Kinetic properties of lipoxygenase from desert truffle (Terfezia claveryi Chatin) ascocarps: effect of inhibitors and activators

There is very little information available on the kinetic characteristics of fungal lipoxygenases (LOXs) because most data on the mechanism of this enzyme concern soybean LOX. In this paper, the kinetic properties of LOX from Terfezia claveryi Chatin ascocarps were studied for the first time. The enzyme did not show the "substrate aggregation-dependent activity" described for other LOXs and presented a K(m) for linoleic acid of 41 microM at pH 7.0. The effect of different inhibitors was also studied. The enzyme presented the characteristic lag phase of other LOXs, and the influence of different factors on its duration was analyzed. The lag period was reduced not only by the product of the reaction (13-HPOD) but also by 9-HPOD. Calculation of the activation constant is proposed for the first time as a useful tool for the characterization of LOX because this method makes it possible to quantify the effectiveness of different hydroperoxides as LOX activators. The activation constants obtained were 0.3 and 6.4 microM for 13- and 9-HPOD, respectively; thus, the product of the reaction was approximately 21-fold more effective than 9-HPOD as a T. claveryi LOX activator.     

Dual positional and stereospecificity of lipoxygenase isoenzymes from germinating barley (green malt): biotransformation of free and esterified linoleic acid

The lipoxygenase isoenzymes LOX1 and LOX2 from green malt were separated by isoelectric focusing, and their catalytic properties regarding complex lipids as substrates were characterized. The regio- and stereoisomers of hydroperoxy octadecadienoates (HPODE) resulting from LOX1 and LOX2 enzymatic transformations of linoleic acid, methyl linoleate, linoleic acid glycerol esters monolinolein, dilinolein, and trilinolein, and 1-palmitoyl-2-linoleoyl-glycero-3-phosphocholine (PamLinGroPCho) were determined. In addition, biotransformations of polar and nonpolar lipids extracted from malt were performed with LOX1 and LOX2. The results show that LOX2 catalyzes the oxidation of esterified fatty acids at a higher rate and is more regioselective than LOX1. The dual position specificity of LOX2 (9-HPODE:13-HPODE) with trilinolein as the substrate (6:94) was higher than the resultant ratio (13:87) when free linoleic acid was transformed. A high (S)-enantiomeric excess of 13-HPODE was analyzed with all esterified substrates confirming the formation of 13-HPODE through the LOX2 enzyme; however, 9-HPODE detected after LOX2 biotransformations showed (R)-enantiomeric excesses. PamLinGroPCho was oxygenated by LOX1 with the highest regio- and stereoselectivities among the applied substrates.     

A simple and efficient system for green note compound biogenesis by use of certain lipoxygenase and hydroperoxide lyase sources

Six-carbon (C(6)) aldehydes and alcohols are important components of the aroma and flavor of fruits and vegetables. Soybean lipoxygenase (LOX) isozyme LOX 3 was reported not only to produce less 13-hydroperoxides, precursors of C(6) aldehydes, but also to convert them to ketodiene products. Here, we examined the effects of LOX 3 on hexenal formation from linolenic acid homogenized with watermelon 13-hydroperoxide lyase (HL)-overexpressing Nicotiana tabacum leaves and soybean acetone powder. Compared to the wild type, which contains LOXs 1, 2, and 3, the elimination of LOX 3 in LOX 1 + 2 facilitates greater production of hexenals. The use of LOX 2 alone yielded the highest hexenal production, while a two-step conversion was required for LOX 1 to produce hexenals at high levels due to different pH optima of the enzymes involved. These results clearly demonstrate that the soybeans lacking LOX 3 in combination with watermelon HL-overexpressing leaf tissues greatly enhance hexenal formation.     

Eggplant lipoxygenase (Solanum melongena): product characterization and effect of physicochemical properties of linoleic acid on the enzymatic activity

Lipoxygenase (LOX) from eggplant (Solanum melongena L. cv. Belleza negra) was partially purified, and the products and kinetics of the enzyme were studied. Linoleic acid (LA) was the best substrate for this enzyme. Product analysis by HPLC and GC/MS revealed that, at its pH optimum (pH 7.0), the enzyme converted LA almost totally into the 9-hydroperoxy isomer, whereas the 13-hydroperoxy isomer was only a minor product. At this pH, the enzyme had K(m) and V(max) values for LA of 1.4 microM and 2.2 micromol min(-1) (mg of protein)(-1), respectively, when the monomeric form of LA was used as substrate. The dependence of eggplant LOX activity on the physicochemical properties of LA was also studied. Experiments revealed that LA aggregates were used more efficiently than monomeric LA as substrate. The apparent substrate cooperativity observed may be due to the different activities exhibited toward monomers and aggregates. This result can be interpreted as a substrate-aggregation dependent activity.     

