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.     

The influence of the malaxation temperature on the activity of polyphenoloxidase and peroxidase and on the phenolic composition of virgin olive oil

The effect of the malaxation temperature under sealed conditions on the qualitative and quantitative composition of the phenolic compounds in virgin olive oils produced from four Italian cultivars was assessed for two atmospheric conditions. In both cases, the results show a positive relationship between temperature and the concentration of the derivatives of the secoiridoid aglycones; the effect of the temperature on the oxidoreductases that promote oxidation (polyphenoloxidase and peroxidase) was investigated to determine their optimal temperatures and thermal stability. While olive peroxidase (POD) showed the highest activity at 37 °C and high stability in the temperature range tested, polyphenoloxidase (PPO) exhibited the optimum activity at approximately 50 °C, but showed low stability at 40 °C, with a large variation in stability according to the olive cultivar. These results may contribute to an understanding of the increase in the phenol concentration found in virgin olive oils obtained following higher temperatures of malaxation.     

Peroxidase and Polyphenoloxidase Activities in Papaya During Postharvest Ripening and After Freezing/Thawing

Changes in peroxidase (POD) and polyphenoloxidase (PPO) activities of papaya (Caricu papava), cv Sunrise, during ripening, freezing and short frozen storage were studied. Fruits were stored at 14°C and 85 90 % RH until maturity for processing was reached (about 21 days). Fruit were frozen cryogenically and frozen slices were stored at — 18°C. POD activity increased in pulp tissue up to the ripe stage, showing a maximum value after 7 days cold storage. Similarly, PPO activity showed an important increase (4 × initial value) on the same date. The quantity of extractable proteins was at a maximum after 15 days storage at 14°C. Freezing and frozen storage (-18°C) produced an increase of POD activity while EPO activity was only slightly affected.     

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).     

Broccoli (<Emphasis Type="Italic"> Brasica oleracea</Emphasis>) treated under pressure-shift freezing process

The experiment was carried out on broccoli to study the freezing effects under pressure using the pressure shift freezing treatment (PSF). The treatment was applied to samples that were in polyethylene bags, sealed under a vacuum at 180 MPa, –16°C and 210 MPa, –20°C and in samples blanched for 2 and 3 min. Protein content, peroxidase (POD), polyphenoloxidase (PPO) activity and structure were analysed. The final results showed that the proteins decreased after the treatments. Five heat-cold-pressure shock proteins of 50, 43, 39, 36 and 20 KDa were found. The assayed conditions were not enough to inactivate the POD and PPO enzymes. Four POD-isozymes of 110, 80, 50 and 39 KDa were detected and the highest activity was for the 80 KDa isoform. The sensorial test revealed that after 30 days of frozen storage at –20 °C the flavour of broccoli was not acceptable to consumers, but the texture remained quite firm. Cryo-Sem revealed that the vacuole membrane was destroyed and an internal disorganised cell was observed after the PSF treatment.     

Limiting the deterioration of mango fruit during storage at room temperature by oxalate treatment

Effects of oxalate on the incidence of decay and ripening in mango fruit, and its physiological effects on the peel and flesh of mango were investigated after mango fruit (Mangifera indica L.) were dipped in different oxalate solutions for 10 min and then stored at 25 °C. Oxalate application decreased the incidence of decay and delayed the ripening process in mango fruit during storage. Potassium oxalate treatment resulted in increased activities of peroxidase (POD) in both the peel and the flesh and polyphenol oxidase (PPO) in the peel, without activation of phenylalanine ammonia-lyase activity, and elevated total phenolic content in the peel. The physiological effects of oxalate in increasing activities of POD and PPO and elevating total phenolic level could be involved in induced resistance of mango fruit against postharvest disease. Oxalate application could be a promising method to suppress postharvest deterioration and extend the useful shelf-life of mangoes.     

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.     

甜玉米速冻前热烫工艺研究

对甜玉米速冻前热烫工艺进行了研究,通过PPO、POD等酶活性、可溶性糖和Vc等指标的检测分析和感官评价,探讨了水煮热烫处理、蒸气热烫处理和微波热烫处理等热烫处理方式、热烫处理时间等因素对速冻甜玉米品质的影响。试验结果表明,适宜的热烫处理工艺(蒸气热烫20min)能减少可溶性糖和Vc的损失,同时钝化PPO和POD酶活性,提高速冻甜玉米的品质。     

