Identification of lipoxygenase expressing ω-6 dioxygenase activity by means of liquid chromatography

Authors: Peter Hoffman;  Mária Cupáková 1;  Drahomíra Rauová 1;  Renáta Kollárová;  Mária Pekárová;  Marek Obložinský;  Lýdia Bezáková
Authors‘ workplace: Katedra bunkovej a molekulárnej biológie liečiv, Farmaceutická fakulta Univerzita Komenského ;  Toxikologické a antidopingové centrum, Farmaceutická fakulta, Univerzita Komenského v Bratislave, Slovenská republika 1
Published in: Čes. slov. Farm., 2012; 61, 53-59
Category: Original Articles


Lipoxygenases (LOX) represent a family of lipid peroxidising enzymes which catalyse the reaction of achiral polyunsaturated fatty acids by oxygen forming chiral peroxide products possessing high positional stereospecific purity. The four double bonds of arachidonic acid, the main substrate of animal LOX, present the position for a wide range of enzymatic modifications enabling eicosanoid creation, unique molecules with biological significance. In this study, lipoxygenase from rat lung cytoplasma was isolated and purified to 40-fold by combining hydrophobic and gel chromatography. The forming positional specific fatty acid hydroxyl-isomers were separated on a non-polar system (RP-HPLC) and identified on a polar adsorbent (SP-HPLC). In the purified enzyme, dual positional specificity was demonstrated by the production of 12- and 15-HETE in the ratio of 1,0 : 1,38, which responds to the product spectrum of mammalian 15-LOX-1.

lipoxygenase • purification, HPLC analysis • 15-LOX-1


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