Inactivation of polyphenol oxidase from frozen red raspberry (Rubus idaeus L.) by high pressure carbon dioxide treatment

The effect of high pressure carbon dioxide (HPCD) treatment on polyphenol oxidase (PPO) from frozen red raspberry (Rubus idaeus L.) was evaluated. Moreover, the inactivation kinetics of its PPO was simulated by first‐order reaction theory. The minimum of PPO residual rate was 36.6% under 30 MPa and 55 °C for 60 min by HPCD treatment, while that was 66.8% at 55 °C for 60 min by thermal treatment. Moreover, the decimal reduction time of PPO decreased rapidly after HPCD treatment, compared to that of the thermal treatment. The thermal treatment at 55 °C takes a similar time to reach 10% PPO residual rate with HPCD treatment under 30 MPa at 35 °C. One reason for the results was that activation energy of PPO reduced from 98.9 to 14.6 kJ mol^?1 after HPCD treatment. Therefore, HPCD treatment showed stronger capacity to inactivate PPO from frozen red raspberry than the thermal treatment at same temperature.     

Inactivation of peroxidase and polyphenol oxidase in red beet (Beta vulgaris L.) extract with continuous high pressure carbon dioxide

The inactivation of peroxidase (POD) and polyphenol oxidase (PPO) in red beet extract (RBE) with continuous high pressure carbon dioxide (HPCD) was investigated. HPCD treatment at 7.5 MPa (55 °C, 30 min) resulted in a reduction of their activities by approximately 73% and 93%, respectively. Compared with thermal treatment, continuous HPCD treatment reduced the decimal reduction time (D) of POD and PPO from 555.6 min to 55.9 min and 161.3 min to 32.1 min, respectively. The inactivation process could be described by first-order kinetics (r² > 0.70, p < 0.05); D values declined when temperature increased and continuous HPCD at 7.5 MPa and 55 °C resulted in the highest reaction rate constant (k value; smallest D value). The activation energy of the inactivation was reduced by HPCD treatment from 92.5 kJ/mol to 69.8 kJ/mol and 57.1 kJ/mol to 49.5 kJ/mol for POD and PPO, respectively. Continuous HPCD treatment had little effect on the antioxidant capacities of RBE samples.     

Inactivation kinetics of apple polyphenol oxidase in different pressure–temperature domains

The impact of high hydrostatic pressure and temperature on the stability of polyphenol oxidase (PPO) was studied in cloudy apple juice. Application of 200–500 MPa near room temperature or heat treatment at 45–55 °C at ambient pressure caused an increase of PPO activity of up to 65% in freshly squeezed apple juice. Combined pressure–temperature inactivation experiments with fully activated PPO (5 min treatment at 400 MPa and 20 °C) were carried out in the range of 0.1–700 MPa and 20–80 °C. Enzyme inactivation kinetics followed a 2.2 order reaction scheme at all pressure–temperature conditions tested. A polynomial model was successfully applied to describe the rate of PPO inactivation as a function of pressure and temperature and was used to construct a pressure–temperature isokinetic diagram. This diagram clearly showed synergistic effects of pressure and temperature on the inactivation of apple PPO at pressures above 300 MPa and antagonistic effects at lower pressures. Compared to ambient pressure conditions, temperatures required to inactivate PPO in apple juice were increased 10–15 °C at 100–300 MPa.

Industrial relevance

High pressure processing of fresh fruits is gaining popularity in the food industry because of its ability to inactivate microorganisms and some enzymes near room temperature with little impact on flavour or nutritional attributes of the food. However, quantitative data regarding the impact of process parameters on the target reaction are required to economically utilise this technology. This paper provides a mathematical model describing the combined effect of pressure, temperature and treatment time on the inactivation of PPO in cloudy apple juice.     

