Investigation of the enzyme Bacillus agaradhaerens Cel 5a as an analytical tool in mass spectral characterisation of methylcelluloses

Arieh Cohen, Carina Nilsson, Herje Schagerlöf, Folke Tjerneld, Lo Gorton

Research output: Contribution to journalArticlepeer-review

4 Citations (SciVal)

Abstract

Two methylcelluloses (MC) were investigated by partial hydrolysis followed by direct infusion electrospray ionisation mass spectrometry (ESI-MS). Partial hydrolysis was achieved either enzymatically or by acid hydrolysis. Enzyme hydrolysis was performed using the endoglucanase Bacillus agaradhaerens Cel 5a. Three different hydrolysis buffer solutions were investigated in order to deter-mine which was most compatible with the subsequent MS analysis. ESI-MS experiments showed that when using enzymatic hydrolysis as an analytical tool for characterisation of modified cellulose one has to be watchful for the investigated polymer to have a sufficient amount of available cleavage sites in order to ensure that the original ends of the polymer do not influence the results. MS2 experiments confirmed that the detected products originated from methylcellulose and provided structural information on the investigated products. MS data proved that B. agaradhaerens Cel 5a is able to cleave beta 1 -> 4 glycosidic linkages adjacent to a permethylated glucosyl unit. MS2 data showed that methylation on the C-2 and C-6 hydroxyl groups of the glucosyl unit on the non-reducing side of the cleavage site hindered the enzyme. (c) 2006 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)16-24
JournalAnalytica Chimica Acta
Volume561
Issue number1-2
DOIs
Publication statusPublished - 2006

Bibliographical note

The information about affiliations in this record was updated in December 2015.
The record was previously connected to the following departments: Division of Occupational and Environmental Medicine (013078001), Biochemistry and Structural Biology (S) (000006142), Analytical Chemistry (S/LTH) (011001004)

Subject classification (UKÄ)

  • Analytical Chemistry

Keywords

  • Cel 5a
  • Bacillus agaradhaerens
  • ESI-MS
  • mass spectrometry
  • methylcellulose
  • MC
  • cellulase
  • modification pattern

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