超高压处理对鲜橙汁中果胶酶及过氧化物酶活性的影响

为推进国内非冷冻浓缩橙汁加工业的发展,对新型超高压杀菌技术对橙汁中酶钝化的效果进行研究,采用200～600MPa超高静压处理鲜榨橙汁,使用紫外分光光度法、滴定法分别测定鲜橙汁中过氧化物酶(POD)和果胶酶(PME)的活性,进行两种酶在常温下超高压钝化酶一级动力学拟合研究。结果表明：在常温条件下200MPa处理10min使两种酶轻微激活,在300～600MPa条件下,随压力和处理时间的增大,两种酶钝化反应明显,且符合一级动力学模型,且果胶酶对压力钝化更加敏感。     

Inactivation Kinetics of Pectin Methylesterase,Polyphenol Oxidase,and Peroxidase in Cloudy Apple Juice under Microwave and Conventional Heating to Evaluate Non-Thermal Microwave Effects

Continuous-flow microwave pasteurization provides important advantages over conventional heat exchangers such as fast volumetric heating, lower tube surface temperature, and possible non-thermal effects that enhance enzymatic and bacterial inactivation. Conventional and microwave-assisted inactivation of pectin methylesterase (PME), polyphenol oxidase (PPO), and peroxidase (POD) in cloudy apple juice were investigated to evaluate non-thermal effects. Experiments were conducted to provide uniform heating with accurate temperature acquisition and similar temperature profiles for conventional and microwave treatments. A two-fraction first-order kinetic model was successfully fitted to the data in a procedure that took into account the whole time-temperature profile instead of assuming isothermal conditions. Predicted inactivation curves for pasteurization at 70 and 80 °C of the cloudy apple juice showed that PME has the highest thermal resistance (residual activity of 30% after 250 s at 80 °C) and that there was no evidence of non-thermal microwave effects on the inactivation of these enzymes.     

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.     

Effect of thermosonication on peroxidase,pectin methylesterase activities and on bioactive compounds in custard apple juice

Custard apple juice faces long term storage challenges such as pink discoloration and bitterness due to peroxidase activity. Therefore, inactivation of peroxidase becomes a necessity for longer shelf life of custard apple and its application in food preparations. Heat processing has certain pitfalls such as loss in nutritional contents of custard apple juice, thereby adversely affecting the sensory quality. In the present study, thermosonication of custard apple juice at a constant temperature of 30?±?1?°C (20 kHz, 67.84 W/cm²) from 0 to 40 min has been studied with respect to peroxidase and pectin methylesterase inactivation. Effect of thermosonication has also been studied on bioactive components such as total phenolics and flavonoids along with enzyme inactivation. Complete inactivation of peroxidase and pectin methylesterase with 0% residual activity was achieved within 40 min and 50 s, respectively. Total phenolics were found to increase from 70.9?±?1.6 to 81.7?±?2.5 mg GAE/g. Sonication was found to have no significant effect (P?>?0.05) on pH, °Brix, titratable acidity. Ascorbic acid content remained relatively unaffected during sonication. Hence, thermosonication may be employed as food processing technique where retention of bioactive components have been achieved along with safety and quality attributes.     

Kinetics of Tomato Pectin Methylesterase Inactivation by Temperature and High Pressure

ABSTRACT: In this study we investigated the inactivation of endogenous pectin methylesterase (PME) in tomato juice during combined high-hydrostatic pressure (ambient to 800 MPa) and moderate temperature (60 to 75 °C) treatments under isobaric and isothermal processing conditions. PME inactivation rates increased with increasing processing temperature, with the highest rate obtained during processing at 75 °C and ambient pressure. Inactivation rates were dramatically reduced as soon as processing pressure was raised. High inactivation rates were again attained when processing pressure exceeded a value of about 700 MPa. Such a behavior was described by considering two parallel mechanisms of inactivation, each one following first order kinetics with its own kinetic parameters.     

Combined osmodehydration and high pressure processing on the enzyme stability and antioxidant capacity of a grapefruit jam

A combined osmodehydration process and high pressure treatment (OD–HHP) was developed for grapefruit jam preservation. The inactivation kinetics of pectin methylesterase (PME) and peroxidase (POD) in the osmodehydrated (OD) jam treated by combined thermal (45–75 °C) and high pressure (550–700 MPa) processes were fitted using special cases of first-order kinetics, the fractional conversion and biphasic models and the Weibull distribution function. No complete inactivation was achieved by any combination of temperature and pressure, and 27–40% and 51–70% of PME and POD, respectively, were pressure-stable fractions. Two PME fractions with different pressure stabilities were observed and kinetic models successfully explained that behavior. POD was found to be very baroresistant and only the labile fraction could be inactivated. The extent of enzyme inactivation was lower in the OD jam in comparison with other food matrices showing a protective effect against the high pressure treatment. The antioxidant capacity was not affected by any treatment. The proposed high pressure preservation processing was able to improve the enzymatic stability of jam obtained by osmotic dehydration without affecting the bioactive content.     

