Enzyme Inactivation Kinetics Research Articles

Pulsed light technology for fresh‐cut produce: A review on mechanism and inactivation kinetics of microbes and enzymes

AbstractConsumers are becoming more aware of the benefits of eating healthy foods like fresh‐cut produce without compromising the quality of sensory and nutritional aspects. Emerging non‐thermal technology like pulsed light (PL) is one of the most potential surface decontamination methods for fresh‐cut produce. Unlike conventional methods, PL does not cause unwanted changes in finished products like nutritional losses, color, textural changes, production of off‐flavors, etc. Its application is confined to not only surface disinfection but also enzyme inactivation. The death of microbes is due to PL's photochemical and photothermal effects. The photochemical effect is also responsible for enzyme inactivation, and Weibull or log‐linear models can explain their inactivation kinetics. Despite several advantages, PL has limitations of less effectiveness in eliminating microorganisms due to low penetration into the tissue of fresh‐cut produce, which can be overcome by applying different absorption‐enhancing agents to improve absorption capacity. Moreover, in‐package treatment can avoid the problem of recontamination of the treated products. This review provides an overview of the mechanisms and kinetics involved in the inactivation of microbes and enzymes in PL treatment. Additionally, this paper reviews the advantages and limitations of utilizing PL technology for shelf‐life enhancement of fresh‐cut produce and compiles the recent works done to enhance the efficacy of PL treatment. Furthermore, future scope and remedies to solve the existing problems of PL and for commercialization of this technique in the field of fresh produce are discussed.

Inactivation kinetics of polyphenol oxidase and peroxidase enzymes in buffer solutions by ultraviolet light sources at multiple UV-C wavelengths

This study aimed to evaluate the effect of multiple wavelengths of UV-C light sources on the inactivation of polyphenol oxidase (PPO) and peroxidase (POD) in buffer solutions. Enzyme solutions were treated with a UV-C excimer lamp (EL) emitting at 222 nm, low-pressure mercury (LPM) lamp at 253.7 nm, four UV-LEDs sources emitting at 255, 265, 275 and 285 nm, and a novel pulsed UV-C lamp (PL) emitting at 232, 242, 261 and 272 nm. Three kinetic models were used to compare UV-C wavelengths enzymatic inactivation efficacy considering the volume average fluence for 90% reduction (HD). Validation of kinetic parameters was performed under continuous-flow treatment in a UV-C LED and EL system. EL treatment resulted in the highest inactivation efficacy of PPO and POD enzymes with HD of 52.8 and 74.8 mJ/cm2, respectively. PL inactivation efficacy for both enzymes was higher than with LPM lamp or UV-LEDs. The two-component first-order model best described the inactivation of PPO and POD in a buffer solution. The adjusted kinetic parameters for this model were validated under UV-C light continuous flow processing. This study contributes to the development of liquid food treatment by ultraviolet technology.

Modeling the inactivation kinetics of polyphenol oxidase and peroxidase during the cold plasma treatment of kiwifruit juice

AbstractCold Plasma (CP) is an emerging non‐thermal technology that preserves and decontaminates food. Kiwifruit juice spoilage enzymes polyphenol oxidase (PPO) and peroxidase (POD) were studied at 24‐36 kV, 2‐10 mm juice depth, and 2‐10 min treatment time using log‐linear, Weibull distribution, and logistic models. The Weibull and logistic model explained enzyme inactivation kinetics well based on goodness of fit (Coefficient of determination: 0.97‐0.99 & root mean square error: 0.11‐2.15) and validation indices (accuracy factor: 1‐1.06 and bias factor: 0.96‐1.04). The logistic model suited PPO and POD inactivation curves well, with the least Akaike weight in 71.29% and 61.45%, respectively. POD had a half‐maximal activity of 11.37, 5.22, and 3.60 min at 24 kV, 30 kV, and 36 kV at 6 mm juice depth, compared to 8.71, 4.41, and 2.51 min for PPO. This shows that POD is more CP‐resistant than PPO, making its inactivation in kiwifruit juice crucial.Practical ApplicationKinetic modeling helps to comprehend the fundamental process and is suitable for improving the inactivation process and enhancing its efficacy. As a relatively primary method, mathematical models facilitate a greater comprehension of the mechanism and factors influencing the inactivation processes. This study involves the inactivation of PPO and POD to produce a stable kiwifruit juice. The inactivation of PPO and POD enzymes will prevent the undesirable enzymatic browning of juice. The kinetic data related to enzyme inactivation will help in deciding the process parameters like plasma voltage, juice depth, and treatment time for producing enzymatically stable kiwifruit juice.