桃脂氧合酶(LOX)基因家族cDNA全长克隆与序列分析

根据脂氧合酶保守序列设计的简并引物从桃（瑞蟠5号）果肉总RNA中扩增出795bp的脂氧合酶（lipoxygenase,LOX）基因的保守序列,并结合GeneBank中公开的LOX基因的EST序列经Blast比对分析、序列组装之后设计基因特异引物（GSP）。利用RACE技术共获得3条不同的桃LOX基因cDNA全长序列,分别命名为PpLox1-3。序列分析结果表明,这3个桃LOX基因家族成员与其他植物的LOX基因尽管在核酸水平上的差异比较大,但在蛋白水平上却有较高的同源性。通过对桃LOX基因蛋白序列和其他植物LOX进行系统进化树分析,发现PpLox1属于13-LOX类型,PpLox2-3属于9-LOX类型。     

Lipoxygenase distribution in coffee (Coffea arabica L.) berries

In this paper lipoxygenase (LOX) presence was investigated in coffee berries to determine its involvement in lipid degradative metabolism of plants grown in organic and conventional cultivations. An immunochemical analysis has evidenced a ca. 80 kDa protein, cross-reacting with an anti-LOX antibody, only in the pulp fraction of berries obtained from plants of both cultivations. LOX activity in this fraction could be monitored either as conjugated diene formation or reaction products (determined by HPLC) and was mainly associated with a heavy membrane fraction (HMF, enriched in tonoplast, endoplasmic reticulum, plasma membrane, and mitochondria) and a light membrane fraction (LMF, enriched in plasma membrane and endoplasmic reticulum, with low levels of tonoplast and mitochondria). The LOX activity of LMF from berries of both cultivations showed an optimum at pH 8.0. The HMF exhibited a different activity peak in samples from conventional (pH 8.0) and organic (pH 5.5) cultures, suggesting the presence of different isoenzymes. These findings were also confirmed by variation of the ratio of 9- and 13-hydroperoxides in organic (1:1) and conventional cultivations (1:10), indicating that the organic one was subjected to an oxidative stress in the coffee pulp fraction leading to the expression of an acidic LOX. Such de novo synthesized LOX activity could be responsible for the production of secondary metabolites, which may interfere with the organoleptic profile of coffee.     

Lipoxygenase from banana leaf: purification and characterization of an enzyme that catalyzes linoleic acid oxygenation at the 9-position

The objective of the present study was to purify and characterize the lipoxygenase (LOX) from banana leaf (Giant Cavendishii, AAA), an unutilized bioresource. LOX was extracted, isolated, and purified 327-fold using 25-50% saturation of ammonium sulfate fractionation, hydroxyapatite column separation, and gel filtration on Superdex 200. The molecular mass of the purified LOX was 85 kDa, K(m) was 0.15 mM, and V(max) was 2.4 microM/min.mg using linoleic acid as substrate. Triton X-100 was required in the extraction medium; otherwise, no LOX activity was detected. LOX activity increased with the concentration of Triton X-100 with an optimum at 0.1%. The optimal pH of the purified LOX from banana leaf was 6.2, and optimal temperature was 40 degrees C. The LOX showed the highest reactivity toward 18:2 followed by 18:3 and 20:4. A very low reaction rate was observed toward 20:5 and 22:6. On the basis of retention time in normal phase HPLC, the products of 18:2 or 18:3 catalyzed by purified LOX were hydroperoxyoctadecadienoic acid or hydroperoxyoctadecatrienoic acid. It seems that 9-LOX is the predominant enzyme in banana leaf. Banada leaf dried at 110 degrees C for 2 h developed algal aroma. Banana leaf extract stored at 10 degrees C for 12 h formed an oolong tea-like flavor. Banana leaf extract reacted with 18:2 or soybean oil pretreated with bacterial lipase produced green and melon-like aroma, whereas the same reaction with 18:3 produced a sweet, fruity, cucumber-like flavor note.     