Isolation and Characterization of Pectin Methylesterase from Apple Fruit

Two forms of the enzyme pectin methylesterase are evidenced in the apple (Malus communis). They differ both in their charge and molecular weight. The two enzymes were separated by DEAE-cellulose chromatography. Their molecular weights, determined by gel-filtration, were 55000 and 28000 daltons. The heavier form has been purified at homogeneity and subjected to investigations regarding its activity as a function of the pH and temperature, and determination of its kinetic parameters. The enzyme has a Km value of 1.05 mg/mL for citrus pectin and an optimum activity in the pH range between 6.5-7.5. The enzyme was stable up to 40°C. Incubation for 1 min at 90°C leads to its complete inactivation.     

Yield and quality effects of electroplasmolysis and microwave applications on carrot juice production and storage

Electroplasmolysis as a pretreatment and microwave heating as an alternative to traditional heating were used in carrot juice production and the effects of these electrical methods on juice quality were investigated. Electroplasmolysis and microwave applications were optimized by response surface methodology. Optimum conditions were these: voltage gradient of 22.2 V/cm and treatment time of 60 s for electroplasmolysis, and flow rate of 90 mL/min with power of 900 W for microwave heating. Production of carrot juice was carried out by using these optimum conditions. After production, carrot juice samples were stored for 4 mo at +4 °C and analyses were performed at monthly intervals. As a result of electroplasmolysis, a 9.7% increase in juice yield was obtained. In addition, 100% pectin methylesterase inactivation was found with the microwave heating application. The results showed that the highest values for quality characteristics, such as antioxidant capacity and total pectin, total phenolic, and total carotenoid contents, were obtained with the combined applications of the electrical methods. In addition, these quality characteristics were protected better in the group of combined applications of the electrical methods (electroplasmolysis + microwave) during storage.     

Quality-Related Enzymes in Plant-Based Products: Effects of Novel Food Processing Technologies Part 2: Pulsed Electric Field Processing

Pulsed electric field (PEF) processing is an effective technique for the preservation of pumpable food products as it inactivates vegetative microbial cells at ambient to moderate temperature without significantly affecting the nutritional and sensorial quality of the product. However, conflicting views are expressed about the effect of PEF on enzymes. In this review, which is part 2 of a series of reviews dealing with the effectiveness of novel food preservation technologies for controlling enzymes, the scientific literature over the last decade on the effect of PEF on plant enzymes is critically reviewed to shed more light on the issue. The existing evidence indicates that PEF can result in substantial inactivation of most enzymes, although a much more intense process is required compared to microbial inactivation. Depending on the processing condition and the origin of the enzyme, up to 97% inactivation of pectin methylesterase, polyphenol oxidase, and peroxidase as well as no inactivation have been reported following PEF treatment. Both electrochemical effects and Ohmic heating appear to contribute to the observed inactivation, although the relative contribution depends on a number of factors including the origin of the enzyme, the design of the PEF treatment chamber, the processing condition, and the composition of the medium.     

The Effect of Electric Field on Important Food-processing Enzymes: Comparison of Inactivation Kinetics under Conventional and Ohmic Heating

ABSTRACT: This work deals with the determination of the inactivation kinetics of several enzymes, most of them used as time-temperature integrators in the food industry. The tested enzymes were polyphenoloxidase, lipoxygenase, pectinase, alkaline phosphatase, and p-galactosidase, and the inactivation assays were performed under conventional and ohmic heating conditions. The thermal history of the samples (conventional and ohmically processed) was made equal to determine if there was an additional inactivation caused by the presence of an electric field, thus eliminating temperature as a variable. All the enzymes followed 1st-order inactivation kinetics for both conventional and ohmic heating treatments. The presence of an electric field does not cause an enhanced inactivation to alkaline phosphatase, pectinase, and β- galactosidase. However, lipoxygenase and polyphenoloxidase kinetics were significantly affected by the electric field, reducing the time needed for inactivation. The results of the present work can be used industrially to determine processing effectiveness when ohmic heating technology is applied.     

