Changes in microorganism, enzyme, aroma of hami melon (Cucumis melo L.) juice treated with dense phase carbon dioxide and stored at 4 °C

The effects of dense phase carbon dioxide (DP-CO₂) treatment of 8, 15, 22, 30 and 35 MPa for 5 min, 15 min, 30 min, 45 min, 60 min at 35 °C, 45 °C, 55 °C, 65 °C on microorganism, enzyme, and aroma compounds in hami melon juice during storage at 4 °C for 4-weeks were investigated. Meanwhile, the color, browning degree, and Vitamin C were also studied. The DP-CO₂ treatment had significant effects on inactivation of microorganism and enzyme. It was indicated that higher pressure caused more inactivation of microbial total count and enzyme activity. When it reached 35 Mpa, 55 °C, 60 min, the microorganism was totally inactivated. The least residual activity of polyphenol oxidase (PPO), peroxidase (POD), and lipoxygenase (LOX) was 25.26%, 38.46 and 0.02% at 35MP, respectively. The restoration of PPO, POD and LOX residual activity after DP-CO₂ treatment was also observed, which was dependent on the pressure level. The aroma compounds were less affected after being treated with DP-CO₂, and the flavor of the melon juice was close to the fresh juice after storage at 4 °C for 4 weeks and did not produced cook off-odor. The changes of lightness L and browning degree A during storage were well fitted to a first-order kinetic model. The Vitamin C concentration decreased by DP-CO₂ processing, but this loss was lower than of the untreated sample.

Industrial relevance

Hami melon is highly appreciated for its nutritional quality and special flavor. The flesh of melon is heat sensitive, the sensitive nutrients, color and aromatic profile will be spoiled greatly or off-odour when it was produced with high temperature treatment. Dense phase carbon dioxide processing (DP-CO₂) is important to find an innovative food process to inactivate the enzyme and microorganism and protect the nutrient and unique flavor. In this study, the data proved that DP-CO₂ processing is a promising non-thermal alternative pasteurization to preserved fresh-squeezed melon juice.     

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。     

超高压处理对哈密瓜汁品质酶和微生物的影响

本文对不同超高压处理后的哈密瓜汁中的POD、PPO和LOX的活性进行了测定，同时对哈密瓜汁中微生物及其对象芽孢菌和安全性评价菌E．coli的耐压性进行了实验。研究结果表明：哈密瓜汁经500MPa、20min处理后，其POD、PPO和LOX的残留活力分别为81％、8．06％和6．84％，在同等处理条件下，对象芽孢菌耐压，大肠杆菌不耐压且降低了5个对数级，符合美国FDA鲜榨果蔬汁非热力杀菌的安全要求，哈密瓜汁中的细菌总数≤100CFU／ml，符合我国饮料的卫生标准。     

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.     

Steam blanching effect on polyphenoloxidase,peroxidase and colour of mango (Mangifera indica L.) slices

The heat stability of peroxidase (POD) and polyphenoloxidase (PPO) was investigated in mango (Mangifera indica L.) slices, and the relative colour was studied after different steam blanching times. There was complete inactivation after 5 min for POD and 7 min for PPO. Steam blanching of 3 min gave residual activity of 2.85% and 8.33% for PPO and POD, respectively, and when compared with samples blanched for 5 min had no effect on colour over 20 days of storage. Correlation was found between activities of PPO, POD and colour change over 20 days. After 7 min steam blanching the browning index was stable but less than at 3 and 5 min because non-enzymic browning had occurred. This research suggests that yellowness (b) and lightness (L) values contribute positively to the browning index (BI), compared to redness (a).     

Use of response surface methodology to study the combined effects of UV-C and thermal processing on vegetable oxidative enzymes

The combined thermal (25–65 °C) and ultraviolet processing (UV-C) effects on lipoxygenase (LOX), peroxidase (POD) and polyphenoloxidase (PPO) at different pH values (4.0–7.0) were studied using a central composite design. An initial screening design revealed that all factors had a significant effect on enzymatic activity except wavelength which showed a negligible effect. A synergistic effect was found between temperature and UV exposure time for POD and PPO and between pH and exposure time for LOX. LOX enzyme was affected by acidic conditions. POD was UV-C labile whereas PPO was the most UV-C resistant enzyme but was thermolabile. Second-order polynomial equations indicated that enzyme activities were inactivated after exposure to 58.2 mJ/cm² UV at 60 °C or higher temperatures at any pH condition. Combination of UV and thermal processing allowed the use of low energy/doses to obtain complete enzymatic inactivation. This study may serve as a basis to design UV-C processes for the inactivation of enzymes in liquid matrices.     

