Chemical modification of protein surfaces to improve their reversible enzyme immobilization on ionic exchangers.
The enzyme penicillin G acylase (PGA) is not adsorbed at pH 7 on DEAE- or PEI-coated supports, neither is it adsorbed on carboxymethyl (CM)- or dextran sulfate (DS)-coated supports. The surface of the enzyme was chemically modified under controlled conditions: chemical amination of the protein surface of carboxylic groups (using soluble carbodiimide and ethylendiamine) and chemical succinylation (using succinic anhydride) of amino groups. The full chemical modification produced some negative effects on enzyme stability and activity, although partial modification (mainly succinylation) presented negligible effects on both enzyme features. The chemical amination of the protein surface permitted the immobilization of the enzyme on CM- and DS-coated support, while the chemical succinylation permitted the enzyme immobilization on DEAE- and PEI-coated supports. Immobilization was very strong on these supports, mainly in the polymeric ones, and dependent on the degree of modification, although the enzymes still can be desorbed after inactivation by incubation under drastic conditions. Moreover, the immobilization on ionic polymeric beds allowed a significant increase in enzyme stability against the inactivation and inhibitory effects of organic solvents, very likely by the promotion of a certain partition of the organic solvent out of the enzyme environment. These results suggest that the enrichment of the surface of proteins with ionic groups may be a good strategy to take advantage of the immobilization of industrial enzymes via ionic exchange on ionic polymeric beds.
Montes
Tamara
T
Departamento de Biocatalisis, Instituto de Catalisis, CSIC, Campus Universidad Autonoma, Cantoblanco, 28049 Madrid, Spain.
Grazu
Valeria
V
López-Gallego
Fernando
F
Hermoso
Juan A
JA
Guisan
José M
JM
Fernandez-Lafuente
Roberto
R
D000327:Adsorption; D004800:Enzymes, Immobilized / Q000737:chemistry*; D007475:Ion Exchange Resins; D008958:Models, Molecular; D010399:Penicillin Amidase / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*
D004800:Enzymes, Immobilized; D007475:Ion Exchange Resins; D011506:Proteins; D010399:Penicillin Amidase