Partial characterization of polyphenol oxidase activity in raspberry fruits.

A partial characterization of polyphenol oxidase (PPO) activity in raspberry fruits is described. Two early cultivars harvested in May/June (Heritage and Autumm Bliss) and two late cultivars harvested in October-November (Ceva and Rubi) were analyzed for PPO activity. Stable and highly active PPO extracts were obtained using insoluble poly(vinylpyrrolidone) (PVP) and Triton X-100 in sodium phosphate, pH 7.0 buffer. Polyacrylamide gel electrophoresis of raspberry extracts under nondenaturing conditions resolved in one band (R(f)()(1) = 0.25). Raspberry PPO activity has pH optima of 8.0 and 5.5, both with catechol (0.1 M). Maximum activity was with D-catechin (catecholase activity), followed by p-coumaric acid (cresolase activity). Heritage raspberry also showed PPO activity toward 4-methylcatechol. Ceva and Autumm Bliss raspberries showed the higher PPO activity using catechol as substrate.     

Characterization and role of polyphenol oxidase and peroxidase in browning of fresh-cut melon

Polyphenol oxidase (PPO) and peroxidase (POD) were extracted from two different varieties of melon ( Cucumis melo L. cantalupensis cv. Charentais and C. melo L. inodorus cv. Amarillo) and characterized using reliable spectrophotometric methods. In both cases the enzymes followed Michaelis-Menten kinetics, showing different values of kinetics parameters between the two cultivars: K m = 7.18 +/- 0.70 mM ('Charentais') and 6.66 +/- 0.20 mM ('Amarillo') mM; V max = 7.93 +/- 0.35 units/min ('Charentais') and 13.82 +/- 0.37 units/min ('Amarillo'), relative to PPO; K m = 24.0 +/- 2.10 mM ('Charentais') and 5.05 +/- 0.19 mM ('Amarillo') mM; V max = 344.83 +/- 10.32 units/min ('Charentais') and 80.64 +/- 2.01 units/min ('Amarillo'), relative to POD. Optimum pH for PPO was 7.0 for 'Charentais' and 7.5 for 'Amarillo, whereas it was 4.5 for both cultivars relative to POD. Melon PPO had maximum activity at 60 degrees C in both 'Charentais' and 'Amarillo' cultivars, whereas POD maximum activity was found at 45 degrees C in 'Charentais' and at 25 degrees C in 'Amarillo'. POD from both cultivars showed higher thermolability compared with PPO, losing >90% of relative activity after only 5 min of incubation at 70 degrees C. POD's activation energy was much higher than that of PPO (Delta E (#) = 86.3 and 160.6 kJ mol (-1) for 'Charentais' and 'Amarillo', respectively). PPO and POD activities from both cultivars showed a decreasing pattern as sugar concentration in the assay medium increased, except in POD extract from 'Charentais', which maintained its activity in the presence of high d-glucose concentration (up to 5 M). Changes in L*, a*, b*, chroma, and hue angle values were chosen to describe the browning development in the samples during storage at 5 degrees C. A slight decrease in L* value and a more marked reduction of a* value were noted in both cultivars above all at the end of storage period. POD activity during storage at 5 degrees C was highly correlated with changes of parameters a*, b*, chroma, and hue angle ( r (2) from 0.82 to 0.97) for cultivar 'Charentais'. According to these results, only POD activity seemed to be involved in browning of minimally processed melon.     

Effect of wounding on phenolic enzymes in six minimally processed lettuce cultivars upon storage

The effect of wounding on polyphenol oxidase (PPO), peroxidase (POD), and phenylalanine ammonia-lyase (PAL) was studied in six minimally processed lettuce (Lactuca sativa L.) cultivars upon storage for 7 days at 5 degrees C (Iceberg Mikonos (IM), I. Green Queen (IGQ), I. Asdrúbal (IA), Little Gem Sandra (LGS), Romaine Cazorla (RC), and R. Modelo (RM)). Wounding of lettuce tissue midribs (because of minimal processing) caused an exponential increase in PPO activity due to the activation process from latent to fully active PPO by following first order kinetics in the time range from 3.7 days (LGS) to 6.3 days (RC). However, total PPO activity (active plus latent) remained constant. Isoform pattern of PPO changed upon storage probably because of posttranslational processes. POD activity linearly increased with induction of new POD isoenzymes. PAL activity presented a typical bell-shaped induction pattern in four cultivars. Only IM and IGQ showed a second induction response which has not been previously described in the literature. IM was the cultivar most susceptible to browning and RC was the cultivar least susceptible. However, no clear correlation was observed between browning and any of the biochemical and physiological attributes investigated (PPO, PAL, and POD activities, total and individual phenols accumulation, and ascorbic acid content).     

