Effect of titanium on the activity of lipoxygenase

Lipoxygenase was extracted from untreated tomato fruits and those treated with titanium ascorbate (TITAVIT) at the last stage of ripening. It was found that titanium (Ti) had a activating effect on lipoxygenase so that the development of red pigments was inhibited to some extent. The addition of Titavit to the reaction mixture resulted in a lag period of about 1 minute to lipoxygenase‐catalyzed linoleic acid oxidation, whereas, incubation of this complex with enzyme extract for 10 minutes significantly increased enzyme activity. Activation of lipoxygenase by Ti was confirmed when addition of titanium chloride also increased the enzyme activity. Titanium also increased the stability of lipoxygenase preparations when kept under refrigeration.     

Inactivation of soybean trypsin inhibitors and lipoxygenase by high-pressure processing

Trypsin inhibitors (TIA), one of the antinutritional factors of soy milk, are usually inactivated by heat treatment. In the current study, high-pressure processing (HPP) was evaluated as an alternative for the inactivation of TIA in soy milk. Moreover, the effect of HPP on lipoxygenase (LOX) in whole soybeans and soy milk was studied. For complete LOX inactivation either very high pressures (800 MPa) or a combined temperature/pressure treatment (60 degrees C/600 MPa) was needed. Pressure inactivation of TIA was possible only in combination with elevated temperatures. For TIA inactivation, three process parameters, temperature, time, and pressure, were optimized using experimental design and response surface methodology. A 90% TIA inactivation with treatment times of <2 min can be reached at temperatures between 77 and 90 degrees C and pressures between 750 and 525 MPa.     

Effect of high-pressure treatment on the carotenoid composition and the radical scavenging activity of persimmon fruit purees

The carotenoid composition of persimmon fruit purees of two cultivars, cvs. Rojo Brillante and Sharon, grown in Spain was determined by HPLC to assess the effects of high-pressure processing on some sensory (carotenoids), nutritional (provitamin A value), and health-related (radical-scavenging capacity) parameters. Total carotenoid content was higher in untreated Rojo Brillante puree (22. 11 microg g(-)(1)) than in untreated Sharon puree (15.22 microg g(-)(1)). Purees of both untreated cultivars showed similar carotenoid patterns after saponification with beta-cryptoxanthin, beta-carotene, and zeaxanthin as the main pigments. A high content of lycopene was quantified in Rojo Brillante (5.34 microg g(-)(1)), whereas only traces were detected in Sharon. The provitamin A value, reported as retinol equivalents (RE), was in untreated Rojo Brillante puree (77 RE/100 g) similar to that of Sharon (75 RE/100 g). Scavenging free radical capacity, measured as antiradical efficiency (AE), showed in untreated Rojo Brillante puree a value (12.14 x 10(-)(3)) 8.5 times higher than that in untreated Sharon (1. 42 x 10(-)(3)). Nonuniform behavior of high-pressure treatment was detected. Pressure treatments at 50 and 300 MPa/15 min/25 degrees C for Rojo Brillante and at 50 and 400 MPa/15 min/25 degrees C for Sharon increased the amount of extractable carotenoids (9-27%), which are related with the increase of vitamin A value (75-87 RE/100 g). No correlation with the increase of AE (from 1.42 x 10(-)(3) to 16.73 x 10(-)(3) and 19.58 x 10(-)(3)) after some pressure treatments (150 and 300 MPa/15 min/25 degrees C) was found.     

Effect of high-pressure treatment on the texture of cherry tomato

The effect of high-pressure treatment (200-600 MPa for 20 min) on the texture of cherry tomatoes and on the key softening enzymes (pectinmethylesterase and polygalacturonase) was investigated. When subjected to high-pressure treatment whole cherry tomatoes showed increasing textural damage with increasing pressures up to 400 MPa. However, treatment at pressures above 400 MPa (500-600 MPa) led to less apparent damage than treatment at 300 and 400 MPa; the tomatoes appearing more like the untreated samples. These visual changes were reflected in the texture (firmness) and amount of cell rupture in the tomatoes, with the least firmness and the most cell rupture being seen after treatment at 400 MPa. Light and scanning electron microscopy supported these observations. Although a sample of purified commercial pectinmethylesterase was partially inactivated at pressures above 200 MPa, irrespective of pH (4-9), in the whole cherry tomatoes no significant inactivation was seen even after treatment at 600 MPa, presumably because other components in the tomato offered protection or the isoenzymes were different. Polygalacturonase was more susceptible to pressure, being almost totally inactivated after treatment at 500 MPa. It is concluded that the textural changes in tomato induced by pressure involve at least two related phenomena. Initially, damage is caused by the greater compressibilty of the gaseous phase (air) compared to liquid-solid components, giving rise to a compact structure which, on pressure release, is damaged as the air rapidly expands, leading to increases in membrane permeability. This permits egress of water, and the damage also enables enzymatic action to increase, causing further cell damage and softening. The major enzyme involved in the further softening is polygalacturonase, which is inactivated at 500 MPa and above, and not pectinmethylesterase, which in the whole fruit, is barotolerant.     