Inactivation of polyphenol oxidases in cloudy apple juice exposed to supercritical carbon dioxide

The inactivation of polyphenol oxidase (PPO) in cloudy apple juice exposed to supercritical carbon dioxide (SCCO₂) treatment was investigated. Higher pressure, higher temperature, and longer treatment time caused more inactivation of PPO. The maximum reduction of PPO activity reached more than 60% at 30 MPa and 55 °C for 60 min. The experimental data followed first-order reaction kinetics; the kinetic rate constant k and the decimal reduction time D were closely related to the pressure and temperature of SCCO₂ treatment. Higher pressures or higher temperatures resulted in lower D values (higher k), the D value of PPO was minimized to 145 min treated by the combination of 30 MPa and 55 °C. Activation energy of 18.00 kJ /mol, was significantly reduced by SCCO₂ treatment at 30 MPa, as compared to activation energy of 72.0 kJ/mol for identical treatment at atmospheric pressure. Pressure and temperature sensitivity of kinetic parameters were studied. Z_P at 55 °C was 66.7 MPa and Z_T at 30 MPa was 108 °C.     

Comparison of the inactivation kinetics of pectin methylesterases from carrot and peach by high-pressure carbon dioxide

The inactivation of pectin methylesterases (PMEs) from carrot and peach in buffer by high-pressure carbon dioxide (HPCD) at 55 °C was investigated. The two PMEs were effectively inactivated by HPCD, their residual activity (RA) decreasing with increasing pressures. The RA of the two PMEs exhibited a fast decrease firstly and reached a constant after a prolonged treatment time; their inactivation kinetics was adequately modelled by a fractional-conversion model. The non-zero RA(A_∞)of the two PMEs was 6–7%, with increasing pressures the kinetic rate constant, k, increased and the decimal reduction time, D, decreased for the HPCD-labile fraction of the two PMEs. The labile fraction of carrot PME was more susceptible to HPCD than that of peach PME; the activation volume, V_a, and Z_P (the temperature increase needed for a 90% reduction of D) was −1079.37 cm³/mol and 5.80 MPa for carrot PME, and −130.51 cm³/mol and 48.31 MPa for peach PME.     

Inactivation kinetics of peroxidase and polyphenol oxidase in peach juice treated with gaseous ozone

The effectiveness of gaseous ozone for inactivating peroxidase (POD) and polyphenoloxidase (PPO) in peach juice was investigated. The suitability of first‐order and Weibull models to describe inactivation kinetics was also analysed. Peach juice was exposed to ozone (0.11 and 0.20 mg O₃ min^?1 mL^?1) in a bubble column up to 12 min at 20 ± 1 °C. Enzyme activities were reduced due to treatments. The magnitude of the inactivation increased with ozone level and exposure time. Reductions in activity after 12 min of treatment ranged between 99.5% and 99.8% for POD and between 93.9% and 97.3% for PPO, depending on ozone concentration. Inactivation curves were successfully fitted with the first‐order and Weibull models; although, based on the root‐mean‐square error, the corrected Akaike and the Bayesian Schwarz criterion, the Weibull model showed stronger capability in all cases.     

超高压技术在鲜榨桃汁加工中的应用

以鲜榨桃汁为原料,分别研究了热处理对其多酚氧化酶(PPO)、过氧化物酶(POD)的灭酶效果影响和超高压对其微生物的灭菌效果影响。结果显示:PPO、POD相对残存酶活随着热处理温度和时间的增加而减小,PPO对热处理的耐受性明显高于POD,最适灭酶条件为75℃1min;菌落总数、霉酵总数和乳酸菌数均随着超高压处理压力和时间的增加而减少,霉菌和酵母菌对超高压的耐受能力明显差于细菌,最适灭菌条件为600MPa10min;按以上最适条件生产的桃汁在1个月贮藏期内达到商业无菌。     

Inactivation of polyphenol oxidase from Pacific white shrimp by dense phase carbon dioxide

The inactivation of polyphenol oxidase (PPO) from Pacific white shrimp exposed to dense phase carbon dioxide (DPCD) treatment was investigated. PPO activity showed a dramatic loss at 4–25 MPa and 37 °C. At the lower pressure (4–15 MPa), the experimental data of inactivation followed the first-order reaction kinetic model, the pressure sensitivity (Z_P) of the kinetic parameters was 49.02 MPa and the activation volume (△ V^≠) was − 120.88 cm³/mol. At the higher pressure (20 and 25 MPa), the experimental data of inactivation followed the biexponential kinetic model. The kinetic rate constant k_F and k_S of fast and stable fractions were 2.45 and 0.08 min^{− 1}, respectively. The decimal reduction time D_F and D_S were 0.94 and 29.43 min at 25 MPa and 37 °C, respectively. After DPCD treatment, the loss activity of PPO had no restoration storing for 6 days at 4 °C. The results of SDS-PAGE and activity staining also showed that DPCD treatment had the obvious inhibitory effect on PPO from Pacific white shrimp. The PPO activity in vivo was easier to be inactivated than that in crude PPO extracts under the same DPCD treatment conditions.