Characterization of polyphenol oxidase and peroxidase and influence on browning of cold stored strawberry fruit

Polyphenol oxidase and peroxidase were extracted from two different varieties of strawberry fruit (Fragaria x ananassa D, cv. 'Elsanta' and Fragaria vesca L, cv. 'Madame Moutot') and characterized using reliable spectrophotometric methods. In all cases, the enzymes followed Michaelis-Menten kinetics, showing different values of peroxidase kinetics parameters between the two cultivars: Km = 50.68 +/- 2.42 mM ('Elsanta') and 18.18 +/- 8.79 mM ('Madame Moutot') mM and Vmax = 0.14 +/- 0.03 U/g ('Elsanta') and 0.05 +/- 0.01 U/g ('Madame Moutot'). The physiological pH of fruit at the red ripe stage negatively affected the expression of both oxidases, except polyphenol oxidase from 'Madame Moutot' that showed the highest residual activity (68% of the maximum). Peroxidase from both cultivars was much more thermolable as compared with PPO, losing over 60% of relative activity already after 60 min of incubation at 40 degrees C. The POD activation energy was much lower than the PPO activation energy (DeltaE = 97.5 and 57.8 kJ mol-1 for 'Elsanta' and 'Madame Moutot', respectively). Results obtained from d-glucose and d-fructose inhibition tests evidenced a decreasing course of PPO and POD activities from both cultivars as the sugar concentration in the assay medium increased. Changes in CIE L*, a*, b*, chroma, and hue angle values were taken as a browning index of the samples during storage at 4 degrees C. A decrease in L* was evident in both cultivars but more marked in 'Elsanta'. PPO and POD activities from cv. 'Elsanta' were very well-correlated with the parameter L* (r2=0.86 and 0.89, respectively) and hue angle (r2=0.85 and 0.93, respectively). According to these results, the browning of the fruit seemed to be in relation to both oxidase activities.     

Purification and characterization of polyphenol oxidase from cauliflower (Brassica oleracea L.)

Polyphenol oxidase (PPO) of cauliflower was purified to 282-fold with a recovery rate of 8.1%, using phloroglucinol as a substrate. The enzyme appeared as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The estimated molecular weight of the enzyme was 60 and 54 kDa by SDS-PAGE and gel filtration, respectively. The purified enzyme, called phloroglucinol oxidase (PhO), oxidized phloroglucinol (K(m) = 3.3 mM) and phloroglucinolcarboxylic acid. The enzyme also had peroxidase (POD) activity. At the final step, the activity of purified cauliflower POD was 110-fold with a recovery rate of 3.2%. The PhO and POD showed the highest activity at pH 8.0 and 4.0 and were stable in the pH range of 3.0-11.0 and 5.0-8.0 at 5 °C for 20 h, respectively. The optimum temperature was 55 °C for PhO and 20 °C for POD. The most effective inhibitor for PhO was sodium diethyldithiocarbamate at 10 mM (IC(50) = 0.64 and K(i) = 0.15 mM), and the most effective inhibitor for POD was potassium cyanide at 1.0 mM (IC(50) = 0.03 and K(i) = 29 μM).     

植物防御酶与桉树对焦枯病抗性的关系

为筛选鉴定桉树对焦枯病(CylindrocladiumqulnqueSeptatumMorgan)抗性的生化指标,对福建省11个桉树主栽种系接种焦枯病菌前后关键防御酶系中的过氧化物酶(POD)和超氧岐化酶(SOD)活性及其同工酶变化进行了研究.结果表明:无论是桉树健叶还是接种后的病叶,体内POD、SOD活性均表现为抗病种系>中抗种系>中感种系>感病种系,呈规律性变化;接种前后POD同工酶谱带数与桉树对焦枯病的抗性间不呈规律性变化,而SOD谱带数与抗病性成正比.从抗病育种角度出发可将接种前后桉树POD、SOD活性和接种后SOD同工酶谱带作为桉树对焦枯病抗性早期鉴定的生化指标.     