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.     

Kinetic characterisation and thermal inactivation study of polyphenol oxidase and peroxidase from table grape (Crimson Seedless)

Polyphenol oxidase (PPO) and peroxidase (POD) were extracted from a table grape (Crimson Seedless) using Triton X-114 and characterized using spectrophotometric methods. Both PPO and POD were activated by acid shock. However, in the presence of the anionic detergent sodium dodecil sulphate (SDS), PPO was activated whereas POD was inactivated. The enzymes were kinetically characterized and both followed Michaelis–Menten kinetics, although with different values of their kinetic parameters. The V_m/K_m ratio showed that Crimson Seedless grape PPO presents a similar affinity for 4-tert-butyl-catechol (TBC) whether activated by acid shock (0.018 min⁻¹) or SDS (0.023 min⁻¹). With regards to POD, the K_m and V_m values for 2,2′-azinobis(3-ethylbenzothiazolinesulphonic acid) (ABTS) were 0.79 mM and 1.20 μM/min, respectively. In the case of H₂O₂, the K_m and V_m value were 0.4 mM and 0.93 μM/min, respectively. PPO and POD showed similar thermostability, losing >90% of relative activity after only 5 min of incubation at 78 °C and 75 °C, respectively. In addition, PPO´s activation energy was similar to that obtained for POD (295.5 kJ/mol and 271.9 kJ/mol, respectively).     

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.     

Kinetic Parameters for the Thermal Inactivation of Peroxidase and Lipoxygenase in Precooked Frozen Brassica Species

Thermal inactivation of peroxidase (POD) and lipoxygenase (LOX), both enzymes present in broccoli and Brussels sprouts, is required before freezing, to obtain high‐quality precooked frozen vegetables. Rate constants of a 1st‐order biphasic model for the heat‐labile and heat‐resistant POD and LOX isoenzymes were determined at different temperatures (75, 80, and 90 °C) and the corresponding activation energies were estimated using nonlinear regressions. In the case of Brussels sprouts, the activation energies for the resistant and labile fractions were 56.3 and 62.5 kJ/mol for POD and 63.7 and 65.8 kJ/mol for LOX, respectively. For Brussels sprouts, different precooking times were tested to analyze the effect of residual enzyme activity on quality parameters and sensory attributes, after a frozen storage of 4 mo at ?20 °C. A significant reactivation of enzyme activity after frozen storage was observed (especially in the case of POD) for short precooking times (<6 min) leading to low‐quality parameters at the interior zone of the vegetable. A precooking time of 6 min at 90 °C allowed an adequate inactivation of LOX and POD obtaining a high‐quality final frozen vegetable. A sensory analysis confirmed the global acceptability of the product. The obtained results are relevant to define the precooking stage conditions in the production of frozen cruciferous vegetables.     

Highly effective inactivation of anti-nutritional factors (lipoxygenase,urease and trypsin inhibitor) in soybean by radio frequency treatment

Radio frequency (RF) treatment at 27.12 MHz was employed to inactivate the anti-nutritional factors (ANFs), including lipoxygenase (LOX), urease and trypsin inhibitor to minimise deleterious effects caused by conventional hot-air heating. The effects of RF heating on the ANF activities, physicochemical properties and processing quality of intact soybean were investigated. The results revealed that ANF activities were effectively inactivated within 300 s by RF heating and inactivation rates of LOX, urease and trypsin inhibitor were 95.2% (270 s), 93.4% (285 s) and 89.4% (300 s), respectively. In addition, RF heating improved physicochemical properties and processing quality of soybean products when the trypsin inhibitor was inactivated. On the contrast, conventional thermal treatment significantly decreased functional properties and led to the formation of greater aggregates. Our results may provide a novel method to inactivate endogenous enzyme in crops without negative impact on processing quality.     