Effect of low intensity direct-current electric field on anti-browning of apple juice: Enzyme inactivation kinetics and juice properties

For the first time, the efficacy of low intensity (1–7 V/cm) direct-current (DC) electric field in preventing browning in apple juice is examined. The results show that applying a 5 V/cm electric field can inactivate 89.24% of polyphenol oxidase (PPO) within 90 min and achieve complete inactivation of peroxidase (POD) within 70 min, maintaining the juice temperature below 30 °C throughout the process. The Weibull model fits the PPO and POD inactivation effectively, suggesting varied responses among isoenzymes. Under the optimal conditions (5 V/cm for 90 min), a 15.89% increase in total phenolic content (TPC) was observed alongside significant improvements in antioxidant activity. Furthermore, this method preserved the natural flavor and lightened the color of the apple juice, enhancing its sensory appeal. These findings indicate that low intensity DC electric field is an innovative and effective choice for preventing juice browning, offering significant potential for application in the food industry.

Pulsed light treatment of table grape juice: Influence of matrix pH on microbial and enzyme inactivation kinetics

The influence of pulsed light (PL) treatment and matrix pH was tested in the case of green table grape juice. Escherichia coli, Listeria monocytogenes, and Saccharomyces cerevisiae were inoculated into the table grape juices with matrix pH levels of 3.0, 3.5, and 4.0. Further, the juice was treated at 378–3186 J/cm2. The Weibull model better explained the microbial inactivation data. The shape parameters (β) from the Weibull model for E. coli, L. monocytogenes, and S. cerevisiae are 0.95 ± 0.02, 0.55 ± 0.04, and 0.40 ± 0.03, respectively. A lesser δ-value (scale parameter in J/cm2) was obtained for each microorganism at a lower pH. PL exposure resulted in a lack of cell visibility, cell wall, and cell membrane disruption in bacterial cells. The inactivation of polyphenol oxidase (PPO), peroxidase (POD), and pectin methyl esterase (PME) was studied at an elevated fluence level of 1152–3186 J/cm2. Enzymes in the juice were less sensitive to PL than microorganisms. The enzyme inactivation of >90% was achieved at 3186 J/cm2; thus, the juice was adjudged as the microbially safe and enzymatically stable pasteurized sample. The PL-induced enzyme inactivation followed nth-order kinetics with an inactivation order (n) of 1.1, 1.15, and 0.85 for PPO, POD, and PME, respectively. The enzyme inactivation rate was higher at a low pH. The PL-treated pasteurized juice at pH 3.5 showed the highest sensory acceptance (7.4 out of 9), with a minimal color change, a 7% loss in total phenolics, and a 12% loss in vitamin C and flavonoids.

Pasteurization of tender coconut water by pulsed light treatment: Microbial safety, enzymatic inactivation, and impact on physicochemical properties

The study investigated the potential of pulsed light (PL) in the pasteurization of tender coconut water (TCW). The initial counts of E. coli, B. cereus and L. monocytogenes in the inoculated TCW were 7.00, 9.14 and 7.8 log10 cfu mL−1, respectively. For a PL fluence of 465 J cm−2, E. coli, B.cereus and L. monocytogenes exhibited a log reduction of 5.12, 2.97 and 3.40, respectively. Bacillus cereus and Listeria monocytogenes exhibited greater resistance than Escherichia coli in the TCW. Peroxidase (POD) was more sensitive to PL treatments than polyphenoloxidase (PPO) in TCW. Weibull model and nth order model exhibited excellent fit for microbial inactivation (R2 > 0.96) and enzyme inactivation (R2 > 0.97) kinetics, respectively. While 5-log10 reduction of B. cereus and L. monocytogenes was achieved at 2.5 kV|2.5 min (1073 J cm−2), PPO was inactivated by greater than 99% at 2.9 kV| 5 min (2988 J cm−2). While the total reducing sugars increased, the changes in color (0.49 < ΔE* < 1.51), pH, total soluble solids, and acidity were insignificant after the PL pasteurization. The PL condition of 2.9 kV|5 min preserved 21 and 24% more phenolics and ascorbic acid in TCW, along with greater sensory scores than the thermal treatment (90 °C|3 min). Industrial significanceThe outcome of this study determined the intensity (fluence of 2988 J/cm2) and penetration depth (4–5 mm) required for the pasteurization of tender coconut water (TCW). On an industrial scale of large processing volumes, continuous pulsed light (PL) pasteurization of TCW can be undertaken in an annular flow reactor or thin film flat bed chamber. The thickness of the film can be mimicked from this study to ensure adequate penetration of PL in the sample to achieve adequate lethality.