莲藕膨大过程中褐变酶活性的变化

本实验以3个熟性不同的莲藕品种为材料,研究了莲藕膨大过程中褐变酶活性的变化。实验表明:随着膨大节数的增加,过氧化物酶(POD)、多酚氧化酶(PPO)的活性增加;第4节藕体随着采收时间的推迟,褐变酶活性增加,且熟性不同品种之间有明显差异。并探讨了品种熟性、藕体大小和环境条件对莲藕褐变酶的影响。     

1-MCP负压渗透处理对鲜枣贮藏期间生理及抗氧化能力的影响

以山东大瓜枣为试材,研究1-甲基环丙烯（1-MCP）负压渗透处理对鲜枣贮藏期间生理指标及抗氧化能力的影响。结果表明：1-MCP处理可以维持鲜枣果实体内较低的活性氧水平（H2O2）,提高果实自身抗氧化能力（T-AOC、OH·、O2·-、DPPH）,维持果实自身的抗氧化物质抗坏血酸（AsA）及总酚含量;1-MCP处理对果实过氧化氢酶（CAT）呈现抑制表达作用,而对于抗坏血酸过氧化物酶（APX）、超氧化物歧化酶（SOD）、多酚氧化酶（PPO）和过氧化物酶（POD）则有一定的激活作用。     

Phenolic compounds and related enzymes are not rate-limiting in browning development of fresh-cut potatoes

The effect of minimal processing on polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia-lyase (PAL), and phenolic compounds was studied in five potato cultivars (Agria, Cara, Liseta, Monalisa, and Spunta). Minimal processing caused an overall increase in PPO, POD, and PAL activities. The isoform pattern of PPO was the same for all of the cultivars before and after processing. No latent PPO was detected. The isoperoxidase pattern was approximately the same among cultivars. An increase in POD activity was related to the specific induction of an acidic isoperoxidase. PAL showed an induction pattern characterized by the presence of a maximum peak of activity after 4 days of processing for all of the cultivars. The sequence of browning susceptibility of potato cultivars was as follows: Monalisa > Spunta > Liseta > Cara > Agria. Browning development was only partially correlated to PAL activity (only during the first 4 days after wounding). However, this correlation could not explain the above sequence of browning susceptibility. Minimal processing caused an increase of chlorogenic acid, whereas tyrosine content remained unchanged. In summary, no significant correlation was found between either rate or degree of browning and any other biochemical and physiological attribute investigated (PPO, POD, hydrogen peroxide, ascorbic acid content, and initial phenolics content as well as total and individual phenolics accumulation).     

Polyphenol oxidase and peroxidase expression in four pineapple varieties (Ananas comosus L.) after a chilling injury

Pineapple internal browning (IB) is a chilling injury that produces enzymatic browning associated with flesh translucency. Pineapple biodiversity allowed the investigation of how polyphenol oxidase (PPO) and peroxidase (POD) activities with their different isoforms are involved in the IB mechanism. Fruits of four varieties that expressed IB symptoms differently, Smooth Cayenne (SCay) and the hybrids MD2, Flhoran 41 (Flh 41), and Flhoran 53 (Flh 53), were stressed by cold. The susceptible varieties showed classical brown spots but different patterns of IB, whereas MD2 and controls showed no IB. Enzymatic activities were measured on fruit protein extracts and PPO and POD isoforms separated on mini-gels (PhastSystem). Only PPO activity was significantly enhanced in the presence of IB. Up to six PPO isoforms were identified in the susceptible varieties. PPO was barely detectable in the nonsusceptible variety MD2 and in controls. The number of PPO isoforms and the total PPO activity after chilling are varietal characteristics.     

