Polyketone polymer: A new support for direct enzyme immobilization

Abstract

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 • 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.

Enzyme immobilization: an update

“…An innovative immobilization process has been recently proposed [46] which utilizes polyketone polymer, a completely new support. Polyketone polymer -[-CO-CH 2 CH 2 -] n -, obtained by copolymerization of ethane and carbon monoxide, has been utilized for immobilization of three different enzymes, one peroxidase from horseradish and two amine oxidases from bovine serum and lentil seedlings.…”

“…Polyketone polymers are obtained by palladiumcatalyzed copolymerization of ethane and carbon monoxide [46]. Due to the close presence of polar keto groups, the polymers present a high liquid volume value (8.2 ml/g for a polymer not exceeding 25,000 Da), larger than twice than that shown by commercial sepharose, because the high possibility of hydrogen bonds between the keto groups and the water molecules and no restriction due to cross linking are present.…”

“…Due to the close presence of polar keto groups, the polymers present a high liquid volume value (8.2 ml/g for a polymer not exceeding 25,000 Da), larger than twice than that shown by commercial sepharose, because the high possibility of hydrogen bonds between the keto groups and the water molecules and no restriction due to cross linking are present. Water accessible carbonyl groups were quantified in about 10 meq/g polymer, and the apparent concentration of the immobilized enzyme on the polyketone was more than 2.36 mg/ml [46]. Biopolymers A variety of biopolymers, mainly water-insoluble polysaccharides such as cellulose, starch, agarose, carragenans, and chitosan, have been widely used as supports for immobilizing enzymes (Fig.…”

“…In addition, the mild operating conditions of enzymatic processes mean that they can be operated in relatively simple and totally controlled equipment. In short, they reduce environmental drawbacks of manufacturing by Applicable for large enzymes, peroxidase, and amine oxidases [46] Classical covalent immobilization…”

Studies on the activity and stability of immobilized horseradish peroxidase on poly(ethylene terephthalate) grafted acrylamide fiber

“…Support material, which plays an important role in the utility of an immobilized enzyme, should be readily available and non-toxic and also should provide a large surface area suitable for enzyme reaction, and substrate and product transport with least-diffusional restriction [8]. Some organic and inorganic supports like glass beads [9], polyketone [10], chitosan [11] and aluminum oxide/polyethyleneimine composite [12] have been used for the immobilization of peroxidases.…”