Pulsed electric fields versus thermal treatment: equivalent processes to obtain equally acceptable citrus juices

Pulsed electric field treatment has been claimed to produce more acceptable chilled citrus juices than those obtained by conventional thermal treatment. The pectin methylesterase activity and the acceptability of nine juices obtained from Clementine mandarins, Valencia oranges, and Ortanique fruits (hybrid of mandarin and orange), untreated, pasteurized (85 degrees C for 10 s), and treated by pulsed electric fields (25 kV/cm for 330 micros), were evaluated. The treatments, selected to reach a similar level of pectin methylesterase inactivation, produced juices that did not differ in acceptability from each other for the three varieties and in all cases were less acceptable than the untreated juice.     

Assessment of the thermal treatment of orange juice during continuous microwave and conventional heating

The effect of continuous-flow microwave heat treatment of orange juice was evaluated by using parameters of quality (pectin methylesterase (PME) inactivation and ascorbic acid, free amino acids, carbohydrates and hydroxymethyl furfural (HMF) contents, as well as degree of browning). Results were compared with those obtained using a conventional tubular heat exchanger having the same heating and cooling phases. The continuous microwave process proved to be an effective system for PME inactivation without the inclusion of a holding phase. In general, no differences were found between the two systems used in the majority of the indicators studied, except that a slight decrease was observed in the most reactive amino acids in the case of conventionally heated orange juice. This fact could be due to overheating of the juice at the tubular heat exchanger. © 1998 Society of Chemical Industry     

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.     

重组猕猴桃果胶甲酯酶抑制剂的抑制活性研究

猕猴桃的果胶甲酯酶抑制剂可有效抑制不同种类植物的果胶甲酯酶活性。主要研究在巴斯德毕赤酵母中成功表达的重组猕猴桃果胶甲酯酶抑制剂(被命名为kWPMEI)的抑制活性。通过研究发现,kWPMEI对桔子果胶甲酯酶的抑制活性最强,其次是胡萝卜,最弱的是番茄。同时kWPMEI对上述三种植物果胶甲酯酶的抑制活性受环境温度和pH的影响比较显著。     

Activities and conformation changes of food enzymes induced by cold plasma: A review

Food endogenous enzymes have impacts on color, texture and flavor of foods during food processing or preservation. Cold plasma is a novel non-thermal food processing technology, which has been extensively studied for contamination elimination and shelf life extension of foods. Particularly, much work has been reported about the effects of cold plasma on enzyme activities and alterations about enzymes conformational structures. It is thus necessary to understand the mechanisms of actions and applications of cold plasma technology in the conformation of food endogenous enzymes. This review focuses on the applications of cold plasma for the inactivation of various endogenous enzymes, including peroxidase, polyphenol oxidase, lysozyme, α-chymotrypsin, alkaline phosphatase, and pectin methylesterase. The activations of several enzymes, such as superoxide dismutase, catalase, and lipase, by cold plasma are also discussed. In addition, this review highlights the transformation of conformational structures including primary and spatial structures induced by chemical reactive species during cold plasma treatments, such as reactive oxygen species and reactive nitrogen species, especially, active sites consisting of prosthetic group and specific amino acids are demonstrated. Both extrinsic and intrinsic factors affecting cold plasma treatments are also described. In general, cold plasma exhibits the ability to activate or inactivate enzymes activities with affecting the conformational structures of enzyme. Further studies should be focused on exploration at molecular level for providing more insight on the interaction mechanism. In addition, equipment and process parameters of cold plasma operation for different fresh food products should be optimized for achieving appropriate control on enzyme variation and obtaining maximum efficiency.     

Carrot pectin methylesterase and its inhibitor from kiwi fruit: Study of activity, stability and inhibition

Carrot pectin methylesterase (PME) and its inhibitor (PMEI) from kiwi fruit were successfully purified by affinity chromatography. Enzyme and inhibitor activity and stability and PME–PMEI complex formation, as influenced by intrinsic product factors (pH and NaCl) and extrinsic process factors (temperature and pressure), were studied. The effect of temperature- or pressure-induced denaturation of PME and PMEI on their respective activities was assessed by estimating inactivation kinetic parameters. PME inactivation obeyed first-order kinetics. The enzyme was rather heat-labile but pressure-stable. PMEI inactivation was best described by a model taking into account a processing-stable PMEI intermediate. The behavior of PME and the PME–PMEI complex at elevated temperature or pressure in the presence of pectin was explored by following methanol formation as a function of treatment time. PME catalytic activity was stimulated up to a certain temperature or pressure level before declining. No conclusive evidence was obtained for a temperature-induced dissociation of the PME–PMEI complex, whereas high pressure exposure caused the complex to separate.

