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.     

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.     

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).     

Polyphenol oxidase activity,phenolic acid composition and browning in cashew apple (Anacardium occidentale, L.) after processing

This study describes the extraction and characterisation of cashew apple polyphenol oxidase (PPO) and the effect of wounding on cashew apple phenolic acid composition, PPO activity and fruit browning. Purification factor was 59 at 95% (NH₄)₂SO₄ saturation. For PPO activity, the optimal substrate was catechol and the optimum pH was 6.5. PPO K_m and V_max values were 18.8 mM and 13.6 U min⁻¹ ml⁻¹, respectively. Ascorbic acid, citric acid, sodium sulphite and sodium metabisulphite decreased PPO activity, while sodium chloride increased PPO activity. Wounding at 2 °C and 27 °C for 24 h increased PPO activity but storage at 40 °C reduced PPO activity. Gallic acid, protocatechuic acid and cinnamic acid (free and conjugate) were identified in cashew apple juice. Cutting and subsequent storage at 40 °C hydrolysed cinnamic acid. 5-Hydroxymethylfurfural content in cashew apple juice increased after injury and storage at higher temperatures, indicating non-enzymatic browning.     

Identification of (−)-epicatechin as the direct substrate for polyphenol oxidase from longan fruit pericarp

Postharvest browning of longan fruit results in a short life and a reduced commercial value. The experiments were conducted to separate, then purify and finally identify the polyphenol oxidase (PPO) substrates that cause longan fruit to brown. PPO and its substrates were, respectively, extracted from longan fruit pericarp tissues. The substrate for longan PPO was separated and purified using polyamide column chromatography, Sephadex LH-20 column chromatography and silica gel column chromatography, respectively. The substrate was further identified by 0.5% FeCl₃ solution and enzymatic reaction with longan PPO. On the bases of UV, ¹H NMR, ¹³C NMR, and ESI-MS data, the direct substrate for the PPO from pericarp tissues of longan fruit was identified to be (−)-epicatechin. Furthermore, the contents of (−)-epicatechin of pericarp tissues of longan fruit of two major cultivars were determined by high performance liquid chromatography (HPLC). The HPLC analysis exhibited that the contents of (−)-epicatechin of fruit pericarp of ‘Shixia’ and ‘Chuliang’ were 0.26 and 0.56 mg/g on fresh weight (FW) basis at harvest and 0.15 and 0.09 mg/g FW after 3 days of storage. The more rapid decrease in the (−)-epicatechin content of ‘Chuliang’ was due to the oxidization catalyzed by PPO, which was in agreement with the higher browning index.     

Influences of preharvest spraying Cryptococcus laurentii combined with postharvest chitosan coating on postharvest diseases and quality of table grapes in storage

The effects of preharvest spray with Cryptococcus laurentii combined with chitosan coating after harvest on decay and quality of table grapes during storage periods were evaluated in the present study. Preharvest spray with C. laurentii (PreA) significantly decreased decay index (DI), and postharvest chitosan coating (PCC) enhanced the effectiveness of the pre-harvest spray when fruits were stored at 0 °C. PreA combination with PCC increased the activities of polyphenol oxidase (PPO) and phenylalanine ammonia-lyase (PAL) of fruit in storage. PreA + PCC treatment was effective in reducing weight loss of fruits by 85% at 17 d storage and 38% at 42 d storage as compared to PreA alone at the same stage. In addition, PreA enhanced the ratio of soluble solids content (SSC) to titratable acid (TA) by 12% at harvest time, 7% at 17 d storage and 25% at 42 d storage, mainly by increasing SSC and decreasing TA in fruit stored at 0 °C. These results suggested that integration of preharvest spray with C. laurentii and postharvest chitosan coating treatment may be a promising management strategy for decay control and quality maintenance of table grapes.     

Physiological responses and quality attributes of ‘Kyoho’ grapes to controlled atmosphere storage

This research studied the physiological responses and quality attributes of Kyoho grapes (Vitis vinifera X V. labrusca) to controlled atmosphere storage. The grapes were stored for up to 60 days in 95% relative humidity with four different conditions, 4% O₂+9% CO₂, 4% O₂+30% CO₂, 80% O_2, and air, as control. The examined physiological responses and quality attributes included polyphenol oxidase (PPO) activity, ethanol concentration, fruit detachment force (FDF), firmness, color, soluble solid content (SSC), titratable acidity (TA), ascorbic acid concentration (Vc), and sensory quality. PPO activity, FDF drop and decay incidence when stored in 4% O₂+30% CO₂ were more effectively controlled, but unacceptable alcoholic flavor and browning were detected after 45 days, compared with those stored in 4% O₂+9% CO₂ or 80% O₂. The fruits kept in 4% O₂+9% CO₂ or 80% O₂ had good quality during 60 days of storage. The results suggested that high O₂ atmosphere exhibited a potential for maintaining the quality of ‘Kyoho’ grapes during long-term storage.     

