Thermal Inactivation Kinetics of Peroxidase and Lipoxygenase from Broccoli, Green Asparagus and Carrots

ABSTRACT: Thewermal inactivation curves for peroxidase (POD) and lipoxygenase (LOX) in broccoli (florets), green asparagus (tip and stem), and carrots (cortex and core) extracts were determined in the range of 70 to 95 °C for 0 to 600 s. The capillary tube method was used to obtain quasi-isothermal conditions. The kinetics of both enzymes showed a biphasic first-order model, while at 70 °C, LOX in asparagus showed a monophasic first-order behavior. LOX activity was not detected for carrots. Kinetic parameters, k and Ea , were determined for heat-labile and heatresistant isoenzyme fractions. Additionally, initial and residual activities for both enzymes within tissue sections showed a different distribution and heat stability.     

Thermal Inactivation Kinetics of Wheat Germ Lipoxygenase

Thermal inactivation curves for wheat germ lipoxygenase (LPO) in partially purified and crude extracts were determined in capillary tubes at 60–68°C. The biphasic curves fitted a two-fraction first order model suggesting the presence of 2 groups of isozymes. At 60°C, the inactivation rate constants were 9.112 × 10^?-⁵ set^?1 and 9.174 × 10^?-⁶ set^?1 respectively, for thermolabile phase I and thermostable phase II in the partially purified extract, a difference of one order of magnitude. For the temperature change from 60 to 68°C, the rate constants increased by three orders of magnitude, implying a very high sensitivity (for LPO inactivation in partially purified extract ΔH^?= 646261 J.mole^?1, ΔS^?= 1619 J.mole^?1.K^?1 for phase I, ΔH^?= 546099 J.mole-^l, ΔS^?= 1298 J.mole^?1.K-1^′ for phase II) to heat by both phases, although phase I was clearly the least stable.     

鲜枣脂氧合酶酶学特性及热失活动力学研究

以亚油酸为底物,采用紫外分光光度法测定鲜枣脂氧合酶(lipoxygenase,LOX)的酶学特性及热失活动力学性质。考察了温度、p H、金属离子及络合物对LOX活性的影响,并建立了LOX酶促反应动力学及热失活动力学参数。结果表明:鲜枣LOX的最大吸收波长为265 nm;最适温度和p H分别为40℃、7.0;在p H值为5.0~6.0的环境中,LOX活性保持相对稳定;酶促反应动力学符合米氏方程所描述的单底物酶促反应动力学,其Km和Vmax分别为0.14 mmol/L、0.26 U/min,鲜枣LOX与底物亚油酸的亲和力较好;且LOX的热失活遵循一级反应动力学规律,反应活化能为105.20 k J/mol;2 mmol/L Cu2+和10 mmol/L Cu2+、Ca2+、Mn2+对LOX酶活力有激活作用(P0.05),而2 mmol/L Mn2+、Zn2+、Mg2+和10 mmol/L Mg2+,EDTA对LOX活性具有抑制作用(P0.05)。该试验为鲜枣加工贮藏中LOX活性控制提供了有益的数据参考。     

几种物质对龙眼果肉中过氧化物酶和脂肪氧合酶的影响

采用单因素及均匀设计多因素实验 ,研究究了蔗糖、CaCl2 、柠檬酸、抗坏血酸、没食子酸对龙眼果肉中过氧化物酶 (POD)、脂肪氧合酶 (LOX)的影响。结果表明 :上述试剂达到一定浓度时均对POD、LOX产生抑制作用 ,其中柠檬酸、抗坏血酸和没食子酸之间存在协同增效作用 ,当 3者的质量浓度分别为 6 .1、0 5、0 0 5g/L时 ,龙眼果肉中POD、LOX相对酶活分别为 16 7%、0 0 5 % ,抑制率分别为 83 3%、99 5 %     

Peroxidase and Lipoxygenase Influence on Stability of Polyunsaturated Fatty Acids in Sweet Corn (Zea mays L.) during Frozen Storage