Industrial relevance

There is a growing interest in non-thermal pasteurization methods, which could retain food's freshlike physical, nutritional, and sensory qualities. Pacific white shrimp accounts for 90% of the global aquaculture shrimp production, they are becoming increasingly popular. However, enzymatic browning of shrimp has been of great concern to food scientists and food processors. Dense phase carbon dioxide (DPCD) may be an adequate tool to obtain high quality since PPO activity could not be inactivated totally by high pressure under 400 MPa yet. The present work deals with the inactivation of PPO from Pacific white shrimp exposed to DPCD treatment in order to explore the feasibility of shrimp by DPCD process.     

Enzyme Activity and Nutritional Quality of Peach (Prunus persica) Juice: Effect of High Hydrostatic Pressure

The inactivation of polyphenol oxidase and pectin methylesterase in peach juice was investigated after high hydrostatic pressure processing at 400–600 MPa and 25°C for 5–25 min, respectively. At 400 MPa, polyphenol oxidase and pectin methylesterase were activated by 7.3 and 2.6%. At 500 and 600 MPa, polyphenol oxidase and pectin methylesterase were inactivated significantly with increasing the pressure and time, and the inactivation kinetics was fitted by the first order model. Moreover, some physio-chemical properties were studied. The results revealed that high hydrostatic pressure treatment preserved more L-ascorbic acid and maintained the color and sensory quality better than thermal treatment.     

Inactivation Kinetics of the Most Baro-Resistant Enzyme in High Pressure Processed Litchi-Based Mixed Fruit Beverage

This work focused on a litchi-based mixed fruit beverage, comprising of coconut water and lemon juice, mixed in an optimized proportion. Based on preliminary studies, three resistant spoilage enzymes were identified in the beverage, viz. polyphenol oxidase (PPO), peroxidase (POD), and pectin methyl esterase (PME). The response surface methodology (RSM) based on central composite face-centered design (FCCD) screened out PPO as the most resistant enzyme within the high pressure processing (HPP) domain of 200–600 MPa/30–70 °C/0–20 min. A detailed kinetic study was conducted on PPO inactivation within the same HPP domain along with a set of thermal treatments (0.1 MPa/30–70 °C). A synergistic effect of pressure and temperature on PPO inactivation was observed, throughout the HPP domain. However, PPO was almost completely inactivated at 500 MPa/70 °C/20 min. The inactivation order (n) values for PPO were 1.10 and 1.25 for thermal and HPP treatments, respectively. For every 10 °C rise in temperature, the inactivation rate constant (k, U^n-1 min^?1) increased approximately by 1.5 times, within 50–70 °C (at 0.1 MPa), while a 10-fold increase was obtained in the case of HPP treatments. The activation energy (E _a ) and the activation volume (V _a), depicting the temperature and pressure dependence of k, was found to decrease slightly, with an increase in pressure and temperature, respectively. The PPO inactivation rate constant was modeled as a function of both temperature and pressure conditions by combining both Arrhenius and Eyring equations.     

The contribution of high pressure carbon dioxide in the inactivation kinetics and structural alteration of myrosinase

High pressure carbon dioxide (HPCD) has been verified to be an efficient way of inactivating enzyme activity. This work investigates the influence of temperature (T), pressure (P), exposure times (t) on the activity of commercial myrosinase (MYR) submitted to HPCD. Results showed that only 1.00% of MYR activity retained at 22 MPa and 65 °C for 5 min. Moreover, the first‐order reaction kinetic data of MYR inactivation as influenced by pressure of HPCD were analysed. With the pressure rising from 8 to 22 MPa at 55 °C, the inactivation rate constant (k) increased from 0.015 to 0.024 min^?1, while the decimal reduction time (D) decreased from 157.2 to 96.1 min. Additionally, a series of exploratory experiments were conducted to investigate the contribution of the HPCD parameters (T, P and CO₂ dissolution), with analysing circular dichroism spectroscopy and tryptophan fluorescence spectra, illustrate that CO₂ dissolution plays a dominant role in MYR inactivation and structural alteration.     