Impact of high‐pressure treatments on enzyme activity of fruit‐based beverages: an overview

SummaryWith an ever‐increasing demand for clean label products, there is a greater need for efficient and environmentally friendly processes to compete the conventional thermal or chemical treatments. For instance, high‐pressure processing (HPP) has been widely studied in the fruit industry from the last two decades. HPP can inactivate or activate different enzymes in fruit juices, pulp, and purées. HPP treatment inactivates the enzymes by the alterating the conformation of the protein structure and the active site. Depending on the enzyme, pressure, pH, temperature and treatment time, HPP can increase enzyme activity due to the release of membrane‐bound enzymes and also due to changes in protein conformation and active site that facilitate interaction with the substrate. Furthermore, the combination of high pressure, temperature and reduced treatment times offered greater inactivation of enzymes in fruit beverages. This study aimed to investigate the inactivation kinetics of endogenous enzymes in fruit beverages.

Phytochemical analysis, antioxidant and in vitro \u03b2-galactosidase inhibition activities of Juniperus phoenicea and Calicotome villosa methanolic extracts

BackgroundJuniperus Phoenicea (JP) and Calicotome Villosa (CV) are used by Jordanian populations as herbal remedies in traditional medicine. Herein, the phytochemical contents of their methanolic extracts were analyzed and their antioxidant as well as in vitro anti- β-Galactosidase activities were evaluated; their effect on β-Galactosidase enzyme kinetics was evaluated and the thermodynamic of the enzyme was determined.MethodsThe antioxidant activity of JP and CV crude methanolic extracts was evaluated using 1,1-diphenyl,2-picrylhydrazyl (DPPH) free radical scavenging and ferric reducing antioxidant power (FRAP) assays; however, the effect of the plants’ crude extracts on β-Galactosidase activity and kinetics was evaluated in vitro. Moreover, total phenolic, flavonoids, and flavonols content in plants’ extracts were determined and expressed in Gallic acid equivalent (mg GAE/g dry extract) or rutin equivalent (mg RE/g dry extract).ResultsPhytochemical screening of the crude extracts of JP and CV leaves revealed the presence of phenols, alkaloids, flavonoids, terpenoids, anthraquinones, and glycosides. Flavonoids and flavonols contents were significantly higher in JP than in CV (p < 0.05). Furthermore, an analogous phenolic content was detected in both JP and CV methanolic extracts (103.6 vs 99.1 mg GAE/g extract). The ability of JP extract to scavenge DPPH radicals was significantly higher than that of CV extract with IC50 = 11.1 μg/ml and 15.6 μg/ml, respectively. However, their extracts revealed relatively similar antioxidant capacities in FRAP assay; their activity was concentration dependent. The JP extract inhibited β—galactosidase enzyme activity with a significant IC50 value compared to CV extract; they exhibited their inhibitory activities at IC50 values 65 µg/ml and 700 µg/ml, respectively. Rutin revealed anti-β-galactosidase activity at IC50 = 75 µg/ml. The mode of inhibition of β-galactosidase by JP, CV, and rutin was non-competitive, mixed, and competitive inhibition, respectively. Thermodynamic and enzyme inactivation kinetics revealed that β-galactosidase has a half-life time of 108 min at 55 °C, activation energy of 208.88 kJ mol−1 and the inactivation kinetics follows a first-order reaction with k-values 0.0023–0.0862 min−1 and positive entropy of inactivation (∆S°) values at various temperatures, indicating non-significant processes of aggregation.ConclusionsThe methanolic extracts of JP and CV possess anti-hyperglycemic and antioxidant activities with potential pharmaceutical applications.

Effect of UV-C irradiation on the inactivation kinetics of oxidative enzymes, essential amino acids and sensory properties of coconut water.

