Polyketone polymer: a new support for direct enzyme immobilization.
Polyketone polymer -[-CO-CH(2)-CH(2)-](n)-, obtained by copolymerization of ethene and carbon monoxide, is utilized for immobilization of three different enzymes, one peroxidase from horseradish (HRP) and two amine oxidases, from bovine serum (BSAO) and lentil seedlings (LSAO). The easy immobilization procedure is carried out in diluted buffer, at pH 7.0 and 3 degrees C, gently mixing the proteins with the polymer. No bifunctional reagents and spacer arms are required for the immobilization, which occurs exclusively via a large number of hydrogen bonds between the carbonyl groups of the polymer and the -NH groups of the polypeptidic chain. Experiments demonstrate a high linking capacity of polymer for BSAO and an extraordinary strong linkage for LSAO. Moreover, activity measurements demonstrate that immobilized LSAO totally retains the catalytic characteristics of the free enzyme, where only a limited increase of K(M) value is observed. Finally, the HRP-activated polymer is successfully used as active packed bed of an enzymatic reactor for continuous flow conversion and flow injection analysis of hydrogen peroxide containing solutions.
Agostinelli
E
E
Department of Biochemical Sciences A. Rossi Fanelli, University of Rome La Sapienza and CNR, Biology and Molecular Pathology Institutes, Rome, Italy.
Belli
F
F
Tempera
G
G
Mura
A
A
Floris
G
G
Toniolo
L
L
Vavasori
A
A
Fabris
S
S
Momo
F
F
Stevanato
R
R
D001709:Biotechnology; D002248:Carbon Monoxide / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005030:Ethylenes / Q000737:chemistry; D006860:Hydrogen Bonding; D006863:Hydrogen-Ion Concentration; D007659:Ketones / Q000737:chemistry*; D015394:Molecular Structure; D009195:Peroxidase / Q000737:chemistry; D011108:Polymers / Q000737:chemistry*
D004800:Enzymes, Immobilized; D005030:Ethylenes; D007659:Ketones; D011108:Polymers; D002248:Carbon Monoxide; C036216:ethylene; D009195:Peroxidase