Cold adaptation of a psychrophilic chitinase: a mutagenesis study
peer reviewed The gene encoding chitinase ArChiB from the Antarctic Arthrobacter sp. TAD20 has been expressed in Escherichia coli and the recombinant enzyme purified to homogeneity. In an effort to engineer cold-adapted biocatalysts through rational redesign to operate at elevated temperatures, we p...
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ftorbi:oai:orbi.ulg.ac.be:2268/15821 2024-04-21T07:49:03+00:00 Cold adaptation of a psychrophilic chitinase: a mutagenesis study Mavromatis, K. Feller, Georges Kokkinidis, M. Bouriotis, V. 2003-07 https://orbi.uliege.be/handle/2268/15821 https://orbi.uliege.be/bitstream/2268/15821/1/ProtEng_Mavromatis_2003.pdf https://doi.org/10.1093/protein/gzg069 en eng Oxford Univ Press urn:issn:0269-2139 urn:issn:1460-213X https://orbi.uliege.be/handle/2268/15821 info:hdl:2268/15821 https://orbi.uliege.be/bitstream/2268/15821/1/ProtEng_Mavromatis_2003.pdf doi:10.1093/protein/gzg069 scopus-id:2-s2.0-0042379946 info:pmid:12915727 open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess Protein Engineering, 16 (7), 497-503 (2003-07) chitinases cold adaptation mutagenesis psychrophilic enzyme thermostability 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 2003 ftorbi https://doi.org/10.1093/protein/gzg069 2024-03-27T14:52:24Z peer reviewed The gene encoding chitinase ArChiB from the Antarctic Arthrobacter sp. TAD20 has been expressed in Escherichia coli and the recombinant enzyme purified to homogeneity. In an effort to engineer cold-adapted biocatalysts through rational redesign to operate at elevated temperatures, we performed several mutations aiming to increase the rigidity of the molecular edifice of the selected psychrophilic chitinase. The mutations were designed on the basis of a homology-based three-dimensional model of the enzyme, and included an attempt to introduce a salt bridge (mutant N198K) and replacements of selected Gly residues by either Pro (mutants G93P, G254P) or Gln (G406Q). Mutant N198K resulted in a more stable protein (DeltaT(m)=0.6degreesC). Mutant G93P exhibited a DeltaT(m) of 1.2degreesC, while mutants G254P and G406Q exhibited decreased stability. We conclude that the effect of mutating Gly residues on enzyme stability is rather complex and can only be understood in the context of the structural environment. Kinetic and spectroscopic analysis of these enzyme variants revealed that the kinetic parameters k(cat) and K-m have been significantly modified. Article in Journal/Newspaper Antarc* Antarctic University of Liège: ORBi (Open Repository and Bibliography) Protein Engineering Design and Selection 16 7 497 503 |
institution |
Open Polar |
collection |
University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
English |
topic |
chitinases cold adaptation mutagenesis psychrophilic enzyme thermostability Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
spellingShingle |
chitinases cold adaptation mutagenesis psychrophilic enzyme thermostability Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire Mavromatis, K. Feller, Georges Kokkinidis, M. Bouriotis, V. Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
topic_facet |
chitinases cold adaptation mutagenesis psychrophilic enzyme thermostability Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
description |
peer reviewed The gene encoding chitinase ArChiB from the Antarctic Arthrobacter sp. TAD20 has been expressed in Escherichia coli and the recombinant enzyme purified to homogeneity. In an effort to engineer cold-adapted biocatalysts through rational redesign to operate at elevated temperatures, we performed several mutations aiming to increase the rigidity of the molecular edifice of the selected psychrophilic chitinase. The mutations were designed on the basis of a homology-based three-dimensional model of the enzyme, and included an attempt to introduce a salt bridge (mutant N198K) and replacements of selected Gly residues by either Pro (mutants G93P, G254P) or Gln (G406Q). Mutant N198K resulted in a more stable protein (DeltaT(m)=0.6degreesC). Mutant G93P exhibited a DeltaT(m) of 1.2degreesC, while mutants G254P and G406Q exhibited decreased stability. We conclude that the effect of mutating Gly residues on enzyme stability is rather complex and can only be understood in the context of the structural environment. Kinetic and spectroscopic analysis of these enzyme variants revealed that the kinetic parameters k(cat) and K-m have been significantly modified. |
format |
Article in Journal/Newspaper |
author |
Mavromatis, K. Feller, Georges Kokkinidis, M. Bouriotis, V. |
author_facet |
Mavromatis, K. Feller, Georges Kokkinidis, M. Bouriotis, V. |
author_sort |
Mavromatis, K. |
title |
Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
title_short |
Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
title_full |
Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
title_fullStr |
Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
title_full_unstemmed |
Cold adaptation of a psychrophilic chitinase: a mutagenesis study |
title_sort |
cold adaptation of a psychrophilic chitinase: a mutagenesis study |
publisher |
Oxford Univ Press |
publishDate |
2003 |
url |
https://orbi.uliege.be/handle/2268/15821 https://orbi.uliege.be/bitstream/2268/15821/1/ProtEng_Mavromatis_2003.pdf https://doi.org/10.1093/protein/gzg069 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Protein Engineering, 16 (7), 497-503 (2003-07) |
op_relation |
urn:issn:0269-2139 urn:issn:1460-213X https://orbi.uliege.be/handle/2268/15821 info:hdl:2268/15821 https://orbi.uliege.be/bitstream/2268/15821/1/ProtEng_Mavromatis_2003.pdf doi:10.1093/protein/gzg069 scopus-id:2-s2.0-0042379946 info:pmid:12915727 |
op_rights |
open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1093/protein/gzg069 |
container_title |
Protein Engineering Design and Selection |
container_volume |
16 |
container_issue |
7 |
container_start_page |
497 |
op_container_end_page |
503 |
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1796952946547621888 |