Thermal inactivation kinetics of recombinant proteins of the lipoxygenase pathway related to the synthesis of virgin olive oil volatile compounds

The aim of this work was to characterize the thermal inactivation parameters of recombinant proteins related to the biosynthesis of virgin olive oil (VOO) volatile compounds through the lipoxygenase (LOX) pathway. Three purified LOX isoforms (Oep2LOX1, Oep1LOX2, and Oep2LOX2) and a hydroperoxide lyase (HPL) protein (OepHPL) were studied. According to their thermal inactivation parameters, recombinant Oep1LOX2 and Oep2LOX2 could be identified as the two LOX isoforms active in olive fruit crude preparations responsible for the synthesis of 13-hydroperoxides, the main substrates for the synthesis of VOO volatile compounds. Recombinant Oep2LOX1 displayed a low thermal stability, which suggests a weak actuation during the oil extraction process considering the current thermal conditions of this industrial process. In addition, recombinant OepHPL could be identified as the HPL activity in crude preparations. The thermal stability was the highest among the recombinant proteins studied, which suggests that HPL activity is not a limiting factor for the synthesis of VOO volatile compounds.     

Comparative study of hops-containing products on human cytochrome P450-mediated metabolism

The potential for 15 different ales (6), ciders (2 apple and 1 pear), and porters (6) and 2 non-alcoholic products to affect cytochrome P450 (CYP)-mediated biotransformation and P-glycoprotein-mediated efflux of rhodamine was examined. As in our previous study, a wide range of recovered nonvolatile suspended solids dry weights were noted. Aliquots were also found to have varying effects on biotransformation and efflux. Distinct differences in product ability to affect the safety and efficacy of therapeutic products confirmed our initial findings that some porters (stouts) have a potential to affect the safety and efficacy of health products metabolized by CYP2D6 and CYP3A4 isozymes. Most products, except 2 of the ciders and the 2 non-alcoholic products, also have the potential to affect the safety of CYP2C9 metabolized medications and supplements. Further studies are required to determine the clinical significance of these findings.     

Biosynthesis of trans-2-hexenal in response to wounding in strawberry fruit

Wounded strawberry fruit produces a diverse group of volatile compounds including aldehydes, alcohols, and esters derived from the lipoxygenase (LOX) and hydroperoxide lyase (HPL) pathways. Because the wound volatiles may play an important role in plant-fungal interaction, the goal of this study was to develop a greater understanding about the biosynthesis of the major wound volatile, trans-2-hexenal (t-2-H), produced by strawberry fruit upon wounding. To that end, composition and quantity of total and free fatty acids of control and wounded strawberry fruit were analyzed. In addition, activities of the key enzymes, LOX and HPL, and production of C6 aldehydes were determined. Intact strawberry fruit did not produce detectable t-2-H which is derived from alpha-linolenic acid (18:3). However, in response to wounding by bruising, strawberry fruit emitted t-2-H and its precursor cis-3-hexenal (c-3-H). The level of total lipid 18:3 in the fruit increased 2-fold in response to wounding, whereas free 18:3 declined slightly ( approximately 30%). At 10 min following wounding, fruit exhibited a 25% increase in LOX activity, which leads to the production of 13-hydroperoxyoctadecatrienoic acid (13-HPOT) from 18:3. The activity of HPL, which catalyzes formation of cis-3-hexenal from 13-HPOT, increased 2-fold by 10 min after wounding. Thus, during a 15 min period after wounding, free 18:3 substrate availability and the activity of two key enzymes, LOX and HPL, changed in a manner consistent with increased c-3-H and t-2-H biosynthesis.     

Formation of volatile compounds in model experiments with crude leek (Allium ampeloprasum Var. Lancelot) enzyme extract and linoleic acid or linolenic acid

