Fatty lipoxygenase

Lipoxygenase-Catalyzed Oxidations. Lipoxygenase-1 catalyzes the incorporation of dioxygen into polyunsaturated fatty acids possessing a l(Z),4(Z)-pentadienyi moiety to yield ( ),(Z)-conjugated hydroperoxides. A highly active preparation of the enzyme from soybean is commercially available in purified form. From a practical standpoint it is important to mention that the substrate does not need to be in solution to undergo the oxidation. Indeed, the treatment of 28 g/L of linoleic acid [60-33-3] with 2 mg of the enzyme results in (135)-hydroperoxide of linoleic acid in 80% yield... 

Applications of peroxide formation are underrepresented in chiral synthetic chemistry, most likely owing to the limited stability of such intermediates. Lipoxygenases, as prototype biocatalysts for such reactions, display rather limited substrate specificity. However, interesting functionalizations at allylic positions of unsaturated fatty acids can be realized in high regio- and stereoselectivity, when the enzymatic oxidation is coupled to a chemical or enzymatic reduction process. While early work focused on derivatives of arachidonic acid chemical modifications to the carboxylate moiety are possible, provided that a sufficiently hydrophilic functionality remained. By means of this strategy, chiral diendiols are accessible after hydroperoxide reduction (Scheme 9.12) [103,104]. 

Scheme 9.12 Soybean lipoxygenase (SBLO)-mediated oxygenation of fatty acid derivatives.

There are three groups of eicosanoids that are synthesized from C20 eicosanoic acids derived from the essential fatty acids linoleate and a-linolenate, or directly from dietary arachidonate and eicosapentaenoate (Figure 23-5). Arachidonate, usually derived from the 2 position of phospholipids in the plasma membrane by the action of phospholipase Aj (Figure 24-6)—but also from the diet—is the substrate for the synthesis of the PG2, 1X2 series (prostanoids) by the cyclooxygenase pathway, or the LT4 and LX4 series by the lipoxygenase pathway, with the two pathways competing for the arachidonate substrate (Figure 23-5). 

Lipoxygenases catalyse the regio-specific and stereoselective oxygenation of unsaturated fatty acids. The mammalian enzymes have been detected in human platelets, lung, kidney, testes and white blood cells. The leukotrienes, derived from the enzymatic action of the enzyme on arachidonic acid, have effects on neutrophil migration and aggregation, release of lysosomal enzymes, capillary permeability, induction of pain and smooth muscle contraction (Salmon, 1986). 

An example of an experiment in which LDL has been treated with 15-lipoxygenase and the oxidation monitored by the formation of conjugated diene is shown in Fig. 2.2. In the absence of transition metal, a rapid increase in absorbance occurs, with no lag phase, which ceases after a period of about 90 min under these conditions. If copper is added to promote LDL oxidation at this point, LDL treated with lipoxygenase oxidizes at a faster rate with a short lag phase when compared to the control. During this procedure there is only a minimal loss of a-tocopherol and so we may ascribe the shortened lag phase to the increase in lipid peroxides brought about by lipoxygenase treatment. A similar result was found when LDL was supplemented with preformed fatty acid hydroperoxides (O Leary eta/., 1992). 

Chamirlitrat, W. and Mason, KP. (1989). Lipid preroxyl radical intermediates in the peroxidation of polyunsaturated fatty acids by lipoxygenase. J. Biol. Chem. 264, 20968—20973. 

Fe-TPAA Fe(III)-tris[N-(2-pyridylmethyl)-2-aminoethyl] amine Fe-TPEN Fe(II)-tetrakis-N,N,N, N -(2-pyridyl methyl-2-aminoethyl)amine FFA Free fatty acids FGF Fibroblast growth factor FID Flame ionization detector FITC Fluorescein isothiocyanate FKBP FK506-binding protein FLAP 5-lipoxygenase-activating protein... 

Lipoxygenase (EC 1.13.11.12, nonheme iron dioxygenase), the substrate (polyunsaturated fatty acid) is poorly water soluble and the product (hydroperoxy-fatty acid) is hydrophilic. The reaction occurs in the aqueous phase [85,86]. 

