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Titolo:
Structural and functional stabilization of L-asparaginase via multisubunitimmobilization onto highly activated supports
Autore:
Balcao, VM; Mateo, C; Fernandez-Lafuente, R; Malcata, FX; Guisan, JM;
Indirizzi:
Univ Catolica Portuguesa, Escola Super Biotecnol, P-4200072 Oporto, Portugal Univ Catolica Portuguesa Oporto Portugal P-4200072 0072 Oporto, Portugal Univ Fernando Pessoa, P-4249004 Porto, Portugal Univ Fernando Pessoa Porto Portugal P-4249004 P-4249004 Porto, Portugal Univ Autonoma Madrid, Inst Catalisis & Petroquim, CSIC, E-28049 Madrid, Spain Univ Autonoma Madrid Madrid Spain E-28049 m, CSIC, E-28049 Madrid, Spain
Titolo Testata:
BIOTECHNOLOGY PROGRESS
fascicolo: 3, volume: 17, anno: 2001,
pagine: 537 - 542
SICI:
8756-7938(200105/06)17:3<537:SAFSOL>2.0.ZU;2-H
Fonte:
ISI
Lingua:
ENG
Soggetto:
POLY(ETHYLENE GLYCOL)-ALBUMIN HYDROGEL; SALT-INDUCED IMMOBILIZATION; CRYSTAL-STRUCTURE; IN-VITRO; ENZYMES; DERIVATIVES; PERFORMANCE; PROTEINS; MACROMOLECULES; PURIFICATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
36
Recensione:
Indirizzi per estratti:
Indirizzo: Malcata, FX Univ Catolica Portuguesa, Escola Super Biotecnol, Rua Dr Antonio Bernardino de Almeida, P-4200072 Oporto, Portugal Univ Catolica Portuguesa Rua Dr Antonio Bernardino de Almeida Oporto Portugal P-4200072
Citazione:
V.M. Balcao et al., "Structural and functional stabilization of L-asparaginase via multisubunitimmobilization onto highly activated supports", BIOTECH PR, 17(3), 2001, pp. 537-542

Abstract

A 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 foursubunits in the immobilization process. Therefore, a further chemical intersubunit cross-linking with aldehyde-dextran was pursued; these derivativesdisplayed a fully stabilized multimeric structure. In fact, boiling the modified enzyme derivative in the presence of sodium dodecyl sulfate and P-mercaptoethanol did not lead to release of any enzyme subunit into the medium. Such a derivative, prepared under optimal conditions, retained ca. 40% ofthe 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 virtuallynil risk of subunit release into the circulating blood stream.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 19:28:15