Three continuous assays are described for lipoxygenase (LOX), hydroperoxide lyase (HPL) and alcohol dehydrogenase (ADH) in leek tissue. The catalytic activity of LOX showed significant difference (significance level 5%) between linolenic acid (9.43 x 10(-)(4) katals per kg protein) and linoleic acid (2.53 x 10(-)(4) katals per kg protein), and the pH-optimum of LOX was 4.5-5.5 against linoleic acid. The catalytic activity of HPL was statistically the same for 9-(S)-hydroperoxy-(10E,12Z)-octadecadienoic acid (1.01 x 10(-)(2) katals per kg protein) and 13-(S)-hydroperoxy-(9Z,11E)-octadecadienoic acid (7.69 x 10(-)(3) katals per kg protein). ADH showed a catalytic activity of 5.01 x 10(-)(4) katals/kg of protein toward hexanal. Model experiments with crude enzyme extract from leek mixed with linoleic acid or linolenic acid demonstrated differences in the amount of produced aroma compounds. Linoleic acid resulted in significantly most hexanal, heptanal, (E)-2-heptenal, (E)-2-octenal, (E,E)-2,4-decadienal, pentanol, and hexanol, whereas linolenic acid resulted in significantly most (E)-2-pentenal, (E)-2-hexenal, (E,Z)-2,4-heptadienal, (E,E)-2,4-heptadienal, and butanol. Leek LOX produced only the 13-hydroperoxide of linoleic acid and linolenic acid.     

Oxygenation of arachidonoyl lysophospholipids by lipoxygenases from soybean, porcine leukocyte, or rabbit reticulocyte

Oxygenation of arachidonoyl lysophosphatidylcholine (lysoPC) or arachidonoyl lysophosphatidic acid (lysoPA) by lipoxygenase (LOX) was examined. The oxidized products were identified by HPLC/UV spectrophotometry/mass spectrometry analyses. Straight-phase and chiral-phase HPLC analyses indicated that soybean LOX-1 and rabbit reticulocyte LOX oxygenated arachidonoyl lysophospholipids mainly at C-15 with the S form as major enantiomer, whereas porcine leukocyte LOX oxygenated at C-12 with the S form. Next, the sequential exposure of arachidonoyl-lysoPC to soybean LOX-1 and porcine leukocyte LOX afforded two major isomers of dihydroxy derivatives with conjugated triene structure, suggesting that 15(S)-hydroperoxyeicosatetraenoyl derivatives were converted to 8,15(S)-dihydroxyeicosatetraenoyl derivatives. Separately, arachidonoyl-lysoPA, but not arachidonoyl-lysoPC, was found to be susceptible to double oxygenation by soybean LOX-1 to generate a dihydroperoxyeicosatetraenoyl derivative. Overall, arachidonoyl lysophospholipids were more efficient than arachidonic acid as LOX substrate. Moreover, the catalytic efficiency of arachidonoyl-lysoPC as substrate of three lipoxygenases was much greater than that of arachidonoyl-lysoPA or arachidonic acid. Taken together, it is proposed that arachidonoyl-lysoPC or arachidonoyl-lysoPA is efficiently oxygenated by plant or animal lipoxygenases, C12- or C15-specific, to generate oxidized products with conjugated diene or triene structure.     

Lipoxygenase and hydroperoxide lyase activities in ripening strawberry fruits

The enzymes lipoxygenase and hydroperoxide lyase have been identified in strawberry (Fragariax ananassa Duch.) var. Camarosa. Their subcellular localization, substrate preference, and product specificity were determined in mature strawberry fruits. The activity of both enzymes was located mainly in the microsomal fraction. Linolenic acid was the preferred substrate for strawberry lipoxygenase, forming 13- and 9-hydroperoxides of this acid in the proportion 70:30. The strawberry hydroperoxide lyase cleaves 13-hydroperoxide of linoleic (13% relative activity) and linolenic (100% relative activity) acids to form hexanal and (3Z)-hexenal, respectively. Both enzyme activities and endogenous content of volatile aldehydes formed by sequential action of lipoxygenase-hydroperoxide lyase were evaluated during strawberry development and ripening. A sequential enzymatic pathway for the formation of green odor compounds in strawberry is proposed.     

Co-oxidation of beta-carotene catalyzed by soybean and recombinant pea lipoxygenases

A number of products including apocarotenal, epoxycarotenal, apocarotenone, and epoxycarotenone generated by lipoxygenase (LOX) catalyzed co-oxidation of beta-carotene have been tentatively identified through the use of GC/MS and HPLC combined with photodiode array detection. Because of the large number of high molecular weight products detected and their probable chemical structures, a co-oxidation mechanism is proposed that involves random attack along the alkene chain of the carotenoid by a LOX-generated linoleoylperoxyl radical. It is suggested that a direct release from the enzyme of the radical, which initiates the co-oxidation of beta-carotene, is greater for pea LOX-3 than for pea LOX-2 or soybean LOX-1. It is proposed that further products may be formed by free radical propagated reactions and that the formation of 1,10- and 1,14-dicarbonyl compounds may arise by secondary oxidation of the primary products.     