Thermosonication: a potential technique that influences the quality of grapefruit juice

Thermosonication (TS) is an emerging nonthermal processing technique used for the liquid food preservation and is employed to improve the quality and acceptability of grapefruit juice. In this study, fresh grapefruit juice samples were subjected to TS treatment in an ultrasonic cleaner with different processing variables, including temperature (20, 30, 40, 50 and 60 °C), frequency (28 kHz), power (70%, 420 W) and processing time (30 and 60 min) for bioactive compounds, inactivation of enzymes pectin methylesterase (PME), peroxidase (POD) and polyphenolase (PPO) and micro‐organisms (total plate count, yeasts and moulds). The micro‐organism activity was completely inactivated in the treatment (60 °C for 60 min). The TS treatment at 60 °C for 60 min exposure reduced PME, PPO and POD activity by 91%, 90% and 89%, respectively. Results indicate that the advantages of TS for grapefruit juice processing at low temperature could enhance the inactivation of enzymes and micro‐organisms and it can be used as a potential technique to obtain better results as compared to alone .     

不同氧分压贮藏条件下金针菇衰老相关酶活性的研究

研究不同氧分压条件下金针菇贮藏过程中抗氧化相关酶活性的变化及其对保鲜效果的影响。在聚乙烯塑料袋中放入金针菇100g、膨润土吸湿剂8g、活性炭6g,分别按0、20%、50%、70%、80%、90%、100%氧气分压充气包装,于1～3℃冷藏,每隔5～7d取样检测。结果显示：80%氧气分压试验组的保鲜效果最好;在贮藏的最初2周,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、脂氧合酶(LOX)等与抗氧化、清除自由基相关的酶活性明显升高,多酚氧化酶(PPO)、过氧化物酶(POD)、纤维素酶(CMCase)等酶活性降低;贮藏2周后SOD、CAT、LOX等酶活性明显减弱,POD、PPO等酶活力明显增强。表明SOD、CAT、LOX等抗氧化酶活性的减弱,导致菇体内O2－ ·、 ·OH、过氧化物等逐渐积累,细胞膜结构受损,细胞内容物外渗增多;纤维素酶对细胞壁降解,导致组织软化出水,酶促褐变活力增强,是致使金针菇衰老、加速变质、品质降低的重要原因。     

Effect of nitric oxide on pericarp browning of harvested longan fruit in relation to phenolic metabolism

The effects of nitric oxide (NO) on enzymatic browning of harvested longan fruit in relation to phenolic metabolisms were investigated. Fruits were dipped for 5 min in 1 mM sodium nitroprusside (SNP), a nitric oxide donor, then packed in 0.03 mm thick polyethylene bags, and finally stored for 6 days at 28 °C. Changes in pericarp browning and pulp breakdown were evaluated, while total phenol content, activities of phenolic-associated enzymes, polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), and concentrations of total soluble solids, titratable acidity and ascorbic acid were measured. SNP treatment delayed pericarp browning, inhibited activities of PPO, POD and PAL and maintained a high total phenol content of longan fruit during storage. Furthermore, NO showed a significant inhibition of the in vitro activities of PPO and POD, indicating that the beneficial effect of NO was direct. Moreover, application of NO resulted in a lower pulp breakdown and maintained relatively high levels of total soluble solids and ascorbic acid.     

Inactivation of peroxidase and lipoxygenase in carrots, green beans, and green peas by combination of high hydrostatic pressure and mild heat treatment

The efficiency of high hydrostatic pressure (HHP) with the combination of mild heat treatment on peroxidase (POD) and lipoxygenase (LOX) inactivation in carrots, green beans, and green peas was investigated. In the first part of the study, the samples were pressurized under 250–450 MPa at 20–50 °C for 15–60 min. In the second part, two steps treatments were performed as water blanching at 40–70 °C for 15 and 30 min after pressurization at 250 MPa and 20 °C for 15–60 min. Carrot POD was decreased to 16% residual activity within the first 30 min at a treatment condition of 350 MPa and 20 °C and then it decreased to 9% at 60 min. When the carrots were water blanched at 50 °C for 30 min after HHP treatment of 250 MPa at 20 °C for 15 min, 13% residual POD activity was obtained. For green beans, the most effective results were obtained by two steps treatment and approximately 25% residual POD activity was obtained by water blanching at 50 °C for 15 min after pressurization at 250 MPa and 20 °C for 60 min. An effective inactivation of POD in green peas was not obtained. For carrots, LOX activity could not be measured due to very low LOX activity or the presence of strong antioxidants such as carotenoids. After pressurization at 250 MPa and 20 °C for 15 or 30 min, water blanching at 60 °C for 30 min provided 2–3% residual LOX activity in green beans. The treatment of 250 MPa for 30 min and then water blanching at 50 °C for 30 min provided 70% LOX inactivation in green peas.     