Kinetic study of high pressure microbial and enzyme inactivation and selection of pasteurisation conditions for Valencia Orange Juice

Keeping quality of fresh orange juice is highly dependent on pectinolytic enzyme activity and the growth of spoilage microorganisms. The inactivation kinetics of indigenous pectin methylesterase (PME) and of the two more pressure resistant species of spoilage lactic acid bacteria (LAB) Lactobacillus plantarum and L. brevis in freshly squeezed Valencia orange juice under high hydrostatic pressure (100–500 MPa) combined with moderate temperature (20–40 °C) was investigated. PME inactivation followed first order kinetics with a residual PME activity (15%) at all pressure–temperature combinations used. The values of activation energy and activation volume were estimated at each pressure and at each temperature, respectively. Values of 90 kJ/mol and ?30 mL/mol at reference pressure of 300 MPa and reference temperature of 35 °C were estimated respectively. The corresponding z_T and z_P values of LAB inactivation were also estimated at all conditions tested. Values of 19.5 °C and 95 MPa at reference pressure of 300 MPa and reference temperature of 30 °C were estimated respectively for L. plantarum, while the corresponding values for L. brevis were 40 °C and 82 MPa, respectively, at the same reference conditions. Pressure and temperature were found to act synergistically both for PME and LAB inactivation. The PME and LAB inactivation rate constants were expressed as functions of the temperature and pressure process conditions. These functions allow the determination of the pressure/temperature conditions that achieve the target enzyme and microbial inactivation at a selected processing time. The process conditions of 350 MPa at 35 °C for 2 min are proposed as effective for Valencia orange juice cold pasteurisation.     

Change of polyphenol oxidase activity, color, and browning degree during storage of cloudy apple juice treated by supercritical carbon dioxide

The effects of supercritical carbon dioxide (SCCO₂) treatment of 8, 15, 22, and 30 MPa for 60 min at 55 °C on polyphenol oxidase (PPO) activity, color, and browning degree in cloudy apple juice during storage at 4 °C for 4-weeks were investigated. The SCCO₂ treatment had significant effects on inactivation of PPO and the least residual activity of PPO was 38.50% at 30 MPa. The restoration of PPO residual activity after SCCO₂ treatment was also observed, which was dependent on the pressure level. A greater reduction of lightness L and a minor increase of redness a of cloudy apple juices after SCCO₂ treatment occurred. Moreover, the total color difference (ΔE), which was significantly less than that of untreated sample, was decreased by enhancing the pressure level. The changes of lightness L and browning degree A during storage were well fitted to a first-order kinetic model. The rate constants of k _L and k _A of samples subjected to SCCO₂ treatment reduced from 4.75×10⁻² to 0.42×10⁻² and 37.19×10⁻² to 8.02×10⁻², respectively, when pressure increased from 0 MPa (untreated sample) to 30 MPa.     

Inactivation of Escherichia coli by high hydrostatic pressure at different temperatures in buffer and carrot juice

The inactivation of Escherichia coli MG1655 was studied at 256 different pressure (150-600 MPa)-temperature (5-45 degrees C) combinations under isobaric and isothermal conditions in Hepes-KOH buffer (10 mM, pH 7.0) and in fresh carrot juice. A linear relationship was found between the log10 of inactivation and holding time for all pressure-temperature combinations in carrot juice, with R2-values>or=0.91. Decimal reduction times (D-values), calculated for each pressure-temperature combination, decreased with pressure at constant temperature and with temperature at constant pressure. Further, a linear relationship was found between log10D and pressure and temperature. A first order kinetic model, describing log10D in carrot juice as a function of pressure and temperature was formulated that allows to identify process conditions (pressure, temperature, holding time) resulting in a desired level of inactivation of E. coli. For Hepes-KOH buffer, the Weibull model more accurately described the entire set of inactivation curves of E. coli MG1655 compared to the log-linear or the biphasic model. Several secondary models (first and second order polynomial and Weibull) were evaluated, but all had poor fitting capacities. When the Hepes-KOH dataset was limited to 22 of the 34 pressure-temperature combinations, a first order model was appropriate and enabled us to use the same model structure as for carrot juice, for comparative purposes. The major difference in kinetic behaviour of E. coli in buffer and in carrot juice was that inactivation rate as a function of temperature showed a minimum around 20-30 degrees C in buffer, whereas it increased with temperature over the entire studied temperature range in carrot juice.     

