SEC)-refractive index (RI) detection. and reducing end analysis are also used to analyse enzyme-hydrolysed CMC. The LC-MS method presented allows for a more detailed investigation of hydrolysis products, which facilitates characterisation of both enzymes and substrates. (C) 2004 Elsevier B.V. All rights reserved. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
cellulose, enzymes, endoglucanases, carbohydrates, carboxymethylcellulose
in
Journal of Chromatography A
volume
1029
issue
1-2
pages
87 - 95
publisher
Elsevier
external identifiers
  • pmid:15032353
  • wos:000189076000011
  • scopus:1042299051
ISSN
0021-9673
DOI
10.1016/j.chroma.2003.12.010
language
English
LU publication?
yes
id
6a5d31b4-6cec-4b8b-b73a-c03f4d5bc0d9 (old id 120872)
alternative location
http://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6TG8-4BG8V0Y-9-16&_cdi=5248&_orig=search&_coverDate=03%2F12%2F2004&_qd=1&_sk=989709998&view=c&wchp=dGLbVlz-zSkWz&_acct=C000041498&_version=1&_userid=745831&md5=d80ca1ca55b94901940d5ea0985ddd8e&ie=f.
date added to LUP
2007-06-26 13:51:25
date last changed
2017-10-22 04:34:11
@article{6a5d31b4-6cec-4b8b-b73a-c03f4d5bc0d9,
  abstract     = {A series of celloendoglucanases: Bacillus agaradhaerens Cel 5a, Humicola insolens Cel 5a, H. insolens Cel 7b, H. insolens Cel 45a, Trichoderma reesei Cel 7b, and T reesei Cel 45a were used to hydrolyse carboxymethylcellulose (CMC) and the hydrolysis products were investigated with a novel liquid chromatography-mass spectrometry (LC-MS) method. Separation was achieved using a graphitised carbon chromatographic column which allowed the use of electrospay compatible eluents. Analysis of the compounds produced during enzyme hydrolysis of CMC is used to understand enzyme selectivities and substitution pattern of CMC. Conventional high-performance anion-exchange chromatography (HPAEC)-pulsed amperometric detection (PAD), size-exclusion chromatography (SEC)-refractive index (RI) detection. and reducing end analysis are also used to analyse enzyme-hydrolysed CMC. The LC-MS method presented allows for a more detailed investigation of hydrolysis products, which facilitates characterisation of both enzymes and substrates. (C) 2004 Elsevier B.V. All rights reserved.},
  author       = {Cohen, Arieh and Schagerlöf, Herje and Nilsson, Carina and Melander, Claes and Tjerneld, Folke and Gorton, Lo},
  issn         = {0021-9673},
  keyword      = {cellulose,enzymes,endoglucanases,carbohydrates,carboxymethylcellulose},
  language     = {eng},
  number       = {1-2},
  pages        = {87--95},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Liquid chromatography-mass spectrometry analysis of enzyme-hydrolysed carboxymethylcellulose for investigation of enzyme selectivity and substituent pattern},
  url          = {http://dx.doi.org/10.1016/j.chroma.2003.12.010},
  volume       = {1029},
  year         = {2004},
}