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Endoglucanase sensitivity for substituents in methyl cellulose hydrolysis studied using MALDI-TOFMS for oligosaccharide analysis and structural analysis of enzyme active sites

Schagerlöf, Ulrika LU ; Schagerlöf, Herje LU ; Momcilovic, Dane LU ; Brinkmalm, Gunnar and Tjerneld, Folke LU (2007) In Biomacromolecules 8(8). p.2358-2365
Abstract
The properties of modified cellulose polymers, such as methylcellulose, are significantly influenced by the distribution of substituents along the polymer backbone. This distribution is difficult to determine due to the lack of suitable analytical methods. One approach is to use cellulose-degrading enzymes to gain information from the capability of the enzymes to cleave the bonds between glucose units. Endoglucanases are cellulase enzymes that can break internal glycosidic linkages and degrade low substituted regions of modified cellulose where the substituents do not interfere with the enzyme active site. In this work methyl cellulose was degraded using five endoglucanases from glycosyl hydrolase families 5 and 7 from three different... (More)
The properties of modified cellulose polymers, such as methylcellulose, are significantly influenced by the distribution of substituents along the polymer backbone. This distribution is difficult to determine due to the lack of suitable analytical methods. One approach is to use cellulose-degrading enzymes to gain information from the capability of the enzymes to cleave the bonds between glucose units. Endoglucanases are cellulase enzymes that can break internal glycosidic linkages and degrade low substituted regions of modified cellulose where the substituents do not interfere with the enzyme active site. In this work methyl cellulose was degraded using five endoglucanases from glycosyl hydrolase families 5 and 7 from three different species. The products were analyzed with reducing end analysis, chromatography (SEC-MALS-RI), and MALDI-TOFMS. The results were correlated with available determined enzyme structures and using structural alignment for unknown enzyme structures. This was performed in order to elucidate the relationship between active site structures and sensitivity for substituents on derivatized cellulose. The evaluation of endoglucanase hydrolysis of methyl cellulose showed that differences in sensitivity could be related to differences in steric hindrance of substituents in the active site, which could explain differences within family 5 and 7 enzymes, as well as the generally higher substituent tolerance for family 5 enzymes. This information is important for use of endoglucanases as tools for characterization of substituent distribution. The results are also valuable since soluble cellulose derivatives are generally used as substrates during enzyme characterization and in endoglucanase activity assays. (Less)
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published
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keywords
TRICHODERMA-REESEI, MULTIPLE SEQUENCE ALIGNMENT, DERIVATIVES, CEL5A, RESOLUTION, METHYLCELLULOSES, DEGRADATION, MOLAR-MASS, FLIGHT MASS-SPECTROMETRY, CARBOXYMETHYL CELLULOSE
in
Biomacromolecules
volume
8
issue
8
pages
2358 - 2365
publisher
The American Chemical Society (ACS)
external identifiers
  • wos:000248755000005
  • scopus:34548249734
  • pmid:17616166
ISSN
1526-4602
DOI
10.1021/bm0701200
language
English
LU publication?
yes
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The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Analytical Chemistry (S/LTH) (011001004), Biochemistry and Structural Biology (S) (000006142)
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241a2c00-b329-4679-80fc-beb2457c2371 (old id 637538)
date added to LUP
2016-04-04 12:16:58
date last changed
2020-09-02 02:58:21
@article{241a2c00-b329-4679-80fc-beb2457c2371,
  abstract     = {The properties of modified cellulose polymers, such as methylcellulose, are significantly influenced by the distribution of substituents along the polymer backbone. This distribution is difficult to determine due to the lack of suitable analytical methods. One approach is to use cellulose-degrading enzymes to gain information from the capability of the enzymes to cleave the bonds between glucose units. Endoglucanases are cellulase enzymes that can break internal glycosidic linkages and degrade low substituted regions of modified cellulose where the substituents do not interfere with the enzyme active site. In this work methyl cellulose was degraded using five endoglucanases from glycosyl hydrolase families 5 and 7 from three different species. The products were analyzed with reducing end analysis, chromatography (SEC-MALS-RI), and MALDI-TOFMS. The results were correlated with available determined enzyme structures and using structural alignment for unknown enzyme structures. This was performed in order to elucidate the relationship between active site structures and sensitivity for substituents on derivatized cellulose. The evaluation of endoglucanase hydrolysis of methyl cellulose showed that differences in sensitivity could be related to differences in steric hindrance of substituents in the active site, which could explain differences within family 5 and 7 enzymes, as well as the generally higher substituent tolerance for family 5 enzymes. This information is important for use of endoglucanases as tools for characterization of substituent distribution. The results are also valuable since soluble cellulose derivatives are generally used as substrates during enzyme characterization and in endoglucanase activity assays.},
  author       = {Schagerlöf, Ulrika and Schagerlöf, Herje and Momcilovic, Dane and Brinkmalm, Gunnar and Tjerneld, Folke},
  issn         = {1526-4602},
  language     = {eng},
  number       = {8},
  pages        = {2358--2365},
  publisher    = {The American Chemical Society (ACS)},
  series       = {Biomacromolecules},
  title        = {Endoglucanase sensitivity for substituents in methyl cellulose hydrolysis studied using MALDI-TOFMS for oligosaccharide analysis and structural analysis of enzyme active sites},
  url          = {http://dx.doi.org/10.1021/bm0701200},
  doi          = {10.1021/bm0701200},
  volume       = {8},
  year         = {2007},
}