Cited 4 times in
Identification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis
DC Field | Value | Language |
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dc.contributor.author | 윤호근 | - |
dc.date.accessioned | 2016-02-19T10:54:19Z | - |
dc.date.available | 2016-02-19T10:54:19Z | - |
dc.date.issued | 2001 | - |
dc.identifier.issn | 0175-7598 | - |
dc.identifier.uri | https://ir.ymlib.yonsei.ac.kr/handle/22282913/141961 | - |
dc.description.abstract | The functional importance of a conserved region in a novel chitosanase from Bacillus sp. CK4 was investigated. Each of the three carboxylic amino acid residues (Glu-50, Glu-62, and Asp-66) was changed to Asp and Gln or Asn and Glu by site-directed mutagenesis, respectively. The Asp-66→Asn and Asp-66→Glu mutation remarkably decreased kinetic parameters such as V max and k cat to approximately 1/1,000 those of the wild-type enzyme, indicating that the Asp-66 residue was essential for catalysis. The thermostable chitosanase contains three Cys residues at positions 49, 72, and 211. The Cys-49→Ser/Tyr and Cys-72→Ser/Tyr mutant enzymes were as stable to thermal inactivation and denaturating agents as the wild-type enzyme. However, the half-life of the Cys-211→Ser/Tyr mutant enzyme was less than 10 min at 80 °C, while that of the wild-type enzyme was about 90 min. Moreover, the residual activity of Cys-211→Ser/Tyr enzyme was substantially decreased by 8 M urea; and it lost all catalytic activity in 40% ethanol. These results show that the substitution of Cys with any amino acid residues at position 211 seems to affect the conformational stability of the chitosanase. | - |
dc.description.statementOfResponsibility | open | - |
dc.relation.isPartOf | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | - |
dc.rights | CC BY-NC-ND 2.0 KR | - |
dc.rights.uri | https://creativecommons.org/licenses/by-nc-nd/2.0/kr/ | - |
dc.subject.MESH | Amino Acid Sequence | - |
dc.subject.MESH | Catalysis | - |
dc.subject.MESH | Circular Dichroism | - |
dc.subject.MESH | Enzyme Stability | - |
dc.subject.MESH | Glycoside Hydrolases/chemistry* | - |
dc.subject.MESH | Glycoside Hydrolases/metabolism | - |
dc.subject.MESH | Hot Temperature | - |
dc.subject.MESH | Kinetics | - |
dc.subject.MESH | Molecular Sequence Data | - |
dc.subject.MESH | Mutagenesis, Site-Directed | - |
dc.subject.MESH | Protein Conformation | - |
dc.title | Identification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis | - |
dc.type | Article | - |
dc.contributor.college | College of Medicine (의과대학) | - |
dc.contributor.department | Dept. of Biochemistry & Molecular Biology (생화학,분자생물학) | - |
dc.contributor.googleauthor | H.-G. Yoon | - |
dc.contributor.googleauthor | H.-Y. Kim | - |
dc.contributor.googleauthor | Y.-H. Lim | - |
dc.contributor.googleauthor | H.-K. Kim | - |
dc.contributor.googleauthor | D.-H. Shin | - |
dc.contributor.googleauthor | B.-S. Hong | - |
dc.contributor.googleauthor | H.-Y. Cho | - |
dc.identifier.doi | 10.1007/s002530000571 | - |
dc.admin.author | false | - |
dc.admin.mapping | false | - |
dc.contributor.localId | A02625 | - |
dc.relation.journalcode | J00201 | - |
dc.identifier.eissn | 1432-0614 | - |
dc.identifier.pmid | 11499927 | - |
dc.identifier.url | http://link.springer.com/article/10.1007/s002530000571 | - |
dc.subject.keyword | Enzyme | - |
dc.subject.keyword | Catalytic Activity | - |
dc.subject.keyword | Bacillus | - |
dc.subject.keyword | Amino Acid Residue | - |
dc.subject.keyword | Residual Activity | - |
dc.contributor.alternativeName | Yoon, Ho Geun | - |
dc.contributor.affiliatedAuthor | Yoon, Ho Geun | - |
dc.rights.accessRights | not free | - |
dc.citation.volume | 56 | - |
dc.citation.number | 1~2 | - |
dc.citation.startPage | 173 | - |
dc.citation.endPage | 180 | - |
dc.identifier.bibliographicCitation | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, Vol.56(1~2) : 173-180, 2001 | - |
dc.identifier.rimsid | 35883 | - |
dc.type.rims | ART | - |
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