Effect of pepper lipoxygenase activity and its linked reactions on pigments of the pepper fruit.

The products formed during the enzymatic reaction catalyzed by the lipoxygenase of pepper (variety Agridulce) have in vitro a strong destructive action on the carotenoid pigments of the fruit. When conditions and proportions of enzyme and pigments are similar to those found in the fruit, and at a reaction temperature of 20 degrees C, almost 30% of the pigments are destroyed after 24 h of reaction. Of this amount, 2.5% is due to autoxidation of pigments, 4. 5% to oxidation induced by the presence of linoleic under saturating conditions, and the remaining 22% to the presence in the medium of reaction products of the lipoxygenase-catalyzed reaction. When the enzyme acts under substrate-saturating conditions, the rate of pigment destruction by lipoxygenase can be considered maximal at the experimental temperature. The fact that in vitro pepper lipoxygenase induces a heavy destruction of pigments and that, in vivo, its activity remains almost constant during over-ripening could explain why up to 40% of the pigment content in some varieties is lost during the postharvest period.     

Colorimetric method for the determination of lipoxygenase activity

A colorimetic assay for lipoxygenase activity has been developed. The assay is based on the detection of the lipoxygenase reaction product, linoleic acid hydroperoxide, by the oxidative coupling of 3-methyl-2-benzothiazolinone (MBTH) with 3-(dimethylamino)benzoic acid (DMAB) in a hemoglobin-catalyzed reaction. This test reaction is rapid and sensitive, and it offers advantages over other methods for detecting lipoxygenase activity. The assay is capable of detecting activity in a number of crude vegetable homogenates and should be particularly useful where a rapid visual determination of lipoxygenase activity is desired.     

Oxidation of dietary polyphenolics by hydroperoxidase activity of lipoxygenase

Lipoxygenase, in the presence of hydrogen peroxide, produces the oxidative decomposition of quercetin, naringenin, and resveratrol, known antioxidant molecules. Quercetin was the molecule more efficiently oxidized, followed by resveratrol and naringenin. When this molecule was incubated in the presence of GSH, a quinoid derivative was produced. This compound was not obtained in the presence of naringenin or resveratrol, suggesting that in the presence of hydrogen peroxide and lipoxygenase, quercetin may be oxidized to a prooxidant species. When hydrogen peroxide was substituted by hydroperoxy linoleic acid, the same oxidative process was observed. This means that in food products in which lipoxygenase and linoleic acid are presents, quercetin may be oxidized to prooxidant species; in contrast, naringenin and resveratrol may constitute a valid additive for the prevention of the oxidative degradation of foods.     

Effect of high-pressure processing on activity and structure of alkaline phosphatase and lactate dehydrogenase in buffer and milk

Changes in the activity and structure of alkaline phosphatase (ALP) and L-lactate dehydrogenase (LDH) were investigated after high pressure processing (HPP). HPP treatments (206-620 MPa for 6 and 12 min) were applied to ALP and LDH prepared in buffer, fat-free milk, and 2% fat milk. Enzyme activities were measured using enzymatic assays, and changes in structure were investigated using far-ultraviolet circular dichroism (CD) spectroscopy and dynamic light scattetering (DLS). Kinetic data indicated that the activity of ALP was not affected after 6 min of pressure treatments (206-620 MPa), regardless of the medium in which the enzyme was prepared. Increasing the processing time to 12 min did significantly reduce the activity of ALP at 620 MPa (P < 0.001). However, even the lowest HPP treatment of 206 MPa induced a reduction in LDH activity, and the course of reduction increased with HPP treatment until complete inactivation at 482, 515, and 620 MPa. CD data demonstrated a partial change in the secondary structure of ALP at 620 MPa, whereas the structure of LDH showed gradual denaturation after exposure at 206 MPa for 6 min, leading to a random coil structure at both 515 and 620 MPa. DLS results indicated aggregation of ALP only at HPP treatment of 206 MPa and not above and enzyme precipitation as well as aggregation at 345, 415, 482, and 515 MPa. The loss of LDH activity with increasing pressure and time treatment was due to the combined effects of denaturation and aggregation.     