The impact of ultraviolet light (UV-C) irradiation on oxidative enzymes [Polyphenol oxidase (PPO) and Peroxidase (POD)], free essential amino acids and sensory profile of coconut water were investigated. PPO and POD activities were lost to 94 and 93%, respectively of its original value at fluence level of 400mJ/cm2. Inactivation kinetics of both enzymes were fitted to nonlinear Weibull model with an increase in UV dosage with a high coefficient of determination (R2 > 0.97) and low root mean square error (RMSE < 0.06). No significant change was observed in all essential amino acids (p > 0.05) after UV-C treatment up to maximum delivered fluence of 400mJ/cm2. Sensory attributes of coconut water up to a treated UV-C fluence level of 200mJ/cm2 were well retained in terms of chosen descriptors (p > 0.05). This study allow to further investigate the development of UV-C light technology for inhibition of spoilage enzymes and prolonged shelf-life of low acid beverages.

Thermal inactivation kinetics and effects of drying methods on the phenolic profile and antioxidant activities of chicory (Cichorium intybus L.) leaves

The thermal inactivation kinetics of enzymes, including polyphenol oxidase (PPO) and peroxidase (POD), in chicory (Cichorium intybus L.) leaves were evaluated. In addition, the influences of different drying techniques (shade drying, hot air drying and freeze drying) on the phenolic profiles and antioxidant activities of chicory leaves were determined. The antioxidant activities of chicory leaves were evaluated on the basis of their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, reducing power, and 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. The results showed that the activation energy for PPO and POD inactivation were 123.00 kJ/mol and 78.99 kJ/mol, respectively. Preliminary treatment with hot water for 3 min at 90 °C was beneficial for preserving the phenolics present in fresh leaves. Hot air drying was better for the phenolics preservation. The hot air-dried and freeze-dried leaves possessed good antioxidant activities. The leaves with higher phenolics contents had better antioxidant activities, which indicated that the preservation of the phenolics was important for maintaining the antioxidant activity of chicory leaves.

Drying characteristic, enzyme inactivation and browning pigmentation kinetics of controlled humidity-convective drying of banana slices

Investigating the kinetics of enzyme activities and browning indexes in food are very essential in understanding the enzyme inactivation and browning pigmentation reaction during drying processing. In order to understand and predict accurately the enzyme inactivation and browning pigmentation of banana slices using Relative Humidity (RH)-convective hot air dryer aided by ultrasound (US) pretreatment, this study was conducted. Drying was carried out with 20 kHz frequency of US-pretreatment using three durations (10 20 and 30 min) and RH (10 20 and 30%) conditions at 70 °C and 2.0 m/s air velocity. The kinetic study of both enzyme inactivation and browning pigmentation results were compared to their relevance of fit in terms of coefficient of correlation (R2), the root mean square error (RMSE) and the reduced chi-square (χ2). First order and second-order polynomial kinetic model fitted well for enzyme inactivation and browning indexes respectively. Both enzymes inactivation kinetics and enzymatic browning index (EBI) declined significantly (p < 0.05) with increasing drying time in all drying conditions and rate of decrease intensified in longer US-pretreatment duration and lower RH conditions. However, shorter US-pretreatment duration and higher RH conditions reduced the non- enzymatic browning index (NBI) significantly. Again, longer US-pretreatment duration and lower RH shortened the drying time but adversely created more microspores from the micrograph study. Longer US pretreatment and lower RH decrease significantly (p < 0.05) the L* and b* values whereas the a* values was increased.

Effect of thermosonication batch treatment on enzyme inactivation kinetics and other quality parameters of cloudy apple juice

Abstract Cloudy apple juice has been treated by thermosonication in batch mode as an alternative processing to thermal treatment. Thermosonication was found to be effective to inactivate polyphenol oxidase; however, pectinmethylesterase was found to be more resistant. An increase of the working ultrasound amplitude and the amount of energy supplied to the juice led to lower enzyme residual activities. Enzyme inactivation kinetics were determined at different temperatures (from 44 to 67 °C). Inactivation data were described by the first order kinetic model and the Weibull model, both models yielding good fitting. Thermosonication treatment caused a homogenization effect reflected in the shifting of the particle size distribution towards smaller diameters. The effect of the nature of dissolved gases in the juice on enzyme inactivation was studied by displacing the air dissolved in the juice by bubbling nitrogen or carbon dioxide, previous to the thermosonication experiments. Higher inactivation rates were obtained by displacing the air with nitrogen. Industrial relevance Consumers demand of natural and fresh-like products has driven the food industry to investigate alternative technologies to replace conventional food heat treatments that may affect food quality. Among these technologies, thermosonication treatment is an attractive technology that can inactivate microorganisms and enzymes. This work shows that some enzymes that cause deleterious effect on cloudy apple juice can be more effectively inactivated by thermosonication than by thermal treatment, in the same temperature range, reducing the damages caused by heating.

