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Titolo:
Properties of firefly luciferase immobilized through a biotin carboxyl carrier protein domain
Autore:
Eu, JY; Andrade, J;
Indirizzi:
Univ Utah, Dept Bioengn, Salt Lake City, UT 84112 USA Univ Utah Salt LakeCity UT USA 84112 oengn, Salt Lake City, UT 84112 USA Univ Utah, Dept Mat Sci & Engn, Salt Lake City, UT 84112 USA Univ Utah Salt Lake City UT USA 84112 Engn, Salt Lake City, UT 84112 USA
Titolo Testata:
LUMINESCENCE
fascicolo: 1, volume: 16, anno: 2001,
pagine: 57 - 63
SICI:
1522-7235(200101/02)16:1<57:POFLIT>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
ATP; DETERGENTS; KINETICS; ASSAY;
Keywords:
immobilization; firefly luciferase; biotin carboxyl carrier protein (BCCP); co-enzyme A (CoA); dithiothreitol (DTT);
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
34
Recensione:
Indirizzi per estratti:
Indirizzo: Andrade, J Univ Utah, Dept Bioengn, 50 S Cent Campus Dr Rm 2480, Salt LakeCity, UT 84112 USA Univ Utah 50 S Cent Campus Dr Rm 2480 Salt Lake City UTUSA 84112
Citazione:
J.Y. Eu e J. Andrade, "Properties of firefly luciferase immobilized through a biotin carboxyl carrier protein domain", LUMINESCENC, 16(1), 2001, pp. 57-63

Abstract

A fusion protein, consisting of biotin carboxyl carrier protein (BCCP) domain from Escherichia coli and firefly luciferase (FL) from Photinus pyralis, was immobilized through the biotin-avidin interaction on 6%, cross-linkedagarose beads. Several properties of the immobilized BCCP-FL were studied. Immobilized and free enzymes showed no significant difference in thermal stability; both retained at least 91% activity after incubation at 4 degreesC and 25 degreesC for 22 h. Incubation at 37 degreesC for 22 h caused significant activity loss. K-M and k(cat) values were determined for both free and immobilized enzymes. K-M values were similar between free and immobilized enzymes; however, k(cat) of immobilized BCCP-FL, was one-third of the k(cat) of the free enzyme. 294 mu mol/L Co-enzyme A (CoA) and 44 mmol/L dithiothreitol (DTT) enhanced the total bioluminescence output. Triton X-100, Tween 20. PEG 8,000, PVP 40,000 and PVP 360,000 did not enhance the bioluminescence reaction of immobilized BCCP-FL. Copyright (C) 2001 John Wiley & Sons, Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 31/05/20 alle ore 06:05:48