Regulation of lipoxygenase activity by polyunsaturated lysophosphatidylcholines or their oxygenation derivatives

Lysophosphatidylcholines (lysoPCs) have been known to play a role as lipid mediators in various cellular responses. In this study, we examined whether lysoPC containing linoleoyl, arachidonoyl, or docosahexaenoyl groups or their peroxy derivatives affect lipoxygenase (LOX)-catalyzed oxygenation of native substrates. First, arachidonoyl lysoPC and docosahexaenoyl lysoPC were found to inhibit potato 5-LOX-catalyzed oxygenation of linoleic acid (LA) in a noncompetitive type with Ki values of 0.38 and 1.90 microM, respectively. Likewise, arachidonoyl lysoPC and docosahexaenoyl lysoPC also inhibited 5-LOX activity from rat basophilic leukemia cells-2H3 (RBL-2H3) with IC50 values (50% inhibitory concentration) of 18.5 +/- 3.06 and 30.6 +/- 1.06 microM, respectively. Additionally, arachidonoyl lysoPC and docosahexaenoyl lysoPC also inhibited 15-LOX activity from RBL-2H3 with IC50 values of 16.6 +/- 1.3 and 24.1 +/- 2.4 microM, respectively. In a separate experiment, where lysoPC peroxides were tested for the effect on soybean LOX-1, 15(S)-hydroperoxy-5,8,11,13-eicosatetraenoyl lysoPC and 17(S)-hydroperoxy-4,7,10,13,15,19-docosahexaenoyl lysoPC potently inhibited soybean LOX-1 activity with Ki values of 6.8 and of 1.54 microM, respectively. In contrast, 13(S)-hydroperoxy-9,11-octadecadienoyl lysoPC was observed to stimulate soybean LOX-1-catalyzed oxygenation of LA markedly with AC50 value (50% activatory concentration) of 1.5 microM. Taken together, it is proposed that lysoPCs containing polyunsaturated acyl groups or their peroxy derivatives may participate in the regulation of LOX activity in biological systems.     

Resveratrol is a potent inhibitor of the dioxygenase activity of lipoxygenase

Resveratrol is a naturally occurring phytoalexin, present in grapes and other food products, with important antioxidant properties. Although still under debate, it is generally assumed that resveratrol has protective effects against heart diseases and probably tumor development. Lipoxygenase is a dioxygenase with peroxidase activity involved in the synthesis of mediators in inflammatory, atherosclerotic, and carcinogenic processes. Lipoxygenase activity is also involved in the generation of flavors and aromas in foods from animal or vegetal sources. The results presented here show that resveratrol was a potent inhibitor of the dioxygenase activity of lipoxygenase, with an IC(50) = 13 microM. Simultaneously, resveratrol was oxidized by the peroxidase activity of lipoxygenase with a V(max) = 0.28 microM min(-1) and a k(M) = 16.6 microM. Furthermore, oxidized resveratrol was as efficient a lipoxygenase inhibitor as in its reduced form. From the data obtained it can be concluded that both resveratrol and its oxidized form can act as inhibitors of the dioxygenase activity of lipoxygenase. In contrast, the hydroperoxidase activity of lipoxygenase was not inhibited by resveratrol. These results suggest that resveratrol may be used as an antioxidant food additive and as a pharmacological agent to prevent the generation of eicosanoids involved in pathological processes.     

Effect of titanium ascorbate on the lipoxygenase pathway of tomato and red pepper seedlings

The effect of titanium applied as titanium ascorbate (Titavit^®) on the lipoxygenase (LOX) pathway of tomato and red pepper seedlings was studied. Seeds were germinated in solutions containing titanium at 5 and 10 ppm. The uptake of titanium was significant and occurred at a similar amount for both tomato and red pepper. LOX and hydroperoxide decomposing (HPD) activities were determined in the crude extracts. The results showed that LOX activity in the extract of Titavit‐germinated seedlings was significantly higher than that of the control, whereas no effect was observed on the activity of HPD enzymes. It is suggested that activation of LOX by Titavit was due to a chemical interaction of titanium within the reaction mixture than to an induction of the biosyntheis of the enzyme.     