采后1-MCP和MAP处理对‘红富士’苹果冷藏和货架期品质的影响

对‘红富士’苹果进行1.0 μL/L 1-甲基环丙烯（1-methylcyclopropene,1-MCP）和自发气调包装（modified atmosphere package,MAP）处理,继而进行0 ℃冷藏和20 ℃货架贮藏。结果表明：贮藏期间,‘红富士’苹果果实硬度和可滴定酸含量下降,可溶性固形物含量升高,虎皮病和果心褐变增多。1-MCP处理能较好维持冷藏期间‘红富士’苹果果实硬度和可溶性固形物含量,降低了包装内CO2和乙烯含量。同时,1-MCP明显降低了冷藏期间虎皮病发病指数、果心褐变指数以及果柄端果肉褐变率,显著抑制果皮α-法尼烯及共轭三烯的生成。1-MCP＋MAP结合使用可较好维持果实可滴定酸含量和果皮色泽、抑制果柄端果肉褐变。综合分析认为,1-MCP＋MAP处理能较好维持‘红富士’果实冷藏和货架期间的品质,并显著抑制果实虎皮病的发生。     

1-MCP结合微孔、打孔保鲜袋对‘富士’苹果贮藏品质的影响

为了解‘富士’苹果在冷藏条件下适合的保鲜处理方式,探究了在(0±1) ℃贮藏条件下,微孔、打孔保鲜袋结合小包装便携1-MCP处理(处理剂量为0.875 μL/L)对‘富士’苹果果实硬度、外观色泽、TSS含量、TA含量、MDA含量、POD活性和PPO活性变化的影响。试验共设5个处理:纸箱衬微孔膜包装、纸箱衬微孔膜包装+1-MCP、纸箱衬打孔膜包装、纸箱衬打孔膜包装+1-MCP处理、纸箱装未加任何包装的裸果为对照(CK)。结果表明:1-MCP结合保鲜袋处理能有效延缓富士苹果果实硬度、TSS、TA的下降速度,抑制a*值降低和b*值升高,降低贮藏后期MDA的生成量,保持POD活性,并使之保持在较高水平,有效抑制PPO活性,延迟PPO活性高峰。其中以微孔+1-MCP处理效果最佳,贮藏225 d时,微孔保鲜袋结合1-MCP处理TA含量为0.12%,硬度为5.78 kg/cm2,MDA含量最低,POD活性最高,PPO活性最低,分别为9.63 mmol/g、15.47 U/(min·g)和8.82 U/(min·g),且这些指标与对照均差异显著(P<0.05)。因此,建议生产中采用微孔保鲜袋和小包装1-MCP处理剂的内包装。     

Effect of integrated application of chitosan coating and modified atmosphere packaging on overall quality retention in litchi cultivars

BACKGROUND: The storage life of litchi is limited due to pericarp browning and decay. Modified atmosphere packaging (MAP) showed promising results for ensuring quality retention. However, to improve the efficiency of MAP the integrated treatment of a chitosan coating and MAP was investigated. RESULTS: The effect of chitosan (1.0 g L⁻¹) + MAP was compared with MAP (control), and was effective in preventing decay, browning and retaining the pericarp colour in the cultivar McLean's Red. Chitosan (1.0 g L⁻¹) + MAP significantly reduced polyphenol oxidase (PPO) and peroxidase (POD) activity, retained membrane integrity, anthocyanin content and prevented the decline of pericarp colour values during storage. The POD activity was greater than the PPO activity in the cultivars McLean's Red and Mauritius. The two cultivars differed in anthocyanin content and the activity of oxidation enzymes. The gas compositions within the packages were compared between chitosan at 1.0 g L⁻¹ and 20.0 g L⁻¹ concentration for both cultivars. Chitosan (20.0 g L⁻¹) + MAP lowered the respiration during storage in both cultivars compared to 1.0 g L⁻¹ + MAP. CONCLUSION: The McLean's Red cultivar is better suited for chitosan (1.0 g L⁻¹) + MAP integrated treatment than is Mauritius in retaining overall quality. Copyright © 2009 Society of Chemical Industry     

Quality and biochemical changes of longan (Dimocarpus longan Lour cv. Daw) fruit under different controlled atmosphere conditions

Sulphur dioxide has been used to control pericarp browning in longan fruit. However, due to health and regulatory concerns, alternative treatments should be tested. The objective of this study was to find the tolerance levels of longan fruit to low O₂ (2%, 5%, 10% and 15%) and elevated CO₂ (5%, 10%, 15% and 20%) at 2 °C. According to the tolerance study, controlled atmospheres (CA) of 5% O₂ + 5% CO₂, 5% O₂ + 10% CO₂ and 5% O₂ + 15% CO₂ were compared with normal air (control) at 2 °C. Pericarp browning and decay incidence of longan were significantly (P ≤ 0.05) higher in control than all the CA treatments. CA storage reduced polyphenol oxidase (PPO) activity, maintained L* value and slowed down a decrease in total phenolic contents (TPC). Pericarp browning was highly correlated with PPO, L* and TPC.     