The effect of blanching treatments and packaging materials on lipoxygenase (LOX) and peroxidase (POD) activity and fatty acid stability of two cultivars of sweet corn (Jubilee and GH-2684) were evaluated during 9 mo storage at ?20°C. Complete inactivation of LOX and POD was obtained with 9 and 15 min of steam blanching, respectively. Relative fatty acid content revealed no change in fatty acid composition during storage. Control of degradation of polyunsaturated fatty acids (PUFA) did not depend on oxygen permeability of different packaging materials. Blanching had little effect on PUFA degradation after 9 mo storage.     

莴笋烫漂过程中过氧化物酶失活动力学模型的建立

为有效预测一定时间、温度条件下的过氧化物酶活力,避免过度烫漂导致莴笋品质的损坏,本研究对75～95℃条件下莴笋烫漂过程中过氧化物酶失活的动力学特性建立数学模型。利用一级动力学模型和Weibull分布模型进行拟合,以决定系数(R2)和卡方(χ2)为评价指标,分析了两种模型的拟合效果。此外以VC含量为指标优化烫漂工艺条件,将模型应用于实际预测中。结果表明：Weibull分布模型的拟合效果较好,R2均大于0.9900,χ2均小于0.0024,高温短时条件下的VC含量损失较少。     

Pressure-Temperature Inactivation of Lipoxygenase in Green Peas ((Pisum sativum): A Kinetic Study

ABSTRACT Kinetics of lipoxygenase (LOX) inactivation in green peas (Pisum sativum) by temperature and/or pressure was studied at different levels of food complexity, namely in green pea juice and in intact vegetables (in situ study). Pea LOX in both matrices was irreversibly inactivated either by thermal or by combined pressure-temperature treatment, and the inactivation followed a first- order reaction. Pressure levels below 200 MPa retarded thermal inactivation of LOX at temperatures above 60 °C. Moreover, above 60 °C, enzyme activation under pressure occurred prior to inactivation. At elevated pressure, LOX inactivation rate could be accelerated by a temperature increase above 10 °C as well as by a temperature decrease below 10 °C. The pressure-temperature dependence of the LOX inactivation rate constants both in green pea juice and in intact green peas was described successfully using the same mathematical model. On the basis of this mathematical model, an iso-rate contour diagram for LOX inactivation in green pea juice and in intact green peas was constructed.     

猪肉脂肪氧化酶在高压结合热处理过程中失活动力学模型

为了解猪肉高压加工过程中脂肪氧化酶(lipoxygenase,LOX)活性的变化规律,以猪背最长肌为原料,经不同压力(200~700 MPa)、温度(20~60℃)结合处理后,研究LOX失活常数的变化规律,并分别用Arrhenius和Eyring方程为基础来建立LOX失活的动力学模型。结果表明,当处理温度不变时,失活常数随压力的增加而增加;同样,当处理压力不变时,失活常数随温度升高而增加。Arrhenius和Eyring方程分别适用于整个研究的温度区间和压力区间,分别以两者为基础可以建立LOX失活的动力学方程,两个方程的拟合程度均较高,决定系数分别达到0.989和0.965。因此在一定的处理温度(20~60℃)和压力(200~700 MPa)范围内,猪肉高压结合热处理过程中LOX活性的变化可以用以Arrhenius和Eyring方程为基础建立的数学模型进行预测。     