Effect of soybean lipoxygenase on volatile generation and inhibition of Aspergillus flavus mycelial growth

Volatiles generated from lipoxygenase (LOX) normal and LOX deficient soybean (Glycine max) varieties with and without added lipase inhibited Aspergillus flavus mycelial growth and aflatoxin production. Soybean volatiles were analyzed using a solid phase microextraction (SPME) method combined with gas chromatography-mass spectrometry (GC-MS). Twenty-one compounds, including 11 aldehydes, three alcohols, four ketones, one furan, one alkane, and one alkene were detected in the LOX normal soybean line. However, only nine volatile compounds were observed in the LOX deficient soybean variety. The antifungal aldehydes hexanal and (E)-2-hexenal were observed in both LOX normal and LOX deficient lines and were detected at significantly higher amounts in soybean homogenate with added lipase. These aldehydes may be formed through alternate pathways, other than the LOX pathway, and may account for the inhibition of A. flavus growth observed. Other volatiles detected, particularly the ketones and alcohols, may contribute to the antifungal activity observed in both LOX normal and LOX deficient soybean lines. These results suggest that other factors, other than LOX activity, may better explain why soybeans are generally not as severely affected by A. flavus and aflatoxin contamination as other oilseed crops.     

热辅助超高压对复合甜面酱品质特性的影响

该文分别从流变、色差、风味及脂肪酸组成等方面展开试验,旨在研究热辅助超高压技术对甜面酱感官品质等品质特性的影响。结果表明,复合甜面酱经热辅助超高压处理后,黏度降低,并随着压力增加,黏度逐渐下降。经300 MPa和400 MPa处理后的样品的黏度变化小于500 MPa处理的样品。经过加压处理后,甜面酱的色差值(ΔE)发生显著变化,棕色指数(browning index,BI)下降,但亮度(L*)和红绿色度值(a*)无显著变化(P0.05)。复合甜面酱中的挥发性风味物质以醛类、酯类为主,酯类含量在高压处理后减少。此外,400 MPa的热辅助超高压处理并没有对脂肪酸的组成和含量造成较大的影响。由此得知,400 MPa、45℃、10 min的热辅助超高压处理可以较好地保护复合甜面酱质地、色泽、风味等方面的品质。为热辅助超高压技术对复合调味酱的进一步研究及工业化生产提供了理论依据。     

Structural changes of microbial transglutaminase during thermal and high-pressure treatment

The activity of microbial transglutaminase (MTG) and the corresponding secondary structure, measured by circular dichroism (CD), was analyzed before and after treatment at different temperatures (40 and 80 degrees C) and pressures (0.1, 200, 400, 600 MPa). Irreversible enzyme inactivation was achieved after 2 min at 80 degrees C and 0.1 MPa. Enzyme inactivation at 0.1, 200, 400, and 600 MPa and 40 degrees C followed first-order kinetics. The enzyme showed residual activity of 50% after 12 min at 600 MPa and 40 degrees C. Mobility of aromatic side chains of the enzyme molecule was observed in all temperature- and/or pressure-treated samples; however, high-pressure treatment at 600 MPa induced a loss of tertiary structure and a significant decrease in the alpha-helix content. The relative content of beta-strand substructures was significantly increased after 30 min at 600 MPa and 40 degrees C or 2 min at 0.1 MPa and 80 degrees C. We conclude that the active center of MTG, which is located in an expanded beta-strand domain, is resistant to high hydrostatic pressure and pressure-induced inactivation is caused by destruction of alpha-helix elements with a corresponding influence on the enzyme stability in solution.     

Effect of heat treatment on the antioxidant activity of extracts from citrus peels