Enzymes associated with blackheart development in pineapple fruit

The involvement of browning enzymes, polyphenoloxidase (PPO), peroxidase (POD) and phenylalanine ammonia-lyase (PAL) in blackheart development was investigated in pineapple fruit (Ananas comosus, Smooth Cayenne) following low temperature storage. An increase in PPO activity was related to the incidence of blackheart symptoms, both temporally and spatially. Fruit maturity significantly affected blackheart susceptibility; immature and over-mature fruits developed less blackheart injury than mature fruit. The effect of maturity on blackheart susceptibility was highly correlated to the response of PPO activity to chilling. POD showed no significant change after chilling. Enhanced PAL activity was observed during chilling at 6, 13 and 18 °C. Chilling (6, 13 and 18 °C) also inhibited the increase of ascorbate peroxidase activity observed in the fruit stored continuously at 25 °C, but had less effect on catalase activity. The results indicate that the development of blackheart symptoms in pineapple fruit results from the disturbance of a number of metabolic processes that occur at sub-ambient temperatures.     

Characterization of polyphenoloxidase (PPO) and total phenolic contents in medlar (Mespilus germanica L.) fruit during ripening and over ripening

Characterization of polyphenoloxidase (PPO) enzyme and determination of total phenolic concentrations during fruit ripening and over ripening in medlar (Mespilus germanica L.) were determined. During ripening, PPO substrate specificity, optimum pH and temperature, optimum enzyme and substrate concentrations were determined. Among the five mono- and di-phenolic substrates examined ((p-hydroxyphenyl) propionic acid, l-3,4-dihydroxyphenylalanine, catechol, 4-methylcatechol and tyrosine), 4-methylcatechol was selected as the best substrate for all ripening stages. A range of pH 3.0–9.0 was also tested and the highest enzyme activity was at pH 7.0 throughout ripening. The optimum temperature for each ripening stage was determined by measuring the enzyme activity at various temperatures over the range of 10–70 °C with 10 °C increments. The optimum temperatures were found to be 30, 20 and 30 °C, respectively, for each ripening stage. Optimum enzyme and substrate concentrations were found to be 0.1 mg/ml and 40 mM, respectively. The V_max and K_m value of the reaction were determined during ripening and found to be 476 U/mg protein and 26 mM at 193 DAFB (days after full bloom) – stage 1, 256 U/mg protein and 12 mM at 207 DAFB – stage 2, 222 U/mg protein and 8 mM at 214 DAFB – stage 3. For all ripening stages sodium metabisulfite markedly inhibited PPO activity. For stage 1 of ripening, Cu²⁺, Hg²⁺ and Al³⁺, for stage 2, Cu²⁺ and Hg²⁺, and for stage 3, Cu²⁺, Hg²⁺, Al³⁺ and Ca²⁺ strongly inhibited diphenolase activity. Accordingly, it can be concluded that as medlar fruit ripen there is no significant changes in the optimum values of polyphenoloxidases, although their kinetic parametres change. As the fruit ripening progressed through ripe to over-ripe, in contrary to polyphenoloxidase activity, there was an apparent gradual decrease in total fruit phenolic concentrations, as determined by using the aqueous solvents and water extractions.     

Combined effect of temperature and pulsed electric fields on apple juice peroxidase and polyphenoloxidase inactivation

Pulsed electric fields (PEF) were applied to freshly prepared apple juice using a laboratory scale continuous PEF system to study the feasibility of inactivating peroxidase (POD) and polyphenoloxidase (PPO). Square wave PEF using different combinations of electric field strength, pre-treatment temperature and treatment time were evaluated in this study and compared to conventional pasteurisation (72 °C; 26 s). Inactivation curves for the enzyme were plotted for each parameter and inactivation kinetics were calculated. Results showed the highest level of decrease in the enzymatic activity of 71% and 68%, for PPO and POD, respectively, were obtained by using a combination of preheating to 50 °C, and a PEF treatment time of 100 μs at 40 kV/cm. This level of inactivation was significantly higher (P < 0.05) than that recorded in juice processed by conventional mild pasteurisation where the activity of PPO and POD decreased by 46% and 48%, respectively. The kinetic data for the inactivation of both enzymes could be described using a 1st-order model (P < 0.001).     