橄榄多酚氧化酶和过氧化物酶的抑制剂筛选及其热失活动力学

本项目研究了七种抑制剂(分别为亚硫酸钠、明矾、O-异抗坏血酸、EDTA-2Na、Ca Cl2、多聚磷酸钠、柠檬酸亚锡二钠)对橄榄多酚氧化酶和过氧化物酶活力的影响,并采用两段模型分析了高温处理对橄榄果实多酚氧化酶(PPO)和过氧化物酶(POD)的钝化动力学。结果表明,橄榄中酚类化合物氧化分解与POD和PPO活性有关,在特定的环境条件下,O-异抗坏血酸和柠檬酸亚锡二钠抑制橄榄PPO酶和POD酶活性效果最好,譬如,其浓度为0.20 g/L时,可完全抑制橄榄PPO酶和POD酶活力;热烫时间和温度对橄榄果实PPO和POD活性影响较大,并随着热烫时间和热烫温度的增加,橄榄果实PPO和POD活性呈现先下降后处于平稳的趋势,故高温处理对橄榄PPO和POD的钝化过程符合两段模型。     

Kinetics of Peroxidase Inactivation in Carrot Juice Treated with Pulsed Electric Fields

Peroxidase activity accounts for quality losses in many plant‐based foods. The paper provides insight into the inactivation kinetics of peroxidase (POD) in carrot juice treated with pulsed electric fields (PEF). Juice samples were subjected to electric field intensities of 20 to 35 kV/cm for 300 to 2000 μs. Up to 93% of the initial activity was inactivated after treating at 35 kV/cm for 1500 μs. POD activity inactivation correlated well with the increase in energy density input. A first‐order fractional conversion model best fitted the experimental results. Other kinetic approaches such as the Fermi's model can be used to estimate residual POD activity values in treated juices as a function of electric field strength. Practical Application : Pulsed electric fields (PEF) are a processing technology that can be used for the pasteurization of liquid food products. Peroxidase activity inhibition is required in carrot juices to prevent undesirable quality losses, such as discoloration, flavor changes, and loss of nutrients. The most significant processing parameters ruling POD inactivation in PEF‐treated carrot juice are identified and mathematical modeling of experimental data is conducted.     

Thermal and High-Pressure Inactivation of Tomato Polygalacturonase: A Kinetic Study

ABSTRACT: Polygalacturonase (PG) was extracted from tomato fruit. The inactivation kinetics of the PG crude extract was investigated under isothermal and isobaric-isothermal conditions. Thermal inactivation of tomato PG in the temperature range 58 to 65 °C could be described by a fractional conversion model, and in the temperature range from 70 to 90 °C by a biphasic model. Pressure-temperature inactivation (range 300 to 600 MPa/5 to 50 °C) of tomato PG could be described by a fractional conversion model, which points to first-order inactivation kinetics of a pressure-sensitive enzyme fraction and to the occurrence of a pressure-stable PG fraction. A mathematical model describing the influence of pressure and temperature on the inactivation rate constant was formulated.     

pH Affects the Thermal Inactivation Parameters of R-Phycoerythrin from Porphyra yezoensis

ABSTRACT: R-phycoerythrin (PE), a protein that fluoresces in the visible range, was purified from Porphyra yezoensis. PE had a molecular mass of 292 kDa and an isoelectric point of 4.1 to 4.2. Thermal inactivation parameters of PE, calculated on the basis of fluorescence loss, were determined under different pH conditions. PE was more thermostable between pH 5.0 and 8.0, and became more heat sensitive at pH 4.0 and 10.0. PE at pH 6.0 had the highest D value (12258.7 min) at 70 °C. The z values of PE increased from 4.58 °C at pH 5.0 to 9.15 °C at pH 9.0. PE could be used as a time-temperature integrator by adjusting the inactivation kinetics of PE to match those of target microorganisms in a thermal process.     

