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New Cationic Exchanger Support for Reversible Immobilization of Proteins
Author(s) -
Fuentes Manuel,
Maquiese Jorgete V.,
Pessela Benevides C. C.,
Abian Olga,
FernándezLafuente Roberto,
Mateo Cesar,
Guisán J. M.
Publication year - 2008
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0342102
Subject(s) - chemistry , carboxymethyl cellulose , immobilized enzyme , aspergillus oryzae , cationic polymerization , dextran , covalent bond , adsorption , bacillus circulans , chromatography , sepharose , polymer , ionic strength , agarose , polymer chemistry , organic chemistry , enzyme , sodium , aqueous solution
New tailor‐made cationic exchange resins have been prepared by covalently binding aspartic‐dextran polymers (e.g. MW 15 000–20 000) to porous supports (aminated agarose and Sepabeads). More than 80% of the proteins contained in crude extracts from Escherichia coli and Acetobacter turbidans have been strongly adsorbed on these porous materials at pH 5. This interaction was stronger than in conventional carboxymethyl cellulose (e.g., at pH 7 and 25 °C, all proteins previously adsorbed at pH 5 were released from carboxymethyl cellulose, whereas no protein was released from the new supports under similar conditions). Ionic exchange properties of such composites were strongly dependent on the size of the aspartic‐dextran polymers as well as on the exact conditions of the covalent coating of the solids with the polymer (optimal conditions: 100 mg aspartic‐dextran 20 000/(mL of support); room temperature). Finally, some industrially relevant enzymes ( Kluyveromices lactis, Aspergillus oryzae , and Thermus sp. β‐galactosidases, Candida antarctica B lipase, and bovine pancreas trypsin and chymotrypsin) have been immobilized on these supports with very high activity recovery and immobilization rates. After enzyme inactivation, the enzyme can be fully desorbed from the support and the support could be reused for several cycles.