Biocontrol of postharvest gray and blue mold decay of apples with Rhodotorula mucilaginosa and possible mechanisms of action

The efficacy of Rhodotorula mucilaginosa against postharvest gray mold, blue mold and natural decay development of apples and the possible mechanisms involved were investigated. The decay incidence and lesion diameter of gray mold and blue mold of apples treated by R. mucilaginosa were significantly reduced compared with the control fruits, and the higher concentration of R. mucilaginosa, the better the efficacy of the biocontrol. R. mucilaginosa also significantly reduced the natural decay development of apples following storage at 20 °C for 35 days or at 4 °C for 45 days followed by 20 °C for 15 days. Germination and survival of spores of Penicillium expansum and Botrytis cinerea were markedly inhibited by R. mucilaginosa in an in vitro test. Rapid colonization of the yeast in apple wounds was observed whether stored at 20 °C or 4 °C. In apples, the activities of peroxidase (POD) and polyphenoloxidase (PPO) were significantly induced and lipid peroxidation (malondialdehyde (MDA) content) was highly inhibited by R. mucilaginosa treatment compared with those of the control fruits. All these results indicated that R. mucilaginosa has great potential for development of commercial formulations to control postharvest pathogens on fruits. Its modes of action were based on competition for space and nutrients with pathogens, inducement of activities of defense-related enzymes such as POD, PPO and inhibition of lipid peroxidation (MDA content) of apples, so as to enhance the resistance and delay the ripening and senescence of apples.     

Synergistic effect of thermosonication to reduce enzymatic activity in coconut water

In order to minimize the depreciation of coconut water quality caused by the conventional heat treatment, non-conventional preservation methods have been studied. Ultrasound technology can be suitable as an alternative thermal treatment for several food matrices, due to its efficiency in microbiological and enzymatic inactivation. The objective of this work was to evaluate the effect of the thermosonication to reduce the enzymatic activity in coconut water and optimize the optimal operating parameters (amplitude and time) and corresponding specific acoustic energy required for the complete enzymatic inactivation. Ultrasound presented an additive effect to the heat treatment on the inactivation of the polyphenoloxidase (PPO) and peroxidase (POD). It was verified that from 500 to 550 mW/mL a great inactivation of the PPO and POD is achieved, already for a complete inactivation of these enzymes the specific acoustic energy necessary is 655.80 mW/mL. It was also verified that the both enzymes should be taken into account for a US process since PPO and POD presented very similar resistance.Industrial relevanceThis work demonstrates the great potential of the use of thermosonication for the processing of heat-sensitive products, such as coconut water. Thought this innovative processing the food industry will be able to reduce the damages caused by the heat and consequently to obtain higher quality products.     

EFFECT OF HIGH PRESSURE TREATMENTS ON PEROXIDASE AND POLYPHENOLOXIDASE ACTIVITIES

The effects of hydrostatic pressure of 1–9 kbar on peroxidase (POD) and polyphenoloxidase (PPO) activity were studied. Trials were carried out on crude enzymatic extracts obtained from carrots and apples as the sources of POD and PPO, respectively, with different pH values (from 4.5 to 7.0). In both cases pressurization caused a remarkable enzyme activation after 1-min treatments carried out at 3–5 kbar, while a complete enzyme inactivation was observed at 9 kbar. The pH values of the crude enzymatic extracts seemed to affect the degree of activation and/or inactivation due to different pressure levels. A slight recovery in enzyme activity was measured for POD samples during the first 24 h of storage after the treatments.     

Variations of colour,polyphenol oxidase and peroxidase activities during the production of low temperature dried pasta in various durum wheat genotypes

This research was conducted to determine the effect of milling and pasta making on yellow pigment content, soluble brown pigment content, polyphenol oxidase (PPO) activity and peroxidase (POD) activity in semolina and pasta derived from ten durum genotypes. Results showed that pasta colour was impacted more by the milling than by pasta making. The loss of yellow pigment content, POD activity, PPO activity and soluble brown pigment content due to milling was 1.3×, 4.4×, 6.2× and 17.5×, respectively, greater than losses due to pasta making. Dry pasta contained 62%, 31%, 16% and 25% of the original yellow pigment content, brown pigment content and PPO activity and POD activity in the grain, respectively. Semolina yellow pigment content had a positive effect on pasta colour while semolina protein content, ash content and speck count had negative effects. These results indicate the importance of selecting genotypes that have high yellow pigment content and excellent milling qualities.     

Effect of hexanal vapour on longan fruit decay,quality and phenolic metabolism during cold storage

The effects of postharvest treatment with hexanal vapour on longan fruit decay, quality, hexanal residue, phenolic compound content, and polyphenoloxidase (PPO) and peroxidase (POD) activities were studied during storage at 5 °C for 30 days. Hexanal exposure for 2 h at 900 μL L^?1 before cold storage reduced the percentage of fruit with decay and was deemed the optimum treatment. Hexanal exposure resulted in a pericarp that was more reddish brown and less intense in colour. Hexanal residue in the pericarp and aril of fumigated fruit was several fold higher than that of nonfumigated fruit, although levels were low at the end of cold storage. Electrolyte leakage of pericarp increased during 5 °C storage and was further increased by hexanal exposure. Hexanal reduced pericarp phenolic content, and increased PPO and POD activities. Overall, use of hexanal vapour reduced postharvest disease of longan fruit but increased the likelihood of pericarp browning.     