Industrial relevance

PME activity control is a major point of interest in the quest of obtaining high quality plant-derived food products. The current study demonstrates that both traditional thermal processing and novel high hydrostatic pressure processing allow stimulation as well as inactivation of PME and, hence, directing the PME-catalyzed pectin hydrolysis. An alternative or additional approach to control endogenous PME activity (e.g. to obtain cloud-stable juices) is through enzyme inhibition using kiwi PMEI. In this context, pH and NaCl boundaries for application were established, the existence of a temperature- and pressure-stable PMEI intermediate was shown and the PME–PMEI complex was proven not to be dissociated at mild temperature and pressure levels. These observations endorse the possibility of inhibiting undesirable PME activity remaining after mild processing.     

Inhibition of tomato pectin methylesterase by partially purified kiwi pectin methylesterase inhibitor protein

The inhibition of tomato pectin methylesterase (PME) by a recently discovered kiwi pectin methylesterase inhibitor (PMEI) is described. PME was consequently purified by CM Sephadex C-50, Concanavalin A-Sepharose 4 B and Mono S chromatography, and PMEI by Q-Sepharose and Sephacryl S-200 chromatography. Inhibition of tomato PME activity under optimal conditions (0.125 m NaCl, pH 7.5) by partially purified kiwi PMEI (MW of 27 kD, pI ≥ 3.67) was independent of the PMEI/PME ratio between 36 and 61% of the maximal uninhibited activity. the non-competitive inhibition observed was optimal in the pH range of 5 to 7. PMEI was inactivated by heating to 120°C, and showed actinidin-like activity towards N-α-benzyloxycarbonyl-L-lysine p -nitrophenyl ester (CBZ-lys-ONp) and azocasein which was partially inhibited by the protease inhibitor leupeptin.     

Combined high-pressure and thermal treatments for processing of tomato puree: evaluation of microbial inactivation and quality parameters

The effects of combined high-pressure thermal treatments on consistency, viscosity, colour, lycopene content, enzyme activity and micro-organisms were determined, and compared to conventional pasteurisation and sterilisation processes of tomato puree. High-pressure processing at ambient temperature (HPP) improved the colour and viscosity compared to heat pasteurisation, while the water binding capacity and lycopene content were unaffected by HPP. Products treated at 700 MPa, 20 °C resulted in inactivation of the natural flora to a level below the detection limit. After pressure treatment and during chilled storage a increase in viscosity was observed. HPP caused partial inactivation of polygalacturonase (70%), but activation of pectin methylesterase. After high-pressure sterilisation treatments combined with elevated starting temperatures (≥80 °C, HPS, one or two pulses) an ambient stable product was obtained. HPS (one pulse, 700 MPa, 30 s, 90 °C) reduced B. stearothermophilus spore contamination level in inoculated meatballs in tomato puree with at least 4.5 log units. HPS resulted in more than 99% inactivation of polygalacturonase and pectin methylesterase. HPS resulted in a lower viscosity compared to conventional sterilised samples, whereas the water binding capacity was improved. Colour appreciation was improved and lycopene content was retained compared to a 40% loss after conventional sterilisation.     

High Pressure and Temperature Effects on Enzyme Inactivation in Strawberry and Orange Products

High hydrostatic pressure treatment (50-400 MPa) combined with heat treatment (20–60°C) effects on peroxidase (POD), polyphenoloxidase (PPO) and pectin methylesterase (PME) activities of fruit-derived products were studied. Assays were carried out on fresh orange juice and strawberry puree. Pressurization/depressurization treatments caused a significant loss of strawberry PPO (60%) up to 250 MPa and POD activity (25%) up to 230 MPa, while some activation was observed for treatments carried out in 250–400 MPa range for both enzymes. Optimal inactivation of POD was using 230 Mpa and 43°C in strawberry puree. Combinations of high pressure and temperature effectively reduced POD activity in orange juice (50%) to 35°C. The effects of high pressure and temperature on PME activity in orange juice were very similar to those for POD.     

A review of pectin methylesterase inactivation in citrus juice during pasteurization

BackgroundPectin, naturally found in citrus, plays a key role in the quality of the obtained juices. Pectin methylesterase enzyme (PME) influences the cloud stability, viscosity, color, mouth feeling and flavor of the juices by de-esterification of pectin. Iinactivation of PME is introduced as a pasteurization index in citrus juices, due to its higher thermal resistance than the spoilage microorganisms.Scope and approachInactivation of PME using different thermal (conventional, microwave and ohmic heating) and non-thermal (pulsed electric field, high pressure processing and high pressure carbon dioxide) processes is important in juice production. The aim of this study was to review the effect of these processing methods on the PME inactivation in different citrus juices.Key finding and conclusionUsing non-thermal methods in combination with moderate thermal methods can be more effective in PME inactivation with minimum loss in citrus juice quality.