High Pressure Inactivation of Polyphenoloxidases

Pressure stabilities of polyphenoloxidases (PPO) from apples, avocados, grapes, pears and plums were determined at pH 6-7. These PPOs differed in pressure stability, but all were rather pressure-stable. Inactivation of PPO from apple, grape, avocado and pear at room temperature (25°C) became noticeable at 600, 700, 800 and 900 MPa respectively, and followed first-order kinetics. Plum PPO was not inactivated at room temperature by pressures up to 900 MPa. For the two most pressure-stable PPOs, we investigated whether pressure stability would be reduced by the simultaneous application of mild heat. In case of plum PPO, activity reduction was detectable at 900 MPa and 50°C. Further temperature increase resulted in increase of the inactivation rate constant (E_a 63 kJ/mol). In case of pear PPO, temperature increase up to 35°C resulted in a 3-fold reduction of the inactivation rate constant. Only at higher temperatures, increase of the inactivation rate constant with increasing temperature was noted (Ea 120 kJ/mol).     

高静压对桃汁杀菌、钝化酶活性的效果

研究在不同处理压力和时间条件下,高静压加工技术对桃汁中微生物(细菌总数、霉菌、酵母菌、大肠菌群)以及酶(多酚氧化酶、果胶甲基酯酶、脂肪氧化酶)的影响。结果表明:经400MPa、5min高静压处理即可完全杀灭桃汁中的微生物;在400MPa和500MPa条件下,桃汁中的多酚氧化酶和脂肪氧化酶的活性出现了不同程度的激活现象,但在600MPa时,随着处理时间的延长,其活性逐渐降低,经30min处理后,分别被钝化了0.7662和0.641。而果胶甲基酯酶在400、500、600MPa条件下,出现了不规律的激活或钝化现象。另外,研究表明在高静压加工前增加漂烫工艺,可以有效杀灭桃汁中的微生物及钝化酶活性。     

超高静压对非浓缩还原杨梅果汁中氧化酶的钝化作用

该研究以非浓缩还原（not from concentrate,NFC）杨梅果汁为研究对象,研究不同超高静压（high hydrostatic pressure,HHP）处理（300~600 MPa/0~30 min）对NFC杨梅汁中多酚氧化酶（polyphenol oxidase,PPO）和过氧化物酶（peroxidase,POD）的影响。对比传统高温灭酶,拟合建立HHP压力与酶活性的一级动力学回归方程,分析得到相关参数（压力脉冲效应PE、压力脉冲数值ND、等效破坏值Dp及酶的失活速率K）。结果表明,较高压力（400~600 MPa）对PPO与POD均起到钝化效果,其中600 MPa/10 min能钝化90%的PPO活性,600 MPa/20 min钝化80%的POD活性。600 MPa/30 min条件下,重复加压不能明显加强钝化效果。将PPO和PPO活性与压力进行一级动力学拟合,得到相应线性回归方程（R2>0.8）。随着压力从300 MPa升高到600 MPa,PPO的K值从3.03×10-2升高到12.12×10-2,POD的K值从1.23×10-3上升到7.67×10-3。600 MPa条件下,PPO和POD的ND分别为1.04和1.59,Dp值都为19。同时,压力和保压时间及其相互作用对PPO和POD活性的影响均有极高的显著性（p<0.001）。因此,HHP对杨梅果汁中关键的氧化酶能起到较好的钝化作用,能够为NFC杨梅汁加工技术的应用提供科学依据。     

Thermal inactivation kinetics parameters of browning enzymes in starfruit (Averrhoa carambola L.) juice