Activity and Inhibition of Polyphenol Oxidase in Extracts of Bran and Other Milling Fractions from a Variety of Wheat Cultivars

Studies were conducted to compare polyphenol oxidase (PPO) specific activities in various milling fractions of a variety of wheat cultivars and determine the levels of activities in a number of cultivars from different localities and harvesting seasons. Substrate specificities were also investigated. Bran was singled out as the richest source of PPO activity, which may also influence the activity in the other milling fractions that are known to have some proportion of bran content. We showed by gel electrophoresis and spectrophotometrically that the protein responsible for PPO activity apparently exists as a single isoform in bran and that the observed enzyme activity is likely to be a tyrosinase type, not a laccase or peroxidase. The specific activity was not significantly different between the reduction shorts and break shorts from the same cultivar, indicating a similar level of bran contamination in these fractions. Very low levels of PPO activity were recorded in the flour of all cultivars studied. Bran was used, therefore, to determine the varietal differences in the PPO activities in a number of cultivars from different localities and seasons of harvest. Results showed that the most significant determinant of PPO activity was the genotype, and this may be influenced by seasonality. We also determined that, apart from substrate preferences by the PPO enzyme, some phenolic acids actually inhibit PPO. Furthermore, we found that bran of some cultivars extracted with acidified methanol inhibited PPO activity substantially, whereas other extracts had less inhibitory properties. Thus, these unknown compounds in wheat may inhibit endogenous PPO activity.     

涂膜和热处理对葡萄能量和贮藏生理及品质的影响

研究不同处理对欧亚种"维多利亚"葡萄组织能量水平和生理品质的影响。采后葡萄经钙联合涂膜和热处理,即在质量分数为0.2%Ca Cl2和1%壳聚糖中浸泡20 min,45℃的0.2%Ca Cl2溶液中浸泡20 min,晾干后(4±0.5)℃贮藏,每10d测定果实的呼吸强度、褐变指数(browning index,BI)、腐烂率、硬度、丙二醛(malondialdehyde,MDA)含量、三磷酸腺苷(adenosine triphosphate,ATP)、二磷酸腺苷(adenosine diphosphate,ADP)、单磷酸腺苷(adenosine monophosphate,AMP)的含量,多酚氧化酶(polyphenol oxidase,PPO)、过氧化物酶(peroxidase,POD)、脂氧合酶(lipoxygenase,LOX)和超氧化物歧化酶(soperoxide dismutase,SOD)活性变化,以未处理为对照。结果显示对照组葡萄组织中能量物质处于亏损状态,呼吸强度减弱,细胞膜氧化增加,膜透性、BI、腐烂率增加,PPO、POD和LOX活性增加,SOD降低,果实软化。钙联合热和涂膜处理较好维持了果实的能量水平和生理品质,且涂膜优于热处理。ATP与硬度、SOD显著正相关(r=0.938,0.930,P0.01),与MDA、膜透性、LOX显著负相关(r=-0.896,-0.932,-0.940,P0.01);能荷(energy charge,EC)值与膜透性、LOX负相关(P0.05),与呼吸强度和SOD活性正相关(P0.05)。随贮藏时间的延长,葡萄组织的能量水平处于亏损状态,能量水平显著影响生理品质。经适当的采后处理可延缓能量亏损,涂膜优于热处理。     

Polyphenol Oxidase in Wheat Grain: Whole Kernel and Bran Assays for Total and Soluble Activity

Color is a key quality trait of wheat products, and polyphenol oxidase (PPO) is implicated as playing a significant role in darkening and discoloration. In this study, total and soluble PPO activities were characterized in whole kernel assays and bran extracts. In whole kernel assays similar to AACC Approved Method 22–85, four wheat cultivars were ranked the same for both total and soluble (leached) PPO activity with L‐DOPA (diphenol) as the substrate. Total kernel PPO activity was much greater than soluble PPO activity in three hexaploid wheat cultivars, indicating that insoluble PPO was the major contributor to kernel PPO measurements. Tyrosine (monophenol) was an excellent PPO substrate in kernel assays as expected but had no activity as a substrate for soluble PPO. However, soluble PPO activity with tyrosine was activated by the addition of the diphenols chlorogenic acid and caffeic acid. When PPO was assayed in homogenized bran, 89–95% of total PPO activity remained insoluble, associated with the bran particles. The kernel assay detected <2% of PPO measured in an equivalent amount of homogenized bran. However, total PPO activity was 2‐fold higher in Klasic than in ID377s, both when measured in the kernel assay and in homogenized bran, indicating that the kernel assay was an accurate predictor of relative total extracted PPO activity in these two cultivars. Adding detergents (0.1% SDS plus 0.2% NP‐40) to the bran extraction buffer increased both soluble and insoluble PPO activity. Results indicate that relative PPO activities among wheat cultivars are similar in whole kernel and kernel leachate assays, and that the predominant insoluble fraction of PPO, which is relatively uncharacterized, may be largely responsible for wheat product discoloration.     

