[Immobilization of phospholipase A2 from Central Asian cobra venom on polyamide sorbents].
The effect of the immobilization technique and the ligand nature on catalytic properties of phospholipase A2 from the cobra venom was studied. Preparations of phospholipase A2 adsorbed on and covalently bound to polyamide sorbents were obtained. The enzyme was coupled to polyamide beads modified with glutaraldehyde. In this case only 9% of the enzyme activity was retained. The enzyme adsorbed on polyamide modified with phosphatidylethanolamine retained up to 20% of the initial activity. The binding selectivity of phospholipase A2 was maximum in case of the sorbent with a binary ligand, e. g. phosphatidylethanolamine+cytotoxin, the sorbent capacity for the bound enzyme increased 2-3 times (460-600 units/g sorbent. The specific activity of the adsorbed phospholipase A2 was 17-40 units/g sorbent in contrast to 8.6 units/g sorbent for the covalently bound enzyme. Immobilization of the enzyme on polyamide sorbents resulted in changes of the pH-optimum, sensitivity to Ca2+ ions and the character of the enzyme-substrate interactions. Heart stability of the adsorbed phospholipase A2 was lower than that of the covalently bound enzyme. However, the adsorbed enzyme can be used, for example, in affinity chromatography due to its higher specific activity, selectivity and reversibility of the sorption.
Akhmedzhanov
R A
RA
Salikhova
Z T
ZT
Aripov
T F
TF
Rakhimov
M M
MM
D000327:Adsorption; D000818:Animals; D002118:Calcium / Q000378:metabolism; D004546:Elapid Venoms; D004795:Enzyme Stability; D004800:Enzymes, Immobilized; D006863:Hydrogen-Ion Concentration; D008024:Ligands; D009757:Nylons; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2; D012995:Solubility; D013696:Temperature
D004546:Elapid Venoms; D004800:Enzymes, Immobilized; D008024:Ligands; D009757:Nylons; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2; D002118:Calcium
Immobilizatsiia fosfolipazy A2 iada sredneaziatskoĭ kobry na sorbentakh poliamidnoĭ prirody.