Evaluation of HPCD batch treatments on enzyme inactivation kinetics and selected quality characteristics of cloudy juice from Golden delicious apples

Cloudy apple juice has been treated by high pressure carbon dioxide (HPCD) as non-thermal technology to inactive polyphenoloxidase and pectinmethylesterase in batch mode. Stirring speed (from 200 to 600 rpm) induced an increase in the enzyme inactivation rate while a triple cycle of pressurization/depressurization led to the same enzyme inactivation efficiency. Enzyme inactivation kinetics were determined at different temperatures (from 35 to 45 °C) and pressures (from 10 to 20 MPa). Data were described by the first order kinetic model and the Weibull model. For the first order kinetic model, decimal reduction time for HPCD treatment was found to be smaller than for mild heating, in the same temperature range. The same tendency was observed for the first decimal reduction time in the Weibull model. HPCD treatment resulted in a homogenization effect reflected in the shifting of the particle size distribution towards smaller diameters after treatment. HPCD treatment did not result in a change of water and oxalate soluble pectin content, total phenolic compounds and hidroxymethylfurfural content.

Kinetic modeling of polyphenoloxidase inactivation during thermal-assisted high pressure processing in black tiger shrimp (Penaeus monodon)

High pressure inactivation kinetics of polyphenol oxidase (PPO) was studied in black tiger shrimp (Penaeus monodon), with the applied processing parameters ranging from 300 to 600 MPa, 30–60 °C and 0–15 min for pressure, temperature and holding time, respectively. The inactivation kinetics was demonstrated with pressure pulse effect followed by static pressure-hold period. A synergistic effect of pressure and temperature was noted on inactivation rate of PPO. Enzyme inactivation kinetics followed a 0.97 order reaction within the range of experimental domain. The activation energy (Ea) and the activation volume (Va) values computed to describe the temperature and pressure dependence of inactivation rate constant (k, min−1), decreased with increase in pressure and temperature, respectively. An empirical model was proposed taking into account the pressure and temperature dependence of Ea and Va, to describe the inactivation rate of PPO as a function of pressure and temperature. A good correlation was established between experimental and predicted values of k.

The Gauss-Eyring model: A new thermodynamic model for biochemical and microbial inactivation kinetics

A new primary model has been developed, using Gaussian distributed populations and Eyrings rate constant for the transition state, to describe inactivation kinetics of enzymes and micro-organisms subjected to heat and chemical treatment. The inactivation of both enzymes and micro-organisms could be associated with the irreversible transition to an inactivated state, as suggested by the Lumry-Eyring model for protein denaturation and enzyme inactivation. The characteristic inactivation model parameters, standard activation enthalpy and entropy, are directly related to the reference temperature and Z-value commonly used for kinetic analysis in food microbiology. An essential feature of the kinetic model is that its parameters, and hence the transition temperature, are treated as stochastic variables. The characteristic line shape of the primary model is the log-normal distribution. The performance of the model was validated, using literature data for enzyme and microbial inactivation over a wide range of temperature and pH.

Thermal inactivation kinetics of β-galactosidase during bread baking.

In this study, β-galactosidase was utilized as a model enzyme to investigate the mechanism of enzyme inactivation during bread baking. Thermal inactivation of β-galactosidase was investigated in a wheat flour/water system at varying temperature-moisture content combinations, and in bread during baking at 175 or 205°C. In the wheat flour/water system, the thermostability of β-galactosidase increased with decreased moisture content, and a kinetic model was accurately fitted to the corresponding inactivation data (R2=0.99). Interestingly, the residual enzyme activity in the bread crust (about 30%) was hundredfold higher than that in the crumb (about 0.3%) after baking, despite the higher temperature in the crust throughout baking. This result suggested that the reduced moisture content in the crust increased the thermostability of the enzyme. Subsequently, the kinetic model reasonably predicted the enzyme inactivation in the crumb using the same parameters derived from the wheat flour/water system. However, the model predicted a lower residual enzyme activity in the crust compared with the experimental result, which indicated that the structure of the crust may influence the enzyme inactivation mechanism during baking. The results reported can provide a quantitative understanding of the thermal inactivation kinetics of enzyme during baking, which is essential to better retain enzymatic activity in bakery products supplemented with heat-sensitive enzymes.