The lipase-catalyzed fatty acid ester hydrolysis and the lipoxygenation of free polyunsaturated fatty acids are involved in the same lipid degradation pathway. They are respectively the first and second reaction in the lipoxygenase pathway (Fig. 3) [87-91]. The pathway produces volatile products of considerable importance in food technology including Cg[92, 93] or Cg- 94—96 aldehydes and alcohols from polyunsaturated fatty... 

The ability of free or immobilized lipoxygenase to introduce oxygen derived from the air into polyunsaturated fatty acids in media containing organic solvent and aqueous buffer has been investigated [9,56]. The influence of many parameters was tested upon the degree of oxygenation in biphasic systems [36,108]. 

At the end of the reaction, hydroperoxide can be easily recovered in the aqueous phase (98-99%) after its separation from the organic phase and precipitation of the enzymes. The hydroperoxides obtained are highly reactive molecules [109]. They are intermediate compounds in the lipoxygenase pathway in plants, precursors for the synthesis of hydroxy-fatty acids (i.e., ( + )-coriolic acid [38,110], and regulators of the prostaglandins biosynthesis [111-113]. 

Lipoxygenases, which catalyse the oxidation of unsaturated fatty acids containing the cis,cis-l,4-pentadiene moiety to the corresponding 1 -hydroperoxy-f rans,ds-2,4-diene (Table 2.3), are widely distributed in plants and animals. The mammalian... 

Zhuang H, Barth MM and Hildebrand DF. 1994. Packaging influenced total chlorophyll, soluble protein, fatty acid composition and lipoxygenase activity in broccoli florets. J Food Sci 59(6) 1171-1174. 

As mentioned earlier, oxidation of LDL is initiated by free radical attack at the diallylic positions of unsaturated fatty acids. For example, copper- or endothelial cell-initiated LDL oxidation resulted in a large formation of monohydroxy derivatives of linoleic and arachi-donic acids at the early stage of the reaction [175], During the reaction, the amount of these products is diminished, and monohydroxy derivatives of oleic acid appeared. Thus, monohydroxy derivatives of unsaturated acids are the major products of the oxidation of human LDL. Breuer et al. [176] measured cholesterol oxidation products (oxysterols) formed during copper- or soybean lipoxygenase-initiated LDL oxidation. They identified chlolcst-5-cnc-3(3, 4a-diol, cholest-5-ene-3(3, 4(3-diol, and cholestane-3 3, 5a, 6a-triol, which are present in human atherosclerotic plaques. 

Schnurr et al. [22] showed that rabbit 15-LOX oxidized beef heart submitochondrial particles to form phospholipid-bound hydroperoxy- and keto-polyenoic fatty acids and induced the oxidative modification of membrane proteins. It was also found that the total oxygen uptake significantly exceeded the formation of oxygenated polyenoic acids supposedly due to the formation of hydroxyl radicals by the reaction of ubiquinone with lipid 15-LOX-derived hydroperoxides. However, it is impossible to agree with this proposal because it is known for a long time [23] that quinones cannot catalyze the formation of hydroxyl radicals by the Fenton reaction. Oxidation of intracellular unsaturated acids (for example, linoleic and arachidonic acids) by lipoxygenases can be suppressed by fatty acid binding proteins [24]. 

Long ago it was noticed that the baking quality of white flour improved with storage for 1-2 months. This effect occurred more rapidly if the flour was exposed to the air. During storage, initially the level of free fatty acids increases, presumably owing to lipolytic activity. Lipoxygenase activity then produces oxidised fatty acids as the proportion of linoleic and linolenic acids falls while the number of -S-S- bonds decreases. 

Flavor is one of the major characteristics that restricts the use of legume flours and proteins in foods. Processing of soybeans, peas and other legumes often results in a wide variety of volatile compounds that contribute flavor notes, such as grassy, beany and rancid flavors. Many of the objectionable flavors come from oxidative deterioration of the unsaturated lipids. The lipoxygenase-catalyzed conversion of unsaturated fatty acids to hydroperoxides, followed by their degradation to volatile and non-volatile compounds, has been identified as one of the important sources of flavor and aroma components of fruits and vegetables. An enzyme-active system, such as raw pea flour, may have most of the necessary enzymes to produce short chain carbonyl compounds. 

Lipoxygenase (linoleate oxygen oxidoreductase) catalyzes the hydroperoxidation of fatty acids containing a methylene-interrupted conjugated diene system. The degradation of the hydroperoxides results in the formation of numerous secondary products (46-48). 

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