Biochemical properties of lipoxygenase from opium poppy chloroplasts.

Vanko, M.; Rauová, D.; Bezáková, L.; Holková, I.; Bilka, F.; Cupáková, M.

Lipoxygenase (LOX) from opium poppy ( Papaver somniferum L.) chloroplasts was isolated and 126.1-fold purified to electrophoretic homogeneity by combination of ion-exchange chromatography on HA-Ultragel column and affinity chromatography on a linoleyl-aminopropyl agarose column. The relative molecular mass of the LOX determined by SDS-PAGE was 92 kDa. Kinetic properties of purified LOX were determined in spectrophotometric assay by using of linoleic acid (K = 1.78 mM and V = 11.4 μmol mg min) and linolenic acid (K = 1.27 mM and V = 10.2 μmol mg min). The optimum pH was 6.0 for both linoleic and linolenic acid dioxygenation catalyzed by LOX. HPLC analysis of the products revealed a dual positional specificity of linoleic acid dioxygenation at pH 6.0 with ratio of 9- and 13-hydroperoxide products being about 1:1. The activity of purified LOX was stimulated by Mg and Ca.

LIPOXYGENASES; OPIUM poppy; CHLOROPLASTS; ELECTROPHORESIS; ION exchange chromatography

Biologia Plantarum, 2012, Vol 56, Issue 1, p105

0006-3134

Academic Journal

10.1007/s10535-012-0023-4