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Titolo:
Rapid process for purification of an extracellular beta-xylosidase by aqueous two-phase extraction
Autore:
Pan, IH; Li, YK;
Indirizzi:
Natl Chiao Tung Univ, Dept Appl Chem, Hsinchu 30050, Taiwan Natl Chiao Tung Univ Hsinchu Taiwan 30050 pl Chem, Hsinchu 30050, Taiwan
Titolo Testata:
JOURNAL OF CHROMATOGRAPHY B
fascicolo: 1, volume: 754, anno: 2001,
pagine: 179 - 184
SICI:
1387-2273(20010415)754:1<179:RPFPOA>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
TRICHODERMA-KONINGII G-39; AFFINITY PRECIPITATION; LACTATE-DEHYDROGENASE; 2-PHASE SYSTEMS; PROTEINS;
Keywords:
aqueous two-phase extraction; enzymes; beta-xylosidase;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
26
Recensione:
Indirizzi per estratti:
Indirizzo: Li, YK Natl Chiao Tung Univ, Dept Appl Chem, 1001 Ta Hseuh Rd, Hsinchu 30050, Taiwan Natl Chiao Tung Univ 1001 Ta Hseuh Rd Hsinchu Taiwan 30050 Taiwan
Citazione:
I.H. Pan e Y.K. Li, "Rapid process for purification of an extracellular beta-xylosidase by aqueous two-phase extraction", J CHROMAT B, 754(1), 2001, pp. 179-184

Abstract

A rapid process for purification of an extracellular beta -xylosidase withhigh purity was developed. The manipulation involved the precipitation of protein from culture medium and the extraction of enzyme from the resuspended elude protein solution by an aqueous-two phase separation. A linear random copolymer, PE62, with 20% ethylene oxide and 80% propylene oxide was employed in both stages of the purification. The enzyme was precipitated effectively by using 10% (w/v) PE62 and 5% (w/v) Na2HPO4. The aqueous two-phase extraction was performed with PE62 (10%)-NaH2PO4 (15%) as phase-forming reagent. SDS-PAGE analysis revealed that the purified enzyme is near homogeneity. The yield is about 100% with a purification factor of 8.8-fold. The whole process can be completed within an hour without any column chromatography. (C) 2001 Elsevier Science B.V. All rights reserved.

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Documento generato il 09/04/20 alle ore 07:28:43