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Structural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supports

2001Journal article Open access
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dc.contributor.authorBalcão, Vitor M.
dc.contributor.authorMateo, Cesar
dc.contributor.authorFernández-Lafuente, R.
dc.contributor.authorMalcata, F. Xavier
dc.contributor.authorGuisán, José M.
dc.date.accessioned2011-10-23T09:12:36Z
dc.date.available2011-10-23T09:12:36Z
dc.date.issued2001
dc.description.abstractA new protocol for the stabilization of the quaternary structure of multimeric enzymes has been attempted using as model enzyme (tetrameric) L-asparaginase from Escherichia coli. Such strategy is based upon multisubunit covalent immobilization of the enzyme onto activated supports (agarose-glutaraldehyde). Supports activated with different densities of reactive groups were used; the higher the density of groups, the higher the stabilization attained. However, because of the complexity of that enzyme, even the use of the highest densities of reactive groups was not enough to encompass all four subunits in the immobilization process. Therefore, a further chemical intersubunit cross-linking with aldehyde-dextran was pursued; these derivatives displayed a fully stabilized multimeric structure. In fact, boiling the modified enzyme derivative in the presence of sodium dodecyl sulfate and â-mercaptoethanol did not lead to release of any enzyme subunit into the medium. Such a derivative, prepared under optimal conditions, retained ca. 40% of the intrinsic activity of the free enzyme and was also functionally stabilized, with thermostabilization enhancements of ca. 3 orders of magnitude when compared with its soluble counterpart. This type of derivative may be appropriate for extracorporeal devices in the clinical treatment of acute leukemia and might thus bring about inherent advantages in that all subunits are covalently bound to the support, with a longer half-life and a virtually nil risk of subunit release into the circulating blood stream.por
dc.identifier.citationBALCÃO, Vitor M... [et al.] - Structural and Functional Stabilization of L-Asparaginase via Multisubunit Immobilization onto Highly Activated Supports. Biotechnology Progress. ISSN 1520-6033. Vol. 17, n.º 3 (2001), p. 537-542por
dc.identifier.urihttp://hdl.handle.net/10400.14/6917
dc.language.isoengpor
dc.peerreviewedyespor
dc.publisherAmerican Chemical Society and American Institute of Chemical Engineerspor
dc.relation.publisherversionThe definitive version is available at www3.interscience.wiley.compor
dc.titleStructural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supportspor
dc.typejournal article
dspace.entity.typePublication
person.familyNameBalcão
person.familyNameMalcata
person.givenNameVictor
person.givenNameFrancisco
person.identifier.ciencia-idC314-A18C-931B
person.identifier.ciencia-id1B13-38A5-35F5
person.identifier.orcid0000-0003-0772-2834
person.identifier.orcid0000-0003-3073-1659
person.identifier.ridI-2182-2012
person.identifier.scopus-author-id6603449856
person.identifier.scopus-author-id7102542478
rcaap.rightsopenAccesspor
rcaap.typearticlepor
relation.isAuthorOfPublication9b0860f7-99b1-4ded-a483-6435882272d0
relation.isAuthorOfPublicationa06c00da-0e2f-4434-8ede-4d7b22c0dfe9
relation.isAuthorOfPublication.latestForDiscoverya06c00da-0e2f-4434-8ede-4d7b22c0dfe9

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