This study aimed to evaluate the thermal inactivation kinetics of polyphenol oxidase (PPO) and peroxidase (POD) in starfruit juice. It followed the Malaysia Food Regulations 1985 and CODEX STAN 247-2005. Glucose, fructose and sucrose were the main sugars in starfruit juice. The total soluble solids, pH, titratable acidity, and total phenolics content of the starfruit juice produced were 8.13 ± 0.25 °Brix, 3.80 ± 0.05, 0.43% ± 0.02% malic acid, and 93.67 ± 4.96 mg GAEL⁻¹, respectively. Thermal inactivation kinetics of PPO and POD followed the first-order kinetic model. The decimal reduction time at 83.6 °C (D_83.6) of PPO and POD was 198.48 and 98.4 s, respectively, while the thermal resistance constant (z value) of PPO and POD was 12.8 and 5.4 °C, respectively. In conclusion, PPO might be a suitable signal for thermal processing on starfruit juice since it has higher heat resistance than POD.     

High pressure and thermal inactivation kinetics of polyphenol oxidase and peroxidase in strawberry puree

The objective of this work was to study the thermal and high pressure inactivation kinetics of polyphenol oxidase (PPO) and peroxidase (POD) in strawberry puree. PPO from two strawberry cultivars (‘Festival’ and ‘Aroma’) was found to be highly thermostable in strawberry puree with no significant inactivation even after 30 min treatment at 100 °C. In contrast, POD from the two cultivars displayed very high thermosensitivity with complete inactivation in less than 5 min at 70 °C. The thermal inactivation kinetics of strawberry POD was described by a biphasic model. The activation energies for the inactivation of the stable and the labile fractions were estimated to be 254.9 and 221.6 kJ/mol respectively. Combined high pressure–thermal treatment at pressures ranging from 100 to 690 MPa, temperatures ranging from 24 to 90 °C and treatment times between 5 and 15 min did not have significant effect on PPO while substantial inactivation of POD was observed. The inactivation kinetics of POD during combined high pressure–thermal processing was well described by first-order kinetics probably due to the inactivation of the labile fraction during the pre-heating and the compression phase.Industrial relevanceThe oxidative enzymes polyphenol oxidase and peroxidase cause the degradation of anthocyanins and other polyphenols in strawberry products, leading to discoloration and loss of antioxidant activity. In this work the thermal and high pressure inactivation of strawberry polyphenol oxidase and peroxidase was investigated so as to assess the suitability of high pressure processing as an alternative to thermal processing. Strawberry polyphenol oxidase was found to be highly resistant to both thermal and high pressure inactivation. Thus in order to maintain the quality of processed strawberry products, high pressure processing should be accompanied by additional measures such as exclusion of oxygen, refrigerated storage and the use of natural enzyme inhibitors.     

高压均质对芒果汁中大肠杆菌的杀菌动力学

为预测高压均质(high pressure homogenization,HPH)对芒果汁中大肠杆菌的杀菌作用,进行了压力40~190 MPa、进料温度20~60℃和均质次数1~5次的HPH实验,并运用Weibull模型对杀菌致死曲线进行动力学分析。研究表明:随压力、进料温度和均质次数的升高,HPH对芒果汁中大肠杆菌的杀菌效果增强。50℃HPH处理芒果汁,压力由40 MPa升高至190 MPa时,大肠杆菌降低量由0.46(lg(CFU/mL))增加至5.16(lg(CFU/mL)),达到美国食品药物管理局规定的非热加工杀菌卫生安全要求;70 MPa HPH处理芒果汁,进料温度由20℃升高至60℃时,大肠杆菌降低量由0.34(lg(CFU/mL))增加至5.02(lg(CFU/mL));20℃、190 MPa HPH处理芒果汁,均质次数由1增加至4时,大肠杆菌降低量由1.73(lg(CFU/mL))增加至5.15(lg(CFU/mL))。通过Weibull模型拟合杀菌致死曲线,并对模型进行简化,发现简化的Weib ull模型在20~50℃、40~190 MPa均质1次和20~40℃、190 MPa均质1~5次时拟合性较好(R~20.92)。简化的Weibull模型可用于预测进料温度-压力和进料温度-均质次数的杀菌效果,可为芒果汁的HPH生产过程中微生物安全性的控制提供理论依据。