Enzyme stabilization by glutaraldehyde crosslinking of adsorbed proteins on aminated supports.
The stabilization achieved by different immobilization protocols have been compared using three different enzymes (glutaryl acylase (GAC), D-aminoacid oxidase (DAAO), and glucose oxidase (GOX)): adsorption on aminated supports, treatment of this adsorbed enzymes with glutaraldehyde, and immobilization on glutaraldehyde pre-activated supports. In all cases, the treatment of adsorbed enzymes on amino-supports with glutaraldehyde yielded the higher stabilizations: in the case of GOX, a stabilization over 400-fold was achieved. After this treatment, the enzymes could no longer be desorbed from the supports using high ionic strength (suggesting the support-protein reaction). Modification of the enzymes immobilized on supports that did not offer the possibility of react with glutaraldehyde showed the same stability that the non modified preparations demonstrating that the mere chemical modification did not have effect on the enzyme stability. This simple strategy seems to permit very good results in terms of immobilization rate and stability, offering some advantages when compared to the immobilization on glutaraldehyde pre-activated supports.
López-Gallego
Fernando
F
Departamento de Biocatálisis, Instituto de Catálisis, CSIC, Campus Universidad Autonoma, Cantoblanco, 28049 Madrid, Spain.
Betancor
Lorena
L
Mateo
Cesar
C
Hidalgo
Aurelio
A
Alonso-Morales
Noelia
N
Dellamora-Ortiz
Gisela
G
Guisán
Jose M
JM
Fernández-Lafuente
Roberto
R
D000327:Adsorption; D001709:Biotechnology / Q000379:methods*; D003432:Cross-Linking Reagents / Q000737:chemistry; D003605:D-Amino-Acid Oxidase / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry; D005976:Glutaral / Q000737:chemistry*; D010399:Penicillin Amidase / Q000737:chemistry
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D005949:Glucose Oxidase; D003605:D-Amino-Acid Oxidase; D010399:Penicillin Amidase; C046586:glutarylamidocephalosporanic acid acylase; D005976:Glutaral