Quick identification of a simple enzyme deactivation model for an extended-Michaelis-Menten reaction type. Exemplification for the D-glucose oxidation with a complex enzyme deactivation kinetics

Abstract One essential engineering problem when developing an industrial enzymatic process concerns the model-based design and optimal operation of the enzymatic reactor based on the process and enzyme inactivation kinetics. For a complex enzymatic system, the “default” used first-order enzyme deactivation model has been proved to lead to inadequate process design or sub-optimal operating policies. The present study investigates if a complex enzyme deactivation can be approximated with simple 1st, 2nd, or a novel proposed model with variable deactivation constant. The approached complex enzymatic system is those of the oxidation of D-glucose to 2-keto-D-glucose in the presence of pyranose 2-oxidase. The necessary “simulated experimental data” have been generated by means of an extended kinetic model from literature used to simulate a batch reactor under well-defined nominal conditions. The proposed enzyme deactivation model has been found to be the best lumping alternative, presenting several advantages: simplicity, flexibility, and a very good adequacy.

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, Un-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.

Impact of pH and Total Soluble Solids on Enzyme Inactivation Kinetics during High Pressure Processing of Mango (Mangifera indica) Pulp.

This study was undertaken with an aim to enhance the enzyme inactivation during high pressure processing (HPP) with pH and total soluble solids (TSS) as additional hurdles. Impact of mango pulp pH (3.5, 4.0, 4.5) and TSS (15, 20, 25 °Brix) variations on the inactivation of pectin methylesterase (PME), polyphenol oxidase (PPO), and peroxidase (POD) enzymes were studied during HPP at 400 to 600 MPa pressure (P), 40 to 70 °C temperature (T), and 6- to 20-min pressure-hold time (t). The enzyme inactivation (%) was modeled using second order polynomial equations with a good fit that revealed that all the enzymes were significantly affected by HPP. Response surface and contour models predicted the kinetic behavior of mango pulp enzymes adequately as indicated by the small error between predicted and experimental data. The predicted kinetics indicated that for a fixed P and T, higher pulse pressure effect and increased isobaric inactivation rates were possible at lower levels of pH and TSS. In contrast, at a fixed pH or TSS level, an increase in P or T led to enhanced inactivation rates, irrespective of the type of enzyme. PPO and POD were found to have similar barosensitivity, whereas PME was found to be most resistant to HPP. Furthermore, simultaneous variation in pH and TSS levels of mango pulp resulted in higher enzyme inactivation at lower pH and TSS during HPP, where the effect of pH was found to be predominant than TSS within the experimental domain. Exploration of additional hurdles such as pH, TSS, and temperature for enzyme inactivation during high pressure processing of fruits is useful from industrial point of view, as these parameters play key role in preservation process design.

Thermal versus Microwave Inactivation Kinetics of Lipase and Lipoxygenase from Wheat Germ

AbstractThe inactivation kinetics of enzymes is an important way to reveal the mechanism of enzyme inactivation by microwaves. In this paper, lipase (LA) and lipoxygenase (LOX) were treated by both microwave irradiation and conventional heating. A temperature‐controlled water bath was used for conventional heating, and a microwave monomode reactor with a simultaneous cooling and precise temperature control system was used for microwave irradiation. The results showed that the effects of both microwave irradiation and conventional heating on enzyme inactivation were enhanced as system temperature increased; LOX was more sensitive to heat than LA. Therefore, compared with conventional heating, microwave irradiation shows no significant differences in inactivating LA or LOX in aqueous solution, and the thermal effect of microwave irradiation is the main cause of enzyme inactivation.Practical ApplicationsWheat germ has a short shelf life because of the high activity of endogenous enzymes. Two relevant endogenous enzymes are involved in the degradation of nutrients, lipase (LA) and lipoxygenase (LOX); therefore, the inactivation of LA and LOX is an essential prerequisite for long‐term storage of wheat germ. Microwave irradiation treatment (MIT) has long been applied to the inactivation of enzymes and has several advantages such as having a minimal effect on food product quality and providing rapid and uniform heating. However, the mechanism of enzyme inactivation by microwaves is still controversial. In this study, wheat germ endogenous enzymes were treated by MIT and conventional heat treatment. Based on residual enzyme activities and first‐order rate of inactivation kinetics, the mechanism of microwave inactivation was demonstrated, offering a theoretical basis for the future application of this technique in grain stabilization.