Immobilization of catalase via adsorption onto metal-chelated affinity cryogels
Yükleniyor...
Tarih
2012
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Elsevier
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
A poly (acrylamide-allylglycidyl ether) [p(AAm-AGE)] cryogel was prepared by radical polymerization of acrylamide (AAm) and allylglycidyl ether (AGE). Cibacron Blue F3GA (CB) was covalently attached to the p(AAm-AGE) cryogel via the reaction between the chloride groups of the reactive dyes and the epoxide groups of the AGE. The CB-attached p(AAm-AGE) cryogel was chelated with Fe3+ ions. This immobilized metal ion affinity chromatography (IMAC) cryogel carrying 25.8 +/- 2.0 mu mol Fe3+ ions was used in adsorption studies to interrogate the effects of pH, protein initial concentration, flow rate, temperature and ionic strength on enzyme activity. Maximum adsorption capacities were found to be 75.7 +/- 1.2 mg/g for p(AAm-AGE)-CB-Fe3+ cryogels and 60.6 +/- 1.0 mg/g for p(AAm-AGE)-CB cryogels, respectively. The adsorbed amounts of catalase per unit mass of cryogel reached a plateau value at about 1.5 mg/mL at pH 6.0. The K-m values were found to be 0.73 +/- 0.02 g/L for the free catalase and 0.18 +/- 0.02 g/L for the immobilized catalase. The V-max value of free catalase (2.0 x 10(3) U/mg enzyme) was found to be lower than that of the immobilized catalase (2.5 x 10(3) U/mg enzyme). It was also observed that the enzyme could be repeatedly adsorbed and desorbed onto the p(AAm-AGE)-CB-Fe3+ cryogel. (C) 2011 Elsevier Ltd. All rights reserved.
Açıklama
Anahtar Kelimeler
Catalase, Cryogel, IMAC, Enzyme Immobilization, Adsorption, Dye Ligand
Kaynak
Process Biochemistry
WoS Q Değeri
N/A
Scopus Q Değeri
Q1
Cilt
47
Sayı
1