石榴果实发育期果皮褐变及相关酶活性变化

为探明石榴果实发育期果皮褐变及其相关酶活性、总酚和抗氧化能力的变化规律及相互关系,以泰山红和泰山三白甜石榴为试材,研究各发育期果皮褐变度、多酚氧化酶(PPO)活性、过氧化物酶(POD)活性、苯丙氨酸解氨酶(PAL)活性、总酚含量和DPPH自由基清除率的变化。结果表明,2个品种果皮褐变度、PPO活性、POD活性、总酚含量和DPPH自由基清除率随着发育天数的增加逐渐降低,均在7月15日达到最高。2个品种果实发育期果皮PAL活性呈先降低后升高再降低的变化趋势,分别在7月15日和9月13日出现峰值。泰山红抗褐变能力高于泰山三白甜,二者分别在9月23日和9月13日采收能减轻褐变的发生。相关性分析表明,PAL、PPO、POD、总酚和DPPH自由基清除率均与石榴果实酶促褐变密切相关,石榴果实酚类物质含量高低与其抗氧化能力极显著正相关。本研究结果为深入揭示石榴果实褐变机理提供了理论依据。     

硅对水稻防御性关键酶活性的影响及其与抗稻瘟病的关系

通过室内溶液培养试验,研究了硅酸盐对接种稻瘟病菌后水稻的病情指数、植株生物量、过氧化物酶（POD）、多酚氧化酶（PPO）和苯丙氨酸解氨酶（PAL）活性的影响。结果表明,施硅能显著降低水稻稻瘟病的发病率和病情指数,防治效果达60.59%;显著减轻稻瘟病对水稻地上部的危害,干物质量显著高于不施硅处理,但对地下部生长影响不明显;接种后水稻叶片POD活性均升高,在第5 d达到最大值,施硅处理显著低于不施硅处理,与稻瘟病抗性成负相关;水稻叶片PPO活性在接种后第5 d达到峰值,从第4 d开始施硅处理显著高于不施硅处理;接种后水稻叶片的PAL活性快速升高,在第24 h达到最大值,施硅处理显著高于不施硅处理,是其1.44倍。说明硅能提高防御性关键酶的活性,参与水稻本身防卫机制,通过一系列的生理生化作用来增强水稻的抗性。     

春石斛兰类原球茎分化过程中抗氧化酶活性和可溶性蛋白的变化

以春石斛兰(Dendrobium nobileSw.)为试材,研究了类原球茎在分化芽和根系发生过程中抗氧化酶系统中多种酶活性及可溶性蛋白的变化规律。结果表明,在分化过程中抗坏血酸过氧化物酶(APX)、愈创木酚过氧化物酶(POD)、多酚氧化酶(PPO)和过氧化氢酶(CAT)的活性和可溶性蛋白含量均呈现先降后升的趋势;超氧化物歧化酶(SOD)活性则呈现先升后降的变化。对过氧化物酶同工酶和可溶性蛋白进行SDS-PAGE电泳发现,在分化过程中均有新谱带的产生或原有谱带的消失。     

