Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium
peer reviewed The gene archb encoding for the cell-bound chitobiase from the Antarctic Gram-positive bacterium Arthrobacter sp. TAD20 was cloned and expressed in Escherichia coli in a soluble form. The mature chitobiase ArChb possesses four functionally independent domains: a catalytic domain stabil...
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Online Access: | https://orbi.uliege.be/handle/2268/16035 https://doi.org/10.1006/jmbi.2001.4774 |
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ftorbi:oai:orbi.ulg.ac.be:2268/16035 2024-10-13T14:03:04+00:00 Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium Lonhienne, T. Zoidakis, J. Vorgias, C. E. Feller, Georges Gerday, Charles Bouriotis, V. 2001-07-06 https://orbi.uliege.be/handle/2268/16035 https://doi.org/10.1006/jmbi.2001.4774 en eng Elsevier urn:issn:0022-2836 urn:issn:1089-8638 https://orbi.uliege.be/handle/2268/16035 info:hdl:2268/16035 doi:10.1006/jmbi.2001.4774 info:pmid:11428890 restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess Journal of Molecular Biology, 310 (2), 291-7 (2001-07-06) 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 2001 ftorbi https://doi.org/10.1006/jmbi.2001.4774 2024-09-27T07:01:36Z peer reviewed The gene archb encoding for the cell-bound chitobiase from the Antarctic Gram-positive bacterium Arthrobacter sp. TAD20 was cloned and expressed in Escherichia coli in a soluble form. The mature chitobiase ArChb possesses four functionally independent domains: a catalytic domain stabilized by Ca(2+), a galactose-binding domain and an immunoglobulin-like domain followed by a cell-wall anchorage signal, typical of cell-surface proteins from Gram-positive bacteria. Binding of saccharides was analyzed by differential scanning calorimetry, allowing to distinguish unequivocally the catalytic domain from the galactose-binding domain and to study binding specificities. The results suggest that ArChb could play a role in bacterium attachment to natural hosts. Kinetic parameters of ArChb demonstrate perfect adaptation to catalysis at low temperatures, as shown by a low activation energy associated with unusually low K(m) and high k(cat) values. Thermodependence of these parameters indicates that discrete amino acid substitutions in the catalytic center have optimized the thermodynamic properties of weak interactions involved in substrate binding at low temperatures. Microcalorimetry also reveals that heat-lability, a general trait of psychrophilic enzymes, only affects the active site domain of ArChb. Article in Journal/Newspaper Antarc* Antarctic University of Liège: ORBi (Open Repository and Bibliography) Anchorage Antarctic The Antarctic Journal of Molecular Biology 310 2 291 297 |
institution |
Open Polar |
collection |
University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
English |
topic |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
spellingShingle |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire Lonhienne, T. Zoidakis, J. Vorgias, C. E. Feller, Georges Gerday, Charles Bouriotis, V. Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
topic_facet |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
description |
peer reviewed The gene archb encoding for the cell-bound chitobiase from the Antarctic Gram-positive bacterium Arthrobacter sp. TAD20 was cloned and expressed in Escherichia coli in a soluble form. The mature chitobiase ArChb possesses four functionally independent domains: a catalytic domain stabilized by Ca(2+), a galactose-binding domain and an immunoglobulin-like domain followed by a cell-wall anchorage signal, typical of cell-surface proteins from Gram-positive bacteria. Binding of saccharides was analyzed by differential scanning calorimetry, allowing to distinguish unequivocally the catalytic domain from the galactose-binding domain and to study binding specificities. The results suggest that ArChb could play a role in bacterium attachment to natural hosts. Kinetic parameters of ArChb demonstrate perfect adaptation to catalysis at low temperatures, as shown by a low activation energy associated with unusually low K(m) and high k(cat) values. Thermodependence of these parameters indicates that discrete amino acid substitutions in the catalytic center have optimized the thermodynamic properties of weak interactions involved in substrate binding at low temperatures. Microcalorimetry also reveals that heat-lability, a general trait of psychrophilic enzymes, only affects the active site domain of ArChb. |
format |
Article in Journal/Newspaper |
author |
Lonhienne, T. Zoidakis, J. Vorgias, C. E. Feller, Georges Gerday, Charles Bouriotis, V. |
author_facet |
Lonhienne, T. Zoidakis, J. Vorgias, C. E. Feller, Georges Gerday, Charles Bouriotis, V. |
author_sort |
Lonhienne, T. |
title |
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
title_short |
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
title_full |
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
title_fullStr |
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
title_full_unstemmed |
Modular Structure, Local Flexibility and Cold-Activity of a Novel Chitobiase from a Psychrophilic Antarctic Bacterium |
title_sort |
modular structure, local flexibility and cold-activity of a novel chitobiase from a psychrophilic antarctic bacterium |
publisher |
Elsevier |
publishDate |
2001 |
url |
https://orbi.uliege.be/handle/2268/16035 https://doi.org/10.1006/jmbi.2001.4774 |
geographic |
Anchorage Antarctic The Antarctic |
geographic_facet |
Anchorage Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Journal of Molecular Biology, 310 (2), 291-7 (2001-07-06) |
op_relation |
urn:issn:0022-2836 urn:issn:1089-8638 https://orbi.uliege.be/handle/2268/16035 info:hdl:2268/16035 doi:10.1006/jmbi.2001.4774 info:pmid:11428890 |
op_rights |
restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess |
op_doi |
https://doi.org/10.1006/jmbi.2001.4774 |
container_title |
Journal of Molecular Biology |
container_volume |
310 |
container_issue |
2 |
container_start_page |
291 |
op_container_end_page |
297 |
_version_ |
1812819503399043072 |