Reversible and strong immobilization of proteins by ionic exchange on supports coated with sulfate-dextran.
New and strong ionic exchange resins have been prepared by the simple and rapid ionic adsorption of anionic polymers (sulfate-dextran) on porous supports activated with the opposite ionic group (DEAE/MANAE). Ionic exchange properties of such composites were strongly dependent on the size of the ionic polymers as well as on the conditions of the ionic coating of the solids with the ionic polymers (optimal conditions were 400 mg of sulfate-dextran 5000 kDa per gram of support). Around 80% of the proteins contained in crude extracts from Escherichia coli and Acetobacter turbidans could be adsorbed on these porous composites even at pH 7. This interaction was stronger than that using conventional carboxymethyl cellulose (CMC) and even others such as supports coated with aspartic-dextran polymer. By means of the sequential use of the new supports and supports coated with polyethyleneimine (PEI), all proteins from crude extracts could be immobilized. In fact, a large percentage (over 50%) could be immobilized on both supports. Finally, some industrially relevant enzymes (beta-galactosidases from Aspergillus oryzae, Kluyveromyces lactis, and Thermussp. strain T2, lipases from Candida antarctica A and B, Candida rugosa, Rhizomucor miehei, and Rhyzopus oryzae and bovine pancreas trypsin and chymotrypsin) have been immobilized on these supports with very high activity recoveries and immobilization rates. After enzyme inactivation, the protein could be fully desorbed from the support, and then the support could be reused for several cycles. Moreover, in some instances the enzyme stability was significantly improved, mainly in the presence of organic solvents, perhaps as a consequence of the highly hydrophilic microenvironment of the support.
Fuentes
Manuel
M
Departamento de Biocatálisis, Instituto de Catálisis-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
Pessela
Benevides C C
BC
Maquiese
Jorgette V
JV
Ortiz
Claudia
C
Segura
Rosa L
RL
Palomo
Jose M
JM
Abian
Olga
O
Torres
Rodrigo
R
Mateo
Cesar
C
Fernández-Lafuente
Roberto
R
Guisán
J M
JM
D000327:Adsorption; D016264:Dextran Sulfate / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D007475:Ion Exchange Resins; D011506:Proteins / Q000737:chemistry*
D007475:Ion Exchange Resins; D011506:Proteins; D016264:Dextran Sulfate