Liquid chromatography combined with mass spectrometry for the investigation of endoglucanase selectivity on carboxymethyl cellulose

Jonas Enebro, Dane Momcilovic (Corresponding Author), Matti Siika-aho, Sigbritt Karlsson

Research output: Contribution to journalArticleScientificpeer-review

14 Citations (Scopus)

Abstract

Endoglucanases are useful tools in the chemical structure analysis of cellulose derivatives. However, knowledge on the endoglucanase selectivity, which is of central importance for data interpretation, is still limited. In this study, new reverse-phase liquid chromatography mass spectrometry (LC–MS) methods were developed to investigate the selectivity of the endoglucanases Cel5A, Cel7B, Cel45A, and Cel74A from the filamentous fungus Trichoderma reesei. The aim was to improve the identification of the regioisomers in the complex mixtures that are obtained after enzymatic hydrolysis. Reduction followed by per-O-methylation was performed in order to improve the separation in reverse-phase LC, increase MS sensitivity, and to facilitate structure analysis by MS/MS of O-carboxymethyl glucose and cellooligosaccharides. The cellulose selective enzymes that were investigated displayed interesting differences in enzyme selectivity on CMC substrates.
Original languageEnglish
Pages (from-to)2173-2181
Number of pages9
JournalCarbohydrate Research
Volume344
Issue number16
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Fingerprint

Carboxymethylcellulose Sodium
Cellulase
Liquid chromatography
Cellulose
Liquid Chromatography
Mass spectrometry
Mass Spectrometry
Cellulose derivatives
Trichoderma
Methylation
Enzymatic hydrolysis
Reverse-Phase Chromatography
Enzymes
Fungi
Complex Mixtures
Hydrolysis
Glucose
Substrates
endoglucanase Cel5A

Keywords

  • Carboxymethyl cellulose
  • Substituent distribution
  • Enzyme hydrolysis
  • Endoglucanase selectivity
  • Per-O-methylation
  • LC-ESI-MS/CID

Cite this

@article{c21136211e784060914700edc31045f6,
title = "Liquid chromatography combined with mass spectrometry for the investigation of endoglucanase selectivity on carboxymethyl cellulose",
abstract = "Endoglucanases are useful tools in the chemical structure analysis of cellulose derivatives. However, knowledge on the endoglucanase selectivity, which is of central importance for data interpretation, is still limited. In this study, new reverse-phase liquid chromatography mass spectrometry (LC–MS) methods were developed to investigate the selectivity of the endoglucanases Cel5A, Cel7B, Cel45A, and Cel74A from the filamentous fungus Trichoderma reesei. The aim was to improve the identification of the regioisomers in the complex mixtures that are obtained after enzymatic hydrolysis. Reduction followed by per-O-methylation was performed in order to improve the separation in reverse-phase LC, increase MS sensitivity, and to facilitate structure analysis by MS/MS of O-carboxymethyl glucose and cellooligosaccharides. The cellulose selective enzymes that were investigated displayed interesting differences in enzyme selectivity on CMC substrates.",
keywords = "Carboxymethyl cellulose, Substituent distribution, Enzyme hydrolysis, Endoglucanase selectivity, Per-O-methylation, LC-ESI-MS/CID",
author = "Jonas Enebro and Dane Momcilovic and Matti Siika-aho and Sigbritt Karlsson",
year = "2009",
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language = "English",
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pages = "2173--2181",
journal = "Carbohydrate Research",
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Liquid chromatography combined with mass spectrometry for the investigation of endoglucanase selectivity on carboxymethyl cellulose. / Enebro, Jonas; Momcilovic, Dane (Corresponding Author); Siika-aho, Matti; Karlsson, Sigbritt.

In: Carbohydrate Research, Vol. 344, No. 16, 2009, p. 2173-2181.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Liquid chromatography combined with mass spectrometry for the investigation of endoglucanase selectivity on carboxymethyl cellulose

AU - Enebro, Jonas

AU - Momcilovic, Dane

AU - Siika-aho, Matti

AU - Karlsson, Sigbritt

PY - 2009

Y1 - 2009

N2 - Endoglucanases are useful tools in the chemical structure analysis of cellulose derivatives. However, knowledge on the endoglucanase selectivity, which is of central importance for data interpretation, is still limited. In this study, new reverse-phase liquid chromatography mass spectrometry (LC–MS) methods were developed to investigate the selectivity of the endoglucanases Cel5A, Cel7B, Cel45A, and Cel74A from the filamentous fungus Trichoderma reesei. The aim was to improve the identification of the regioisomers in the complex mixtures that are obtained after enzymatic hydrolysis. Reduction followed by per-O-methylation was performed in order to improve the separation in reverse-phase LC, increase MS sensitivity, and to facilitate structure analysis by MS/MS of O-carboxymethyl glucose and cellooligosaccharides. The cellulose selective enzymes that were investigated displayed interesting differences in enzyme selectivity on CMC substrates.

AB - Endoglucanases are useful tools in the chemical structure analysis of cellulose derivatives. However, knowledge on the endoglucanase selectivity, which is of central importance for data interpretation, is still limited. In this study, new reverse-phase liquid chromatography mass spectrometry (LC–MS) methods were developed to investigate the selectivity of the endoglucanases Cel5A, Cel7B, Cel45A, and Cel74A from the filamentous fungus Trichoderma reesei. The aim was to improve the identification of the regioisomers in the complex mixtures that are obtained after enzymatic hydrolysis. Reduction followed by per-O-methylation was performed in order to improve the separation in reverse-phase LC, increase MS sensitivity, and to facilitate structure analysis by MS/MS of O-carboxymethyl glucose and cellooligosaccharides. The cellulose selective enzymes that were investigated displayed interesting differences in enzyme selectivity on CMC substrates.

KW - Carboxymethyl cellulose

KW - Substituent distribution

KW - Enzyme hydrolysis

KW - Endoglucanase selectivity

KW - Per-O-methylation

KW - LC-ESI-MS/CID

U2 - 10.1016/j.carres.2009.08.009

DO - 10.1016/j.carres.2009.08.009

M3 - Article

VL - 344

SP - 2173

EP - 2181

JO - Carbohydrate Research

JF - Carbohydrate Research

SN - 0008-6215

IS - 16

ER -