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Titolo:
Increase in conformational stability of enzymes immobilized on epoxy-activated supports by favoring additional multipoint covalent attachment
Autore:
Mateo, C; Abian, O; Fernandez-Lafuente, R; Guisan, JM;
Indirizzi:
CSIC, Inst Catalisis, Dept Biocatalisis, Madrid 28049, Spain CSIC MadridSpain 28049 atalisis, Dept Biocatalisis, Madrid 28049, Spain
Titolo Testata:
ENZYME AND MICROBIAL TECHNOLOGY
fascicolo: 7, volume: 26, anno: 2000,
pagine: 509 - 515
SICI:
0141-0229(200004)26:7<509:IICSOE>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
STABILIZATION;
Keywords:
immobilization of enzymes; stabilization of enzymes; enzyme multipoint covalent attachment; epoxy supports; Chymotrypsin; penicillin G acylase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
24
Recensione:
Indirizzi per estratti:
Indirizzo: Guisan, JM CSIC, Inst Catalisis, Dept Biocatalisis, Campus Univ Autonoma,Cantoblanco,Madrid 28049, Spain CSIC Campus Univ Autonoma,Cantoblanco Madrid Spain 28049 Spain
Citazione:
C. Mateo et al., "Increase in conformational stability of enzymes immobilized on epoxy-activated supports by favoring additional multipoint covalent attachment", ENZYME MICR, 26(7), 2000, pp. 509-515

Abstract

Epoxy supports (Eupergit C) may be very suitable to achieve the multipointcovalent attachment of proteins and enzymes, therefore, to stabilize theirthree-dimensional structure. To achieve a significant multipoint covalent attachment, the control of the experimental conditions was found to be critical. A three step immobilization/stabilization procedure is here proposed:1) the enzyme is firstly covalently immobilized under very mild experimental conditions (e.g. pH 7.0 and 20 degrees C); 2) the already immobilized enzyme is further incubated under more drastic conditions (higher pH values, longer incubation periods, etc.) to "facilitate" the formation of new covalent linkages between the immobilized enzyme molecule and the support; 3) the remaining groups of the support are blocked to stop any additional interaction between the enzyme and the support. Progressive establishment of new enzyme-support attachments was showed by the progressive irreversible covalent immobilization of several subunits of multi-subunits proteins tall non-covalent structures contained in crude extracts of different microorganism,penicillin G acylase and chymotrypsin). This multipoint covalent attachment enabled the significant thermostabilization of two relevant enzymes, (compared with the just immobilized derivatives): chymotrypsin (5-fold factor) and penicillin G acylase (18-fold factor). Bearing in mind that this stabilization was additive to that achieved by conventional immobilization, the final stabilization factor become 100-fold comparing soluble penicillin G acylase and optimal derivative. These stabilizations were observed also when the inactivations were promoted by the enzyme exposure to drastic pH valuesor the presence of cosolvents. (C) 2000 Elsevier Science Inc. All rights reserved.

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Documento generato il 07/07/20 alle ore 12:29:14