Acid-catalyzed hydrolysis of alcohols and their beta-D-glucopyranosides

The hydrolysis, in model wine at pH 3, of the allylic, homoallylic, and propargylic glycosides, geranyl-beta-D-glucopyranoside, [3'-(1' '-cyclohexenyl)-1'-methyl-2'-propynyl]-beta-D-glucopyranoside, (3'RS, 9'SR)-(3'-hydroxy-5'-megastigmen-7-yn-9-yl)-beta-D-glucopyra noside, (3',5',5'-trimethyl-3'-cyclohexenyl)-beta-D-glucopyranoside, E-(7'-oxo-5',8'-megastigmadien-3'-yl)-beta-D-glucopyranoside (3-hydroxy-beta-damascone-beta-D-glucopyranoside), and their corresponding aglycons has been studied. In general, aglycons were more rapidly converted to transformation products than were the corresponding glucosides. Glycoconjugation of geraniol in grapes is a process that reduces the flavor impact of this compound in wine, not only because geraniol is an important flavor component of some wines but also because the rate of formation of other flavor compounds from geraniol during bottle-aging is reduced. However, when flavor compounds such as beta-damascenone are formed in competition with flavorless byproducts, such as 3-hydroxy-beta-damascone, by acid-catalyzed hydrolytic reactions of polyols, then glycoconjugation is a process that could enhance as well as suppress the formation of flavor, depending on the position of glycosylation. (3'RS, 9'SR)-(3'-Hydroxy-5'-megastigmen-7'-yn-9'-yl)-beta-D-glucopy ranoside hydrolyzed more slowly but gave a higher proportion of beta-damascenone in the products than did the aglycon at 50 degrees C. Reaction temperature also effected the relative proportion of the hydrolysis products. Accelerated studies do not parallel natural processes precisely but only approximate them.     

Metabolism and lack of DNA reactivity of the mycotoxin ochratoxin a in cultured rat and human primary hepatocytes

It is still unclear whether the carcinogenic mycotoxin ochratoxin A (OTA) is bioactivated to DNA-binding metabolites in rodents and humans. Therefore, we have incubated cultured rat and human primary hepatocytes with noncytotoxic concentrations of (3)H-OTA ranging from 10(-7) to 10(-5) M for 8 h and determined its metabolism and covalent DNA binding. In rat hepatocytes, OTA was metabolized to small amounts of three products, which were further studied by electrospray ionization (ESI)-MS/MS techniques. In addition to 4-hydroxy-OTA, which is a known product of OTA biotransformation, two novel metabolites were detected and tentatively identified as hexose and pentose conjugates of OTA. The in vitro induction with 3-methylcholanthrene (3MC) increased the formation of 4-hydroxy-OTA but did not alter the formation of the conjugated metabolites. No covalent binding of (3)H-OTA or its metabolites to DNA was observed in rat hepatocytes with or without 3MC induction with a limit of detection of 2 adducts per 10(9) nucleotides. However, the cellular ratio of reduced glutathione to oxidized glutathione was significantly decreased by treatment with OTA. In cultured human hepatocytes, (3)H-OTA was only very poorly metabolized, and no covalent DNA binding was observed. In conclusion, the results of this in vitro study do not support the notion that OTA has the potential to undergo metabolic activation and form covalent DNA adducts in rodents and humans.     

Reactivity of epicatechin in aqueous glycine and glucose maillard reaction models: quenching of C2, C3, and C4 sugar fragments