The effect of heat treatment on the antioxidant activity of extracts from Citrus unshiu peels was evaluated. Citrus peels (CP) (5 g) were placed in Pyrex Petri dishes (8.0 cm diameter) and heat-treated at 50, 100, or 150 degrees C for 10, 20, 30, 40, 50, and 60 min in an electric muffle furnace. After heat treatment, 70% ethanol extract (EE) and water extract (WE) (0.1 g/10 mL) of CP were prepared, and total phenol contents (TPC), radical scavenging activity (RSA), and reducing power of the extracts were determined. The antioxidant activities of CP extracts increased as heating temperature increased. For example, heat treatment of CP at 150 degrees C for 60 min increased the TPC, RSA, and reducing power of EE from 71.8 to 171.0 microM, from 29.64 to 64.25%, and from 0.45 to 0.82, respectively, compared to non-heat-treated control. In the case of WE from CP heat-treated at the same conditions (150 degrees C for 60 min), the TPC, RSA, and reducing power also increased from 84.4 to 204.9 microM, from 15.81 to 58.26%, and from 0.27 to 0.96, respectively. Several low molecular weight phenolic compounds such as 2,3-diacetyl-1-phenylnaphthalene, ferulic acid, p-hydroxybenzaldoxime, 5-hydroxyvaleric acid, 2,3-diacetyl-1-phenylnaphthalene, and vanillic acid were newly formed in the CP heated at 150 degrees C for 30 min. These results indicated that the antioxidant activity of CP extracts was significantly affected by heating temperature and duration of treatment on CP and that the heating process can be used as a tool for increasing the antioxidant activity of CP.     

Functional properties, lipoxygenase activity, and health aspects of Lupinus albus protein isolates

To utilize lupin seeds for food and pharmaceutical applications, lupin seeds were pretreated to remove oil using hexane or carbon dioxide. Two types of lupin protein isolate were prepared. Both types of protein isolate showed good foaming activity, comparable to egg white. Protein isolate extracted under acid conditions showed higher foaming activity than protein isolate extracted at neutral pH. The lipoxygenase activity was much reduced in both of the protein isolates. The protein isolate extracted at neutral pH showed a stronger angiotensin converting enzyme inhibition than the protein isolate extracted under acidic pH. In contrast, the protein isolate extracted under acid conditions had a greater sodium cholate binding capacity, comparable to that of cholestyramine. Lupin samples showed less DPPH radical scavenging activity than deoiled soybean. The deoiling method did not affect the functional properties, lipoxygenase activity, angiotensin converting enzyme inhibition, sodium cholate binding, and radical scavenging activity.     

Determination of lipoxygenase activity in plant extracts using a modified ferrous oxidation-xylenol orange assay

The ferrous oxidation-xylenol orange (FOX) assay method for determination of lipid hydroperoxides is based on that under acidic conditions Fe2+ is oxidized to Fe3+, which then oxidizes xylenol orange to a product that absorb at 550 nm. The procedure has been adapted for determination of lipoxygenase activity in plant extracts. This enzyme is responsible for generation of off-flavors in vegetal foods, bleaching of pigments, and a lot of oxidative degradations. It is of interest to check the initial lipoxygenase activity in vegetal foods before the processing, using an assay that is rapid, reproducible, and easily adaptable to high throughput format. The enzymatic assay is based on a discontinuous determination of lipoxygenase activity using the FOX reagent for colorimetric determination of hydroperoxides accumulated in the medium by a period of incubation that is established by the addition of the extract (start of the reaction) and the addition of FOX reagent (finish of the reaction). The procedure is capable of detecting lipoxygenase activity in a number of vegetable homogenates, being especially useful for a rapid visual evaluation of this enzymatic activity.     

超高压处理对砀山梨汁中过氧化物酶活性的影响

该文研究了超高压处理对砀山梨汁中过氧化物酶活性的影响。试验压力为0.1～500 MPa，温度为20～60℃。此外，考察了不同pH值(3～7)和保压时间(2～34 min)超高压处理对酶活性的影响。试验结果表明：在处理温度为50℃、保压时间为10 min和梨汁pH 5的条件下，300 MPa以下压力范围内高压处理酶被激活，其活性增加；大于300 MPa时酶的活性随压力增大而下降。高压处理时，温度低于40℃对酶的活性影响不大；有效影响高压处理的最小温度为40℃。保压时间超过10 min后时间延长对酶的活性影响     

Effect of mild-heat and high-pressure processing on banana pectin methylesterase: a kinetic study

Pectin methylesterase (PME) was extracted from bananas and purified by affinity chromatography. The thermal-high-pressure inactivation (at moderate temperature, 30-76 degrees C, in combination with high pressure, 0.1-900 MPa) of PME was investigated in a model system at pH 7.0. Under these conditions, the stable fraction was not inactivated and isobaric-isothermal inactivation followed a fractional-conversion model. At lower pressure (< or =300-400 MPa) and higher temperature (> or =64 degrees C), an antagonistic effect of pressure and heat was observed. Third-degree polynomial models (derived from the thermodynamic model) were successfully used to describe the heat-pressure dependence of the inactivation rate constants.