title: Immobilization of β-galactosidase on modified polypropilene membranes.
abstract: A new immobilized system: β-galactosidase-modified polypropylene membrane was created. It was obtained 13 different carriers by chemical modification of polypropylene membranes by two stages. The first stage is treatment with K(2)Cr(2)O(7) to receive carboxylic groups on membrane surface. The second stage is treatment with different modified agents ethylendiamine, hexamethylenediamine, hydrazine dihydrochloride, hydroxylamine, o-phenylenediamine, p-phenylenediamine, N,N'-dibenzyl ethylenediamine diacetate to receive amino groups. The quantity of the amino groups, carboxylic groups and the degree of hydrophilicity of unmodified and modified polypropilene membranes were determined. β-Galactosidase was chemically immobilized on the obtained carries by glutaraldehyde. The highest relative activity of immobilized enzyme was recorded at membrane modified with 10% hexamethylenediamine (Membrane 5) - 92.77%. The properties of immobilized β-galactosidase on different modified membranes - pH optimum, temperature optimum, pH stability and thermal stability were investigated and compared with those of free enzyme. The storage stability of all immobilized systems was studied. It was found that the most stable system is immobilized enzyme on Membrane 5. The system has kept 90% of its initial activity at 300th day (pH=6.8; 4°C). The stability of the free and immobilized β-galactosidase on the modified membrane 5 with 10% HMDA in aqueous solutions of alcohols - mono-, diol and triol was studied. The kinetics of enzymatic reaction of free and immobilized β-galactosidase on the modified membrane 5 at 20°C and 40°C and at the optimal pH for both forms of the enzyme were investigated. It was concluded that the modified agent - hexamethylenediamine, with long aliphatic chain ensures the best immobilized β-galactosidase system.
authors.affiliation: Department of Biotechnology and Food Products, University of Rousse Angel Kanchev, Subsidiary of the University of Ruse in Razgrad, Aprilsko vastanie Blvd. 47, Razgrad 7200, Bulgaria.
authors.forename: Nastya
authors.initials: N
authors.lastname: Vasileva
authors.forename: Vladislav
authors.initials: V
authors.lastname: Iotov
authors.forename: Yavor
authors.initials: Y
authors.lastname: Ivanov
authors.forename: Tzonka
authors.initials: T
authors.lastname: Godjevargova
authors.forename: Nana
authors.initials: N
authors.lastname: Kotia
mesh_terms: D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000378:metabolism; D008567:Membranes, Artificial; D008958:Models, Molecular; D011126:Polypropylenes / Q000737:chemistry*; D011487:Protein Conformation; D001616:beta-Galactosidase / Q000737:chemistry* / Q000378:metabolism
chemical_list: D004800:Enzymes, Immobilized; D008567:Membranes, Artificial; D011126:Polypropylenes; D001616:beta-Galactosidase