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Insight into stability of CotA laccase from the spore coat of Bacillus subtilis

2007-12Journal article Open access
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dc.contributor.authorEP, Melo
dc.contributor.authorFernandes, A. T.
dc.contributor.authorDurao, P.
dc.contributor.authorMartins, L. O.
dc.date.accessioned2018-12-07T14:52:50Z
dc.date.available2018-12-07T14:52:50Z
dc.date.issued2007-12
dc.description.abstractThe axial ligand of the catalytic mononuclear T1 copper site (Met(502)) of the CotA laccase was replaced by a leucine or phenylalanine residue to increase the redox potential of the enzyme. These mutations led to an increase in the redox potential by approx. 100 mV relative to the wild-type enzyme but the catalytic constant k(cat) in the mutant enzymes was severely compromised. This decrease in the catalytic efficiency was unexpected as the X-ray analysis of mutants has shown that replacement of methionine ligand did not lead to major structural changes in the geometry of the T1 Centre or in the overall fold of the enzyme. However, the mutations have a profound impact on the thermodynamic stability of the enzyme. The fold of the enzyme has become unstable especially with the introduction of the larger phenylalanine residue and this instability should be related to the decrease in the catalytic efficiency. The instability of the fold for the mutant proteins resulted in the accumulation of an intermediate state, partly unfolded, in-between native and unfolded states. Quenching of tryptophan fluorescence by acrylamide has further revealed that the intermediate state is partly unfolded.
dc.description.versioninfo:eu-repo/semantics/publishedVersion
dc.identifier.doi10.1042/BST0351579
dc.identifier.issn0300-5127
dc.identifier.urihttp://hdl.handle.net/10400.1/11233
dc.language.isoeng
dc.peerreviewedyes
dc.publisherPortland Press
dc.relationNano-Engineering of Bacterial Laccases
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectTryptophan residues
dc.subjectEndospore coat
dc.subjectFluorescence
dc.subjectComponent
dc.subjectProteins
dc.titleInsight into stability of CotA laccase from the spore coat of Bacillus subtilis
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleNano-Engineering of Bacterial Laccases
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/POCI/POCI%2FBIO%2F57083%2F2004/PT
oaire.citation.endPage1582
oaire.citation.startPage1579
oaire.citation.titleBiochemical Society Transactions
oaire.citation.volume35
oaire.fundingStreamPOCI
person.familyNamePinho Melo
person.givenNameEduardo
person.identifier1443188
person.identifier.ciencia-id3C1C-C10C-1510
person.identifier.orcid0000-0002-0974-8977
person.identifier.scopus-author-id35566177900
project.funder.identifierhttp://doi.org/10.13039/501100001871
project.funder.nameFundação para a Ciência e a Tecnologia
rcaap.rightsopenAccess
rcaap.typearticle
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relation.isAuthorOfPublication.latestForDiscovery5fa1895f-5577-4652-961a-886ec9bf41b1
relation.isProjectOfPublicationc9b56ae8-3c8b-473a-ad51-e9b71097c8d4
relation.isProjectOfPublication.latestForDiscoveryc9b56ae8-3c8b-473a-ad51-e9b71097c8d4

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