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Microchip immobilized enzyme reactors for hydrolysis of methyl cellulose

Melander, Claes LU ; Momcilovic, Dane LU ; Nilsson, Carina LU ; Bengtsson, Martin LU ; Schagerlöf, Herje LU ; Tjerneld, Folke LU ; Laurell, Thomas LU ; Reimann, Curt LU and Gorton, Lo LU (2005) In Analytical Chemistry 77(10). p.3284-3291
Abstract
Microchip immobilized enzyme reactors (mu IMERs) with immobilized endoglucanases were applied for the hydrolysis of methyl cellulose (MC). MCs of various molecular weights were hydrolyzed using two mu IMERs containing immobilized celloendoglucanase Cel 5A from Bacillus agaradhaerens (BaCel 5A) connected in series. Hydrolysis by the mu IMER could be confirmed from the average molar masses and molar mass distributions measured by size exclusion chromatography (SEC) with online multiangle light scattering and refractive index detection. Methylated cellooligosaccharides with degrees of polymerization (DP) between 1 and 6 formed during hydrolysis were analyzed by direct infusion electrospray ionization ion-trap mass spectrometry (ESI-ITMS).... (More)
Microchip immobilized enzyme reactors (mu IMERs) with immobilized endoglucanases were applied for the hydrolysis of methyl cellulose (MC). MCs of various molecular weights were hydrolyzed using two mu IMERs containing immobilized celloendoglucanase Cel 5A from Bacillus agaradhaerens (BaCel 5A) connected in series. Hydrolysis by the mu IMER could be confirmed from the average molar masses and molar mass distributions measured by size exclusion chromatography (SEC) with online multiangle light scattering and refractive index detection. Methylated cellooligosaccharides with degrees of polymerization (DP) between 1 and 6 formed during hydrolysis were analyzed by direct infusion electrospray ionization ion-trap mass spectrometry (ESI-ITMS). Mass spectra of mu IMER- and batch-hydrolyzed samples were compared and no significant differences were found, indicating that mu IMER hydrolysis was as efficient as conventional batch hydrolysis. A fast and automated hydrolysis with online MS detection was achieved by connecting the mu IMER to high-performance liquid chromatography and ESI-ITMS. This online separation reduced the relative intensities of interfering signals and increased the signal-to-noise ratios in MS. The mu IMER hydrolysates were also subjected to SEC interfaced with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. With this technique, oligomers with DP 3-30 could be detected. The hydrolysis by they mu-IMER was performed within 60 min, i.e. significantly faster compared with batch hydrolysis usually performed for at least 24 h. The mu IMER also allowed hydrolysis after 10 days of continuous use. The method presented in this work offers new approaches for the analysis of derivatized cellulose and provides the possibility of convenient online, fast, and more versatile analysis compared with the traditional batch method. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Analytical Chemistry
volume
77
issue
10
pages
3284 - 3291
publisher
The American Chemical Society
external identifiers
  • wos:000229206800032
  • scopus:18844378855
ISSN
1520-6882
DOI
10.1021/ac050201r
language
English
LU publication?
yes
id
e0ffc66e-8180-49db-8d11-33a33a106a69 (old id 151064)
date added to LUP
2007-06-27 11:46:42
date last changed
2017-09-10 03:38:01
@article{e0ffc66e-8180-49db-8d11-33a33a106a69,
  abstract     = {Microchip immobilized enzyme reactors (mu IMERs) with immobilized endoglucanases were applied for the hydrolysis of methyl cellulose (MC). MCs of various molecular weights were hydrolyzed using two mu IMERs containing immobilized celloendoglucanase Cel 5A from Bacillus agaradhaerens (BaCel 5A) connected in series. Hydrolysis by the mu IMER could be confirmed from the average molar masses and molar mass distributions measured by size exclusion chromatography (SEC) with online multiangle light scattering and refractive index detection. Methylated cellooligosaccharides with degrees of polymerization (DP) between 1 and 6 formed during hydrolysis were analyzed by direct infusion electrospray ionization ion-trap mass spectrometry (ESI-ITMS). Mass spectra of mu IMER- and batch-hydrolyzed samples were compared and no significant differences were found, indicating that mu IMER hydrolysis was as efficient as conventional batch hydrolysis. A fast and automated hydrolysis with online MS detection was achieved by connecting the mu IMER to high-performance liquid chromatography and ESI-ITMS. This online separation reduced the relative intensities of interfering signals and increased the signal-to-noise ratios in MS. The mu IMER hydrolysates were also subjected to SEC interfaced with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. With this technique, oligomers with DP 3-30 could be detected. The hydrolysis by they mu-IMER was performed within 60 min, i.e. significantly faster compared with batch hydrolysis usually performed for at least 24 h. The mu IMER also allowed hydrolysis after 10 days of continuous use. The method presented in this work offers new approaches for the analysis of derivatized cellulose and provides the possibility of convenient online, fast, and more versatile analysis compared with the traditional batch method.},
  author       = {Melander, Claes and Momcilovic, Dane and Nilsson, Carina and Bengtsson, Martin and Schagerlöf, Herje and Tjerneld, Folke and Laurell, Thomas and Reimann, Curt and Gorton, Lo},
  issn         = {1520-6882},
  language     = {eng},
  number       = {10},
  pages        = {3284--3291},
  publisher    = {The American Chemical Society},
  series       = {Analytical Chemistry},
  title        = {Microchip immobilized enzyme reactors for hydrolysis of methyl cellulose},
  url          = {http://dx.doi.org/10.1021/ac050201r},
  volume       = {77},
  year         = {2005},
}