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Titolo:
Stabilization of multimeric enzymes via immobilization and post-immobilization techniques
Autore:
Fernandez-Lafuente, R; Rodriguez, V; Mateo, C; Penzol, G; Hernandez-Justiz, O; Irazoqui, G; Villarino, A; Ovsejevi, K; Batista, F; Guisan, JM;
Indirizzi:
CSIC, Inst Catalisis, Dept Biocatalysis, E-28049 Madrid, Spain CSIC Madrid Spain E-28049 isis, Dept Biocatalysis, E-28049 Madrid, Spain Tech Univ Havana, Dept Biotechnol, Havana, Cuba Tech Univ Havana Havana Cuba Univ Havana, Dept Biotechnol, Havana, Cuba Univ Republ, Catedra Bioquim, Montevideo, Uruguay Univ Republ MontevideoUruguay bl, Catedra Bioquim, Montevideo, Uruguay
Titolo Testata:
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
fascicolo: 1-4, volume: 7, anno: 1999,
pagine: 181 - 189
SICI:
1381-1177(19990915)7:1-4<181:SOMEVI>2.0.ZU;2-P
Fonte:
ISI
Lingua:
ENG
Soggetto:
ALDEHYDE-AGAROSE GELS; PENICILLIN-G ACYLASE; BIFUNCTIONAL REAGENTS; ACTIVATED SUPPORTS;
Keywords:
multimeric enzymes; protein immobilization; chemical cross-linking of proteins; dextrans; stabilization of enzymes;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Physical, Chemical & Earth Sciences
Citazioni:
14
Recensione:
Indirizzi per estratti:
Indirizzo: Guisan, JM CSIC, Inst Catalisis, Dept Biocatalysis, Campus Univ Autonoma, E-28049 Madrid, Spain CSIC Campus Univ Autonoma Madrid Spain E-28049 9 Madrid, Spain
Citazione:
R. Fernandez-Lafuente et al., "Stabilization of multimeric enzymes via immobilization and post-immobilization techniques", J MOL CAT B, 7(1-4), 1999, pp. 181-189

Abstract

Controlled and directed immobilization plus post-immobilization techniquesare proposed to get full stabilization of the quaternary structure of mostmultimeric industrial enzymes. The sequential utilization of two stabilization approaches is proposed: (a) Multi-subunit immobilization: a very intense multi-subunit covalent immobilization has been achieved by performing very long immobilization processes between multimeric enzymes and porous supports composed by large internal surfaces and covered by a very dense layer of reactive groups secluded from the support surface through very short spacer arms. (b) Additional cross-linking with poly-functional macromolecules:additional chemical modification of multi-subunit immobilized derivatives with polyfunctional macromolecules promotes an additional cross-linking of all subunits of most of multimeric enzymes. A number of homo and hetero-dimeric enzymes has been stabilized by the simple application of multi-subunitimmobilization but more complex multimeric enzymes (e.g., tetrameric ones)were only fully stabilized after the sequential application of both strategies. After such stabilization of the quaternary structure these three features were observed: no subunits were desorbed from derivatives after boiling them in SDS, thermal inactivation becomes independent from enzyme concentration and derivatives became much more stable than soluble enzymes as wellas than non-stabilized derivatives. For example, thermal stability of D-amino acid oxidase from Rhodotorula gracilis was increased 7.000 fold after stabilization of its quaternary structure. (C) 1999 Elsevier Science B.V. All rights reserved.

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Documento generato il 05/07/20 alle ore 06:48:49