水溶性壳聚糖涂膜处理对采后绿芦笋贮藏品质的影响

研究了0.50%的水溶性壳聚糖(WSC)和羧甲基壳聚糖(CMC)分别对在2℃贮藏条件下采后绿芦笋的涂膜保鲜。通过感官品质、失重率、抗坏血酸含量、叶绿素含量、丙二醛(MDA)含量、总酚含量、多酚氧化酶(PPO)等生理生化指标的测定,以未经处理的绿芦笋为对照。结果表明:经0.50%的CMC和WSC进行涂膜处理,均可有效地降低芦笋的失重率,维持其硬度,延缓其抗坏血酸和叶绿素营养物质的消耗。而且CMC处理可以明显抑制绿芦笋总酚含量的下降及其PPO、POD和SOD活性的变化,其感官品质也明显优于对照组,使绿芦笋的贮藏时间延长至35 d。     

Effect of nanocomposite-based packaging on postharvest quality of ethylene-treated kiwifruit (Actinidia deliciosa) during cold storage

A novel nanocomposite-based packaging (NCP) was prepared by blending polyethylene (PE) with nano-Ag, nano-TiO₂ and montmorillonite. The effects of NCP on the quality parameters of ethylene-treated mature kiwifruit were investigated during the 42 d of storage at 4 °C. The results showed that adding nanoparticles to the PE significantly decreased the oxygen, water vapor permeability and longitudinal strength, and inhibited spore germination. The weight loss, softening, color variation and soluble solid content of kiwifruit were significantly inhibited by 22.67%, 124.84%, 23.46% and 14.42% respectively, which indicated that NCP could delay the ripening of kiwifruit. However, ascorbic acid and total phenols contents in NCP-treated fruit were increased compared with the controls. Additionally, kiwifruit in NCP exhibited 57.44% lower headspace ethylene concentration, 29.44% for malondialdehyde (MDA), lower polyphenol oxidase (PPO) activity and higher peroxidase (POD) activity than the controls. These results suggest that NCP may be a useful technique to reduce fruit decay and maintain quality in kiwifruits during postharvest storage.     

POLYPHENOLOXIDASE ACTIVITY, BROWNING POTENTIAL AND PHENOLIC CONTENT OF PEACHES DURING POSTHARVEST RIPENING

Changes in the polyphenoloxidase (PPO) activity and the phenolic content of peaches (Prunus persica cv. Premier) during postharvest ripening were studied. The fruits were stored at 12 or 25C for up to 15 days. The quantity of extractable proteins was maximum at 6–10 days storage at 25 and 12C, coinciding with the onset of the yellowness in the fruits. The PPO activity increased up to the ripening stage, showing a maximum value at 8 days of storage. This was coincident with the maximum degree of browning as evaluated by the absorbance at 440 nm. The amount of total phenolics and chlorogenic acid in the fruits decreased during storage; however, the differences were not significant. The browning potential closely correlated with the enzyme activity, but not with the phenolic content.     

OXIDATIVE ENZYME ACTIVITIES AND SOLUBLE PROTEIN CONTENT IN LEAVES AND FRUITS OF OLIVES DURING RIPENING

Changes in the content and composition of soluble proteins and the activities of ionically bound peroxidase and polyphenoloxidase in leaves and fruits of olive trees Olea europaea cv. Zard from three regions during “on” and “off” years were evaluated. It was shown that peroxidase and polyphenoloxidase activities were low at early fruit developmental stages, gradually increased and were the highest at full fruit development (120–135 days after fruit set), and remained relatively high throughout ripening and softening stage. Polyacrylamide gel electrophoresis was used for characterization of peroxidase and polyphenoloxidase enzymes in leaf and fruit extracts. Isoenzymes were assessed in extracts from different regions and it was observed that some isoenzymes were only present at full fruit maturity. The protein content and its expression was estimated and characterized, and it was shown that protein content increased during various stages of fruit growth and remained the highest at full fruit ripening. The increase in enzyme activity from leaves and fruits was accompanied by differential expression of isoenzymes and the protein bands between 20 to 37 KDa, that seems to be a marker of full fruit maturity. It was also concluded that in all cases protein content and enzyme activities in leaves and fruits of olive trees during “on” years are considerably lower than those during “off” years.