Kinetics of Thermal Inactivation of Peroxidase and Color Degradation of African Cowpea (Vigna unguiculata) Leaves

Cowpea leaves form an important part of the diet for many Kenyans, and they are normally consumed after a lengthy cooking process leading to the inactivation of peroxidase (POD) that could be used as an indicator for the potential shelf life of the vegetables. However, color degradation can simultaneously occur, leading to poor consumer acceptance of the product. The kinetics of POD in situ thermal (for thermal treatments in the range of 75 to 100 °C/120 min) inactivation showed a biphasic first‐order model, with Arrhenius temperature dependence of the rate constant. The kinetic parameters using a reference temperature (T_ref) of 80 °C were determined for both the heat‐labile phase (k_ref = 11.52 ± 0.95 × 10^?2 min^?1 and E_a of 109.67 ± 6.20 kJ/mol) and the heat‐stable isoenzyme fraction (k_ref = 0.29 ± 0.07 × 10^?2 min^?1 and E_a of 256.93 ± 15.27 kJ/mol). Color degradation (L*, a*, and b* value) during thermal treatment was investigated, in particular as the “a*” value (the value of green color). Thermal degradation (thermal treatments between 55 and 80 °C per 90 min) of the green color of the leaves followed a fractional conversion model and the temperature dependence of the inactivation rate constant can be described using the Arrhenius law. The kinetic parameters using a reference temperature (T_refC = 70 °C) were determined as k_refC = 13.53 ± 0.01 × 10^?2 min^?1 and E_aC = 88.78 ± 3.21 kJ/mol. The results indicate that severe inactivation of POD (as an indicator for improved shelf life of the cooked vegetables) is accompanied by severe color degradation and that conventional cooking methods (typically 10 min/100 °C) lead to a high residual POD activity suggesting a limited shelf life of the cooked vegetables.     

Impact of Supercritical Carbon Dioxide and High Pressure on Lipoxygenase and Peroxidase Activity

The effects of supercritical carbon dioxide (ScCO2) treatment and high hydrostatic pressure treatment on the activities of lipoxygenase (LOX) and peroxidase (POD) were studied. Hydrostatic pressure treatment (240 MPa, 55 °C, 15 min) of LOX and POD in 30% sucrose solutions without buffer led to approximately 80% and approximately 50% residual activity, respectively. Application of ScCO2 (35.2 MPa, 40 °C, 15 min for LOX and 62.1 MPa, 55 °C, 15 min for POD) achieved approximately 35% LOX and approximately 65% POD inactivity in 30 % sucrose solutions. Total inactivation of LOX (10.3 MPa, 50 °C and 15 min) and of POD (62.1 MPa, 55 °C and 15 min) could be achieved through ScCO2 treatment of unbuffered solution. Increasing the concentration of sucrose and buffering (pH range 4 to 9) of enzyme solutions resulted in increased resistance of the enzymes to ScCO₂ treatment.     

高静压技术对豆浆中脂肪氧合酶的钝化效果及其动力学分析

研究了高静压技术(HHP)对豆浆中脂肪氧合酶(LOX)活性的钝化作用,并进行失活动力学分析。结果表明:HHP处理能显著钝化豆浆中LOX活性。用300～600MPa压力处理5～20min,对LOX活性有显著的钝化效果(P<0.05),并且随处理压力和处理时间的增加,酶的失活率提高。当压力500MPa,保压时间0min时也有一定的钝化LOX效果;当压力600MPa处理10min以上,豆浆LOX活性被完全抑制。用500MPa处理5min,可以达到与传统巴氏杀菌相同的钝酶效果(P>0.05)。HHP技术钝化豆浆LOX的过程可用一级动力学模型拟合(R2>0.900)。随着压力的升高和处理时间的延长,k值逐渐升高,D值逐渐减小;动力学参数ZP和Va分别为125.94MPa和-45.290cm3/mol。HHP技术在钝化豆浆中脂肪氧合酶活性方面比传统巴氏杀菌彻底,效果更好。其在改善豆浆品质方面具有良好的应用前景。     

脱脂豆粕中不同脂肪氧合酶活力对大豆分离蛋白凝胶性质的影响

通过加热处理低温脱脂豆粕，研究了豆粕中脂肪氧合酶的热失活动力学，并以此制备不同酶活的脱脂豆粕。分别提取分离蛋白。考察了不同分离蛋白的凝胶性质，并从蛋白溶液浊度、NSI值和碱溶上清液的分子量分布角度探讨了不同脂肪氧合酶活力对上述蛋白功能性质的影响。发现随豆粕中酶活力降低，所得分离蛋白的功能性质得到提高。     

