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The Applicability of Enzymes in Cellulose Ether Analysis

Adden, Roland; Melander, Claes LU ; Brinkmalm, Gunnar; Knarr, Matthias; Engelhardt, Juergen and Mischnick, Petra (2009) 50th Cellulose Symposium 2008 In Macromolecular Symposia 280. p.36-44
Abstract
Six methyl cellulose (MC) samples, one with a IDS Of 1.32 and five with a DS between 1.83 and 1.88, were degraded with five different enzymes or enzyme preparations containing endoglucanases. The main goal was to investigate whether enzymes could be used for determination of heterogeneity of the substituent distribution along the cellulose chain. To obtain information about the heterogeneity it was necessary to gather information on how the enzymes affect hydrolysis. Monomer composition and methyl distribution in the polymer chain were analyzed after total or partial random hydrolysis and appropriate derivatization by GC and MS, respectively, and used as reference data for the evaluation of the enzymatic hydrolysis. Size exclusion... (More)
Six methyl cellulose (MC) samples, one with a IDS Of 1.32 and five with a DS between 1.83 and 1.88, were degraded with five different enzymes or enzyme preparations containing endoglucanases. The main goal was to investigate whether enzymes could be used for determination of heterogeneity of the substituent distribution along the cellulose chain. To obtain information about the heterogeneity it was necessary to gather information on how the enzymes affect hydrolysis. Monomer composition and methyl distribution in the polymer chain were analyzed after total or partial random hydrolysis and appropriate derivatization by GC and MS, respectively, and used as reference data for the evaluation of the enzymatic hydrolysis. Size exclusion chromatography with multi angle light scattering and refractive index detection (SEC-MALLS/RI) was used to estimate molar mass distribution of the MCs before and after hydrolysis. Electrospray and matrix assisted laser desorption/ionization (ESI and MALDI) in combination with various MS analyzers were compared with respect to quantification of the degradation products directly and after perdeuteromethylation. Methyl group distribution in the oligomeric fractions and the average DS/DP were calculated from ESI mass spectra. With help of the reference analysis, patterns could be corrected for the unspecific contribution of end groups. By labelling and ESI-MSn, our knowledge about the tolerance of the enzyme's sub-sites with respect to the number of methyl groups could be improved. A novel standard addition method in combination with electrospray ionization ion trap mass spectrometry (ESI-IT MS) was used to determine the amount of formed oligomers. (Less)
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author
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
MALDI-MS, ESI-MS, enzyme specificity, cellulose degrading enzymes, endoglucanases, methyl cellulose, quantitative mass spectrometry
in
Macromolecular Symposia
volume
280
pages
36 - 44
publisher
John Wiley & Sons
conference name
50th Cellulose Symposium 2008
external identifiers
  • wos:000268552000006
  • scopus:67649963278
ISSN
1022-1360
1521-3900
language
English
LU publication?
yes
id
1d08e87e-8834-4414-b84f-63468cafcea4 (old id 1460095)
date added to LUP
2009-08-25 13:43:07
date last changed
2017-01-01 05:09:48
@inproceedings{1d08e87e-8834-4414-b84f-63468cafcea4,
  abstract     = {Six methyl cellulose (MC) samples, one with a IDS Of 1.32 and five with a DS between 1.83 and 1.88, were degraded with five different enzymes or enzyme preparations containing endoglucanases. The main goal was to investigate whether enzymes could be used for determination of heterogeneity of the substituent distribution along the cellulose chain. To obtain information about the heterogeneity it was necessary to gather information on how the enzymes affect hydrolysis. Monomer composition and methyl distribution in the polymer chain were analyzed after total or partial random hydrolysis and appropriate derivatization by GC and MS, respectively, and used as reference data for the evaluation of the enzymatic hydrolysis. Size exclusion chromatography with multi angle light scattering and refractive index detection (SEC-MALLS/RI) was used to estimate molar mass distribution of the MCs before and after hydrolysis. Electrospray and matrix assisted laser desorption/ionization (ESI and MALDI) in combination with various MS analyzers were compared with respect to quantification of the degradation products directly and after perdeuteromethylation. Methyl group distribution in the oligomeric fractions and the average DS/DP were calculated from ESI mass spectra. With help of the reference analysis, patterns could be corrected for the unspecific contribution of end groups. By labelling and ESI-MSn, our knowledge about the tolerance of the enzyme's sub-sites with respect to the number of methyl groups could be improved. A novel standard addition method in combination with electrospray ionization ion trap mass spectrometry (ESI-IT MS) was used to determine the amount of formed oligomers.},
  author       = {Adden, Roland and Melander, Claes and Brinkmalm, Gunnar and Knarr, Matthias and Engelhardt, Juergen and Mischnick, Petra},
  booktitle    = {Macromolecular Symposia},
  issn         = {1022-1360},
  keyword      = {MALDI-MS,ESI-MS,enzyme specificity,cellulose degrading enzymes,endoglucanases,methyl cellulose,quantitative mass spectrometry},
  language     = {eng},
  pages        = {36--44},
  publisher    = {John Wiley & Sons},
  title        = {The Applicability of Enzymes in Cellulose Ether Analysis},
  volume       = {280},
  year         = {2009},
}