The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium
peer reviewed The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three dis...
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Portland Press Ltd
2007
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Online Access: | https://orbi.uliege.be/handle/2268/6000 https://orbi.uliege.be/bitstream/2268/6000/1/Sonan-Cellulase.pdf https://doi.org/10.1042/BJ20070640 |
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ftorbi:oai:orbi.ulg.ac.be:2268/6000 2024-04-21T07:52:08+00:00 The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles 2007-10-15 https://orbi.uliege.be/handle/2268/6000 https://orbi.uliege.be/bitstream/2268/6000/1/Sonan-Cellulase.pdf https://doi.org/10.1042/BJ20070640 en eng Portland Press Ltd urn:issn:0264-6021 urn:issn:1470-8728 https://orbi.uliege.be/handle/2268/6000 info:hdl:2268/6000 https://orbi.uliege.be/bitstream/2268/6000/1/Sonan-Cellulase.pdf doi:10.1042/BJ20070640 scopus-id:2-s2.0-35048812937 info:pmid:17635108 open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess Biochemical Journal, 407 (Part 2), 293-302 (2007-10-15) cellulase linker Pseudoalteromonas haloplanktis psychrophile small-angle X-ray scattering (SAXS) Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire journal article http://purl.org/coar/resource_type/c_6501 info:eu-repo/semantics/article peer reviewed 2007 ftorbi https://doi.org/10.1042/BJ20070640 2024-03-27T14:54:36Z peer reviewed The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three disulfide bridges. To evaluate the possible role of this region in cold adaptation, the LR was sequentially shortened by protein engineering, successively deleting one and two loops of this module, whereas the last disulfide bridge was also suppressed by replacing the last two cysteine residue by two alanine residues. The kinetic and thermodynamic properties of the mutants were compared with those of the full-length enzyme, and also with those of the cold-adapted CM alone and with those of the homologous mesophilic enzyme, Cel5A, from Erwinia chrysanthemi. The thermostability of the mutated enzymes as well as their relative flexibility were evaluated by differential scanning calorimetry and fluorescence quenching respectively. The topology of the structure of the shortest mutant was determined by SAXS (small-angle X-ray scattering). The data indicate that the sequential shortening of the LR induces a regular decrease of the specific activity towards macromolecular substrates, reduces the relative flexibility and concomitantly increases the thermostability of the shortened enzymes. This demonstrates that the long LR of the full-length enzyme favours the catalytic efficiency at low and moderate temperatures by rendering the structure not only less compact, but also less stable, and plays a crucial role in the adaptation to cold of this cellulolytic enzyme. Article in Journal/Newspaper Antarc* Antarctic University of Liège: ORBi (Open Repository and Bibliography) Biochemical Journal 407 2 293 302 |
institution |
Open Polar |
collection |
University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
English |
topic |
cellulase linker Pseudoalteromonas haloplanktis psychrophile small-angle X-ray scattering (SAXS) Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
spellingShingle |
cellulase linker Pseudoalteromonas haloplanktis psychrophile small-angle X-ray scattering (SAXS) Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
topic_facet |
cellulase linker Pseudoalteromonas haloplanktis psychrophile small-angle X-ray scattering (SAXS) Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
description |
peer reviewed The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three disulfide bridges. To evaluate the possible role of this region in cold adaptation, the LR was sequentially shortened by protein engineering, successively deleting one and two loops of this module, whereas the last disulfide bridge was also suppressed by replacing the last two cysteine residue by two alanine residues. The kinetic and thermodynamic properties of the mutants were compared with those of the full-length enzyme, and also with those of the cold-adapted CM alone and with those of the homologous mesophilic enzyme, Cel5A, from Erwinia chrysanthemi. The thermostability of the mutated enzymes as well as their relative flexibility were evaluated by differential scanning calorimetry and fluorescence quenching respectively. The topology of the structure of the shortest mutant was determined by SAXS (small-angle X-ray scattering). The data indicate that the sequential shortening of the LR induces a regular decrease of the specific activity towards macromolecular substrates, reduces the relative flexibility and concomitantly increases the thermostability of the shortened enzymes. This demonstrates that the long LR of the full-length enzyme favours the catalytic efficiency at low and moderate temperatures by rendering the structure not only less compact, but also less stable, and plays a crucial role in the adaptation to cold of this cellulolytic enzyme. |
format |
Article in Journal/Newspaper |
author |
Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles |
author_facet |
Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles |
author_sort |
Sonan, Guillaume K |
title |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
title_short |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
title_full |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
title_fullStr |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
title_full_unstemmed |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium |
title_sort |
linker region plays a key role in the adaptation to cold of the cellulase from an antarctic bacterium |
publisher |
Portland Press Ltd |
publishDate |
2007 |
url |
https://orbi.uliege.be/handle/2268/6000 https://orbi.uliege.be/bitstream/2268/6000/1/Sonan-Cellulase.pdf https://doi.org/10.1042/BJ20070640 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Biochemical Journal, 407 (Part 2), 293-302 (2007-10-15) |
op_relation |
urn:issn:0264-6021 urn:issn:1470-8728 https://orbi.uliege.be/handle/2268/6000 info:hdl:2268/6000 https://orbi.uliege.be/bitstream/2268/6000/1/Sonan-Cellulase.pdf doi:10.1042/BJ20070640 scopus-id:2-s2.0-35048812937 info:pmid:17635108 |
op_rights |
open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1042/BJ20070640 |
container_title |
Biochemical Journal |
container_volume |
407 |
container_issue |
2 |
container_start_page |
293 |
op_container_end_page |
302 |
_version_ |
1796935386883162112 |