EFFECT OF OXYGEN CONCENTRATION ON THE BIOCHEMICAL AND CHEMICAL CHANGES OF STORED LONGAN FRUIT

ABSTRACT

Longan fruits were stored for 6 days in atmosphere of 5, 21 (air) or 60% O₂ (balance N₂) at 28C and 90–95% relative humidity to examine effects of low and high O₂ concentration on enzymatic browning and quality attributes of the fruit. Changes in pericarp browning, pulp breakdown, disease development, total phenol content, activities of phenol metabolism‐associated enzymes, relative leakage rate, α,α‐diphenyl‐β‐picrylhydrazy (DPPH) radical scavenging activity, and contents of total soluble solids, titratable acidity and ascorbic acid were evaluated. Storage of fruit in a 5% O₂ atmosphere markedly delayed pericarp browning in association with maintenance of high total phenolic content and reduced activities of polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonia lyase. Moreover, the fruit stored in a 5% O₂ atmosphere exhibited a lower relative leakage rate and higher DPPH radical scavenging activity than fruit stored in air. This presumably was beneficial in maintaining compartmentation of enzymes and substrates, and thus, reducing pericarp browning. Pulp breakdown and disease development were also reduced by exposure to a 5% oxygenatmosphere. On the contrary, exposure of longan fruit to a 60% O₂ atmosphere accelerated pericarp browning, pulp breakdown and decay development. PPO and POD activities and relative leakage rate were similar for control and 60% O₂‐treated fruit after 4 and 6 days of storage. Furthermore, treatment with 60% O₂ significantly decreased the phenolic content and DPPH scavenging activity of fruit. In addition, exposure to 5 or 60% O₂ resulted in a higher level of total soluble solids, but a lower level of ascorbic acid of longan fruit flesh. In conclusion, exposure to a 5% O₂ atmosphere showed great potential to reduce pericarp browning and extend shelf life of longan fruit.

PRACTICAL APPLICATIONS

Pericarp browning and pulp breakdown are the major causes of deterioration in postharvest longan. Conventional controlled atmosphere with low O₂ and high CO₂ is effective in maintaining quality and extending shelf life of fruits and vegetables, including inhibition of tissue browning. In this study, 5%‐controlled atmosphere reduced significantly pericarp browning, pulp breakdown and rot development. It could potentially be useful as a postharvest technology of longan fruit for reducing or replacing the use of chemicals such as SO₂ and fungicides, but it requires further investigation.     

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.     

Shelf-life extension of highbush blueberry using 1-methylcyclopropene stored under air and controlled atmosphere

The potential of 1-methylcyclopropene for controlling ripening in ‘Lateblue’ blueberry fruit was explored. After harvest, blueberry fruits were exposed to 1-MCP (0.3 and 0.6 μl l⁻¹). After treatment, samples were stored in air at 0 °C for 35 days and in a controlled atmosphere (3 kPa O₂ + 11 kPa CO₂) for 60 days. Quality parameters were monitored (weight loss, total soluble solids content, titratable acidity, firmness, anthocyanin content, phenolic content, total antioxidant capacity). Blueberries treated with 1-MCP showed a reduced weight loss during storage and a lower total soluble solid content compared to untreated fruit. High titratable acidity values were observed after controlled atmosphere storage, but no significant effect of 1-MCP on this parameter was observed. 1-MCP had no significant effects on anthocyanins, phenolics or antioxidant activities.     