Mechanisms of how epicatechin alters the pathways of the Maillard reaction were investigated. Carbon-13 and nitrogen-15 labeling studies were utilized to define the reactivity of epicatechin with glucose, glycine, and/or reaction products in an aqueous model (pH 7, 125 degrees C for 30 min) via GC, GC/MS and HPLC/MS analysis. Quantification of the volatile reaction product isotopomers by GC/MS from a 1:1 labeled to unlabeled glucose (carbohydrate module labeling technique) plus glycine model system indicated the formation of 2,3-butanedione and acetol were primarily formed via intact C4 and C3 sugar fragments, whereas pyrazine, methylpyrazine, 2,5-dimethylpyrazine, 2,3,5-trimethylpyrazine, and cyclotene were primarily formed via intact C2/C2, C2/C3, C3/C3, C3/C3, and C3/C3 sugar fragment pairs, respectively. The formation of these seven compounds was also reported by GC analysis to be dramatically inhibited when epicatechin was added to the glucose/glycine model system (observed 9-113-fold reduction). HPLC/MS analysis of both the glucose-labeled and glycine-labeled model systems with and without epicatechin indicated that epicatechin reacted directly with C2, C3, and C4 sugar fragments, while epicatechin did not report any direct reactivity with glycine. In conclusion, the quenching of sugar fragmentation products via epicatechin was correlated with the observed inhibition on volatile compound formation when epicatechin was added to a glucose/glycine aqueous reaction model system.     

Comparison between discontinuous and continuous lactose conversion processes for the production of prebiotic galacto-oligosaccharides using beta-galactosidase from Lactobacillus reuteri

Galacto-oligosaccharide (GOS) formation from lactose in discontinuous and continuous modes of conversion was investigated using beta-galactosidase (beta-gal) from Lactobacillus reuteri. A continuous stirred tank reactor (CSTR) with an external crossflow membrane was set up, and continuous GOS production was analyzed and compared to the batchwise formed GOS product. Marked differences were detected for the two reactor setups. Above 65% lactose conversion, the GOS yield was lower for the CSTR due to a lower content of tri- and tetrasaccharides in the reaction mixture. In the CSTR, beta-gal from L. reuteri showed up to 2-fold higher specificity toward the formation of beta-(1-->6)-linked GOS, with beta-D-Galp-(1-->6)-D-Glc and beta-D-Galp-(1-->6)-D-Gal being the main GOS components formed under these conditions. This could be used to synthesize more defined GOS products.     

Degradation of alachlor in natural and sludge-amended soils, studied by gas and liquid chromatography coupled to mass spectrometry (GC-MS and HPLC-MS)

Alachlor [2-chloro-N-(2,6-diethylphenyl)-N-(methoxymethyl)acetamide] is an herbicide used worldwide. The relative rates of disappearance of alachlor, the formation kinetics of alachlor ethane sulfonic acid (ESA), and the formation of other degradation products in two different soils (a soil with natural organic matter and a sludge-amended soil) has been studied. For such a purpose, soil samples were spiked with alachlor at 2.5 mg kg(-1), concentration generally applied in agricultural soils, and were submitted to sunlight, simulating natural field conditions. Extracts were analyzed by GC-MS and HPLC-MS in scan mode. A good correlation was observed between both techniques, and HPLC-MS allowed the determination of two eluting peaks corresponding to the two stereoisomeric forms of alachlor ESA. Degradation of alachlor in the two soils followed first-order kinetics. Half-life in the natural soil was 4.2 +/- 0.1 days, and half-life in the sludge-amended soil was 5.8 +/- 0.8 days. The higher half-life observed in the sludge-amended soil was attributed to the higher sorption of alachlor to this soil compared to the natural soil. The degradation of alachlor in both soils gave rise to the production of alachlor ESA. Its concentration increased during the incubation period, and after 27 days, its concentration was about 0.59 mg kg(-1) in the natural soil and 0.37 mg kg(-1) in the sludge-amended soil. The other two alachlor transformation products were identified using GC-MS, and the abundance of these degradation products increased while alachlor was degraded.     

Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds.

Lipoxygenase (LOX) is thought to play an important role in the formation of desirable or undesirable flavor and aroma in many plant products. In rice seeds, LOX activity is localized in the bran fraction and LOX-3 is the major isozyme component. We used gas chromatography-mass spectrometry to determine whether the degree of staleness in the flavor of stored brown rice was related to the presence of LOX-3. We found that the amount of hexanal, pentanal, and pentanol in normal raw LOX-3 rice markedly increased during storage at 35 degrees C. That in LOX-3-less rice increased slightly but was a third to a fifth that of normal LOX-3 rice. In cooked rice, the amount of these components from glutinous rice exceeded that in nonglutinous rice, and that in normal LOX-3 rice exceeded that in LOX-3-less rice. These results indicate that the stale flavor production in LOX-3-less rice during storage is less than that in normal LOX-3 rice.