Lipoxygenase Activity in Pig Muscle:  Purification and Partial Characterization

Abstract

Lipoxygenase from Iberian pig Biceps femoris muscle was purified in a process that involves two successive chromatographic steps on DEAE-Sephadex and phenyl-Sepharose CL4B. The purified enzyme had a final specific activity of 52 mU/mg, a purification factor of 3250, a molecular weight of 90 kDA, and a maximum activity at pH 5.5. The K M values obtained for linoleic acid (K M = 0.28 mM), arachidonic acid (K M = 3.8 mM), and linolenic acid (K M = 0.43 mM) reveal a preferential use of linoleic acid as substrate. When purified enzyme was incubated in the presence of linoleic acid, two main products were identified by direct-phase HPLC: 9-hydroperoxy octadecadienoic acid and 13-hydroperoxy octadecadienoic acid in the ratio of 45:55. The presence of lipoxygense activity suggest a possible participation of this enzyme in the biogenesis of flavor and aroma in hams from Iberian pigs. Keywords: Lipoxygenase; purification; hams; Iberian pig; lipid oxidation

Chinese red pepper (Zanthoxylum bungeanum Maxim.) leaf extract as natural antioxidants in salted silver carp (Hypophthalmichthys molitrix) in dorsal and ventral muscles during processing

“…The linoleic acid substrate solution was prepared following the method of Gata, Pinto, and Macías (1996) by combining 140 mg of linoleic acid (Sigma) with 5 mL of deionized water containing 180 mL of Tween 20. The solution was brought to pH 9.0 by adding 2 N NaOH.…”

Examination of the effects of temperature and pressure on lipoxygenase activities in pork using response surface methodology

“…A linoleic acid solution was prepared following the method of Gata et al (2). Linoleic acid (140 mg) was dissolved in 5 mL of deionized water containing 180 µL of Tween 20.…”

“…LOXs are widely distributed in plants, animals, and microorganisms; however, current research has focused mainly on plants, particularly legume plants, because the content of LOXs in plants is higher than in animals and microorganisms. LOXs in animal-derived foods are rarely studied and most previous studies only focused on purification and enzymatic properties (1)(2)(3)(4)(5)(6)(7)(8). LOXs play an important role in flavor formation of meat products, it can catalytically oxidize polyunsaturated fatty acids to generate numerous different hydroperoxides, in conjunction with many different decomposition pathways, leading to a large number of volatile compounds (9)(10)(11).…”

Catalytic mechanisms of metmyoglobin on the oxidation of lipids in phospholipid liposome model system

“…Lipoxygenase-like (LOX-like) activity of metMb (1 mg/mL) was measured by the method of Gata, Pinto, and Macias (1996) with some modifications. Linoleic acid (10 mM) in 0.02 M NaOH solution emulsified with Tween-20 was used as a substrate solution, which was flushed with and kept under nitrogen.…”