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Titolo:
Purification of urease from water melon seeds for clinical diagnostic kits
Autore:
Mohamed, TM; Mohamed, MA; Mohamed, SA; Fahmy, AS;
Indirizzi:
Natl Res Ctr, Dept Mol Biol, Cairo, Egypt Natl Res Ctr Cairo EgyptNatl Res Ctr, Dept Mol Biol, Cairo, Egypt
Titolo Testata:
BIORESOURCE TECHNOLOGY
fascicolo: 3, volume: 68, anno: 1999,
pagine: 215 - 223
SICI:
0960-8524(199906)68:3<215:POUFWM>2.0.ZU;2-9
Fonte:
ISI
Lingua:
ENG
Soggetto:
BOVINE RUMEN; NICKEL; PYLORI;
Keywords:
urease; water melon; Citrullus vulgaris; purification; clinical use;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
39
Recensione:
Indirizzi per estratti:
Indirizzo: Fahmy, AS Natl Res Ctr, Dept Mol Biol, Tahrir St, Cairo, Egypt Natl Res Ctr Tahrir St Cairo Egypt ol, Tahrir St, Cairo, Egypt
Citazione:
T.M. Mohamed et al., "Purification of urease from water melon seeds for clinical diagnostic kits", BIORES TECH, 68(3), 1999, pp. 215-223

Abstract

Urease, in liquid and powder forms, with a purity meeting the requirementsof diagnostic use, were partially purified from water melon Citrullus vulgaris cv. 'Giza 1' seeds through a simple reproducible method consisting of delipidation, extraction, batch adsorption on TEAE-cellulose, filtration through Non Binding Protein Filter and lyophilization. The electrophoretic behaviour of the final preparation showed a single band for urease activity which coincided with the major protein band. To stabilize the solution form,1 mM EDTA and 10% glycerol were routinely added to the enzyme solution during purification steps. To avoid contamination with microorganisms and maintain enzyme stability, 0.1% sodium azide and 0.01 mM dithiothreitol were added to the filtered enzyme, respectively. Urease in a powder form was prepared in absence of glycerol and lyophilized in presence of 2% dextran. The final preparation had a transparent appearance with free ammonia content less than 0.01 mu g unit(-1) and was stable for 14 months at 4 degrees C. Bothliquid and powder ureases exhibited a distinct pH optimum at pH 8.0. Heat stability studies indicated that at pH 7.5 no loss of enzyme activities were recorded up to 40 degrees C for 30 min. The laboratory-prepared urea kitsgave comparable activity to that of a commercially available bioMerieux urea kit for determining blood urea nitrogen (BUN). (C) 1998 Elsevier ScienceLtd. All rights reserved.

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Documento generato il 19/09/20 alle ore 09:08:42