Effects of aqueous chlorine dioxide treatment on browning of fresh-cut lotus root

Effect of aqueous chlorine dioxide (ClO₂) treatment on browning of fresh-cut lotus root (FLR) was investigated to explore the feasibility to apply ClO₂ for browning inhibition of fresh-cut products. Cut lotus roots were treated in ClO₂ solutions at different concentrations (10, 50 and 100 mg/l) for different time (5, 10 and 15 min), followed by chilled storage for 8-10 days at 4 °C. Color parameters (L^∗, a^∗ and b^∗), polyphenol oxidase (PPO) activity and overall visual quality (OVQ) were measured at one-day interval during storage. Results showed that higher ClO₂ concentration and longer treatment time can provide better inhibitory effects on the browning of FLR. ClO₂ concentration, treatment time and storage time were three significant factors (P < 0.05) and some significant interactions were observed. PPO activities were largely inhibited by 100 mg/l ClO₂ treatment for 10 min. The 100 mg/l ClO₂ treatment maintained high OVQ scores during 10-day storage; while 50 mg/l ClO₂ treatment was acceptable for maintaining OVQ during 4-day storage. ClO₂ treatment was demonstrated to be a promising alternative approach to control browning and improve OVQ of FLR.     

纳米乳涂膜和1-甲基环丙烯处理对枇杷果实保鲜效果的影响

以‘白砂’枇杷果实为实验材料,以清水处理为对照,研究了纳米涂膜保鲜剂（nanoemulsion coating,NC）-1处理、1-甲基环丙烯（1-methylcyclopropene,1-MCP）处理、NC-1＋1-MCP处理对枇杷在低温（4 ℃）贮藏30 d过程中品质和生理代谢的影响。结果表明：NC-1＋1-MCP处理对枇杷的贮藏品质保持效果最佳,除VC含量外,贮藏结束时,该组果实硬度、腐烂率、质量损失率都显著低于其他处理组,且NC-1＋1-MCP处理更显著抑制了枇杷在贮藏过程中苯丙氨酸解氨酶的活力,从而抑制了木质素的积累;同时抑制了多酚氧化酶和过氧化物酶的活力,从而延缓了果实的褐变衰老。研究结果表明,相比于单一的NC-1或1-MCP处理,NC-1＋1-MCP处理对枇杷保鲜效果更好。     

Effect of oxalic acid on control of postharvest browning of litchi fruit

Litchi (Litchi chinensis Sonn.) fruit, cv. Huaizhi, was treated with 2 and 4 mM oxalic acid and stored at room temperature to investigate the effect of oxalic acid on pericarp browning. The results showed that the pericarp browning indices of the fruit, treated with both oxalic acid concentrations, were significantly lower than that of the control, due to increase of membrane integrity, inhibition of anthocyanin degradation, decline of oxidation, and maintanance of relatively low peroxidase activity in the fruit during storage. It appears that application of oxalic acid can effectively control the pericarp browning of litchi fruit during postharvest storage.     

Antioxidant activities and contents of polyphenol oxidase substrates from pericarp tissues of litchi fruit

The experiments were performed to extract and purify substrates for polyphenol oxidase (PPO) from pericarp tissue of postharvest litchi fruit. Two purified PPO substrates were identified as (−)-epicatechin and procyanidin A2. The antioxidant properties of two PPO substrates were further evaluated in the present study. Variation in the content of the major substrate (−)-epicatechin of litchi fruit during storage at 25 °C was analysed using the HPLC-UV method. The results showed that (−)-epicatechin exhibited stronger antioxidant capability than procyanidin A2, in terms of reducing power and scavenging activities of DPPH radical, hydroxyl radical and superoxide radical. Furthermore, (−)-epicatechin content in pericarp tissue tended to decrease with increasing skin browning index of litchi fruit during storage at 25 °C. Thus, these two compounds can be used as potential antioxidants in litchi waste and the fresh pericarp tissue of litchi fruit exhibited a better utilisation value.     

Inactivation kinetics and secondary structural change of PEF-treated POD and PPO

Effects of pulsed electric fields (PEF) on the activity of peroxidase (POD) and polyphenol oxidase (PPO) in buffered solution were studied while the corresponding changes to their secondary structures was demonstrated by far-UV Circular dichroism (CD). The relative residual activity of POD and PPO decreased with the increase in electric field strength and treatment time, and PPO was more susceptible than POD to PEF treatment. The greatest reduction of the activity was achieved for POD at 25 kV/cm for 1740 μs and PPO at 25 kV/cm for 744 μs with reductions of 32.2% and 76.2%, respectively. The inactivation kinetic parameters D-value and Z_E value were calculated. The D-values of PPO were smaller than those POD at higher electric field strength, and Z_E values of POD and PPO were 36.9 and 16.2 kV/cm, respectively. The secondary structures of the two enzymes were changed following treatment by PEF. The intensity of negative peaks in the CD spectra decreased, and the CD spectra of PPO changed more significantly than that of POD; the reduction of the relative α-helix fractions for POD at 25 kV/cm for 124 μs was 22.63% while it was 50.72% for PPO at 25 kV/cm for 52 μs. The inactivation of PEF-treated POD and PPO was in close agreement with their secondary structure changes.