Effects of Frozen Storage on Survival of Staphylococcus aureus and Enterotoxin Production in Precooked Tuna Meat

This study investigated the survival of Staphylococcus aureus in precooked tuna meat for producing canned products during frozen storage (?20 ± 2 °C) as well as its growth and enterotoxin production at 35 to 37 °C after the storage. Samples (50 ± 5 g) of precooked albacore (loin, chunk, and flake) and skipjack (chunk and flake) tuna were inoculated with 5 enterotoxin‐producing strains of S. aureus at a level of approximately 3.5 log CFU/g and individually packed in a vacuum bag after 3 h incubation at 35 to 37 °C. Vacuum‐packed samples were stored in a freezer (?20 ± 2 °C) for 4 wk. The frozen samples were then thawed in 37 °C circulating water for 2 h and incubated at 35 to 37 °C for 22 h. Populations of S. aureus in all precooked tuna samples decreased slightly (<0.7 log CFU/g) after 4 wk of storage at ?20 ± 2 °C, but increased rapidly once the samples were thawed and held at 35 to 37 °C. Total S. aureus counts in albacore and skipjack samples increased by greater than 3 log CFU/g after 6 and 8 h of exposure to 35 to 37 °C, respectively. All samples became spoiled after 10 h of exposure to 35 to 37 °C, while no enterotoxin was detected in any samples. However, enterotoxins were detected in albacore loin and other samples after 12 and 24 h of incubation at 35 to 37 °C, respectively. Frozen precooked tuna meat should be used for producing canned tuna within 6 to 8 h of thawing to avoid product spoilage and potential enterotoxin production by S. aureus in contaminated precooked tuna meat.     

Purification and Kinetic Characterization of Peroxidase from Tomato Cultivated under Different Salinity Conditions

By including hydrophobic chromatography in the purification scheme, 2 homogeneous tomato fruit peroxidase isoenzymes were obtained. The expression of these 2 peroxidases, one acid and the other basic, was determined in tomato fruits grown under different salinity conditions. Increased salinity modified the isoenzyme profile of tomato peroxidase. In tomatoes grown under highly saline conditions, there was an increase in the expression of the acid form with respect to the basic, the acid/basic ratio rising from 4.5 in tomatoes grown under normal saline conditions to 70 in those grown in highly saline conditions. Kinetic studies using 2,2'-azinobis(3-ethylbenzo-thiazolinesulfonic acid) as reducing substrate showed that increased salinity in the growth medium did not modify the kinetic parameter of tomato peroxidase over both hydrogen peroxide or reducing substrate.     

Reference Materials for Calibrating Probes Used for Measuring Thermal Conductivity of Frozen Foods

Reference materials which contained either 8% or 10% bentonite pastes and very fine pure copper powder (300 mesh, 30–.55%), provided thermal conductivities ranging from 0.9 to 1.9 W/mK. These materials were used to calibrate probes for measuring thermal conductivity of frozen food materials. Thermal conductivity values measured by the steady-state (parallel plate) method, the unsteady-state heat transfer (heating curve) method, and the probe method agreed within 5%. Although experimental density and heat capacity data of copper-filled bentonite pastes agreed with a mass average value of component properties, measured thermal conductivity values were much lower than the volume fraction average values.     

草莓冻藏过程中多酚氧化酶、过氧化物酶及维生素C的变化研究

本文研究速冻草莓在-12、-18、-40℃三种温度下十四周贮藏中可溶性多酚氧化酶（Polyphenol Oxidase，简称PPO）、过氧化物酶（Peroxidase，简称POD）及维生素C的变化，结果表明：不同温度下冻藏的草莓维生素C含量均呈下降趋势：冻藏初期可溶性多酚氧化酶、过氧化物酶活性上升，后期显著下降。