Exogenous salicylic acid inhibits browning of fresh-cut Chinese water chestnut

The potential usage of salicylic acid (SA) as a powerful anti-browning agent in fresh-cut Chinese water chestnut (CWC) was investigated. The fresh-cut CWC were dipped for 1 min in solutions of 0, 1, 2 or 4 mM SA, then placed in trays over-wrapped with plastic films, and finally stored at 4 °C. Changes in color, eating quality, and disease incidence were evaluated, while activities of phenol-associated enzymes, polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase (PAL), and concentrations of total soluble solid, titratable acidity and ascorbic acid were measured. SA treatment delayed discoloration, maintained eating quality with higher content of the quality attributers, and reduced activities of or delayed the increases in activities of PPO, POD and PAL in fresh-cut CWC. However, SA had no significant inhibition of the activities of PPO and POD in an in vitro test, indicating that the beneficial effect of SA was indirect. Further research is needed to elucidate the inhibition of the surface browning of the fresh-cut CWC by SA.     

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.     

The role of polyphenol oxidase and peroxidase in the browning of water caltrop pericarp during heat treatment

The mechanism of browning involving enzymatic browning was investigated in the pericarp of water caltrop, an Asian vegetable popular for its taste and medicinal properties. Polyphenol oxidase (PPO) and peroxidase (POD) activities were determined in pericarp at various times and temperatures. Water caltrop consisted of 44.22% moisture content, 37.23% crude fibre, and 2.63% crude protein. PPO and POD activities dropped from 62 and 38 units/g sample, respectively, as water temperature was increased from 30 to 80 °C. Optimum pH and temperature for PPO activity was at pH 5.0, 25–45 °C, and POD activity peaked at 60 °C. High PPO and POD activities at 40–50 °C resulted in degradation of phenolic compounds, which led to increased aggregation of browning pigments and discolouration (lower L-values) of the pericarp. Enzymatic browning was determined as the major factor in the browning discolouration of heat-treated water caltrop pericarp.     

Color,texture and enzyme activities of Hypsizygus marmoreus as affected by heating combined with color protection and hardening

Different pretreatments can affect the quality of Hypsizygus marmoreus (HM). In this study, the effects of thermal treatment or combined with color protection and hardening on color, texture, microstructure, polyphenol oxidase (PPO) and Peroxidase (POD) C-1: Explain the first time? activities, and the volatile components of HM were investigated. The results showed compared with the control group, both heating group (H group) and color protection, hardening and heating (CHH group) significantly enhanced the color difference, and reduced all texture parameters as well as PPO and POD activities. Additionally, in two treatment groups, the structure of HM became loose, the amounts of alcohols, ketones, and aldehydes were changed significantly. Compared with the H group, the color difference and PPO activities in CHH group were significantly decreased, while all texture parameters were increased significantly, the fibers of HM showed regular arrangement. In conclusion, the combination of CHH performed better effects on hardness, chewiness, microstructure, and enzymes activities related to enzymatic browning.     

High pressure processing of cocoyam,Peruvian carrot and sweet potato: Effect on oxidative enzymes and impact in the tuber color

High pressure processing (HPP) is a non-thermal technology used to activate or inactivate enzymes. This study investigated the effects of HPP (600 MPa for 5 or 30 min at 25 °C) on cocoyam, Peruvian carrot and sweet potato color, and the polyphenoloxidase (PPO) and peroxidase (POD) activities in tuber cubes, puree, and enzyme extract subjected to HPP. The results showed enzyme inactivation by HPP in cocoyam (up to 55% PPO inactivation in puree and 81% POD inactivation in extract) and Peruvian carrot (up to 100% PPO and 57% POD inactivation the extract). In contrast, enzyme activation was observed in sweet potato (up to 368% PPO and 27% POD activation in puree). The color results were compatible to enzyme activity: the color parameters remained unchanged in cocoyam and Peruvian carrot, which showed high PPO and POD inactivation after HPP. Furthermore, the impact of HPP on the enzymes was influenced by the matrix in which HPP was carried out, evidencing that the enzyme structure can be protected in the presence of other food constituents.Industrial relevanceThe enzymes PPO and POD are an important concern for vegetable processing, due its ability to induce browning after vegetables are cut. The HPP at 600 MPa for 5 or 30 min can be used to inactivate these enzymes in cocoyam and Peruvian carrot, guaranteeing the color and freshness of the tubers similar to the fresh cut vegetable.