0 294

Cited 4 times in

Identification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis

DC FieldValueLanguage
dc.contributor.author윤호근-
dc.date.accessioned2016-02-19T10:54:19Z-
dc.date.available2016-02-19T10:54:19Z-
dc.date.issued2001-
dc.identifier.issn0175-7598-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/141961-
dc.description.abstractThe 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.statementOfResponsibilityopen-
dc.relation.isPartOfAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.subject.MESHAmino Acid Sequence-
dc.subject.MESHCatalysis-
dc.subject.MESHCircular Dichroism-
dc.subject.MESHEnzyme Stability-
dc.subject.MESHGlycoside Hydrolases/chemistry*-
dc.subject.MESHGlycoside Hydrolases/metabolism-
dc.subject.MESHHot Temperature-
dc.subject.MESHKinetics-
dc.subject.MESHMolecular Sequence Data-
dc.subject.MESHMutagenesis, Site-Directed-
dc.subject.MESHProtein Conformation-
dc.titleIdentification of essential amino acid residues for catalytic activity and thermostability of novel chitosanase by site-directed mutagenesis-
dc.typeArticle-
dc.contributor.collegeCollege of Medicine (의과대학)-
dc.contributor.departmentDept. of Biochemistry & Molecular Biology (생화학,분자생물학)-
dc.contributor.googleauthorH.-G. Yoon-
dc.contributor.googleauthorH.-Y. Kim-
dc.contributor.googleauthorY.-H. Lim-
dc.contributor.googleauthorH.-K. Kim-
dc.contributor.googleauthorD.-H. Shin-
dc.contributor.googleauthorB.-S. Hong-
dc.contributor.googleauthorH.-Y. Cho-
dc.identifier.doi10.1007/s002530000571-
dc.admin.authorfalse-
dc.admin.mappingfalse-
dc.contributor.localIdA02625-
dc.relation.journalcodeJ00201-
dc.identifier.eissn1432-0614-
dc.identifier.pmid11499927-
dc.identifier.urlhttp://link.springer.com/article/10.1007/s002530000571-
dc.subject.keywordEnzyme-
dc.subject.keywordCatalytic Activity-
dc.subject.keywordBacillus-
dc.subject.keywordAmino Acid Residue-
dc.subject.keywordResidual Activity-
dc.contributor.alternativeNameYoon, Ho Geun-
dc.contributor.affiliatedAuthorYoon, Ho Geun-
dc.rights.accessRightsnot free-
dc.citation.volume56-
dc.citation.number1~2-
dc.citation.startPage173-
dc.citation.endPage180-
dc.identifier.bibliographicCitationAPPLIED MICROBIOLOGY AND BIOTECHNOLOGY, Vol.56(1~2) : 173-180, 2001-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Biochemistry and Molecular Biology (생화학-분자생물학교실) > 1. Journal Papers

qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.