1- MCP和60Co-γ辐照处理对自发气调包装大五星枇杷贮藏品质的影响

为研究1-甲基环丙烯(1-MCP)熏蒸和⁶⁰Co-γ辐照处理对枇杷贮藏品质的影响,本研究以大五星枇杷为试验材料,对采后自发气调包装枇杷的腐烂指数、木质素、硬度、呼吸强度、乙烯生成速率、相对电导率、可滴定酸(TA)、可溶性固形物(TSS)以及苯丙氨酸解氨酶(PAL)、脂氧合酶(LOX)、多酚氧化酶(PPO)、过氧化物酶(POD)活性进行测定,研究6±0.5℃条件下1 μL·L^-1 1-MCP、1.5 kGy⁶⁰Co-γ、1 μL·L^-1 1-MCP+1.5 kGy ⁶⁰Co-γ不同处理对枇杷贮藏品质的影响。结果表明, 1-MCP和⁶⁰Co-γ复合处理效果最好,可延缓腐烂指数和木质素含量上升,维持果实硬度,降低果实呼吸强度、乙烯生成速率和相对电导率,保持果实TA和TSS含量,抑制PAL、LOX、PPO和POD活性的上升。在贮藏40 d时,1-MCP和⁶⁰Co-γ复合处理枇杷的腐烂指数仅为17.95%,而1-MCP组、⁶⁰Co-γ组、CK组分别为22.86%、25.72%和36.24%。因此,1-MCP和⁶⁰Co-γ复合处理对枇杷果实的贮藏效果最好。本研究结果为枇杷的贮藏保鲜提供了理论依据。     

Three polyphenol oxidases from red clover (Trifolium pratense) differ in enzymatic activities and activation properties

Polyphenol oxidases (PPOs) oxidize o-diphenols to o-quinones, which cause browning reactions in many wounded fruits, vegetables, and plants including the forage crop red clover (Trifolium pratense L.). Production of o-quinones in red clover inhibits postharvest proteolysis during the ensiling process. The cDNAs encoding three red clover PPOs were expressed individually in alfalfa (Medicago sativa L.), which lacks detectable endogenous foliar PPO activity and o-diphenols. Several physical and biochemical characteristics of the red clover PPOs in alfalfa extracts were determined. In transgenic alfalfa extracts, red clover PPOs exist in a latent state and are activated (10-40-fold increase in activity) by long incubations (>2 days) at ambient temperature or short incubations (<10 min) at > or =65 degrees C. PPO1 appears to be more stable at high temperatures than PPO2 or PPO3. During incubation at ambient temperature, the molecular masses of the PPO enzymes were reduced by approximately 20 kDa. The apparent pH optima of latent PPO1, PPO2, and PPO3 are 5.5, 6.9, and 5.1, respectively, and latent PPO1 is slightly activated (~5-fold) by low pH. Activation of the PPOs shifts the pH optima to approximately 7, and the activated PPOs retain substantial levels of activity as the pH increases above their optima. The latent and activated PPOs were surveyed for ability to oxidize various o-diphenols, and activation of the PPOs had little effect on substrate specificity. Activation increases the V max but not the affinity of the PPO enzymes for caffeic acid. Results indicate red clover PPOs undergo structural and kinetic changes during activation and provide new insights to their effects in postharvest physiology.     

Role of polyphenol oxidase and peroxidase in the generation of black tea theaflavins.

It has been reported earlier that when macerated tea leaf is fermented at lower pH, the resultant black tea contains increased levels of theaflavin, an important quality marker in black tea. In an attempt to investigate the biochemistry and chemistry underlying this observation, in vitro oxidation experiments using polyphenol oxidase (PPO) from fresh tea leaves, horseradish peroxidase (POD), and tea catechins, precursors for theaflavins, were carried out. In vitro oxidation experiments using crude tea PPO resulted in higher content of theaflavins at pH 4.5 in comparison with pH 5.5, the normal pH of the macerated tea leaf. When purified PPO was used in the in vitro system, surprisingly a reversal of this trend was observed, with more theaflavins being formed at the higher pH. A combination of pure tea PPO and POD led to an observation similar to that with the crude enzyme preparation, suggesting a possible role for POD in the formation or degradation of theaflavin. POD was observed to oxidize theaflavins in the presence of H(2)O(2), leading to the formation of thearubigin, another black tea pigment. This paper demonstrates that tea PPO, while oxidizing catechins, generates H(2)O(2). The amount of H(2)O(2) produced is greater at pH 5.5, the optimum pH for PPO activity, than at pH 4.5. Hence, an observed increase of theaflavins in black teas fermented at pH 4.5 appears to be due to lower turnover of formed theaflavins into thearubigins.     

Characterization of a tomato polyphenol oxidase and its role in browning and lycopene content

Polyphenol oxidase (PPO) was extracted from five Sicilian varieties of tomato fruit [Pizzutello, Naomi (Hazera), F1 PS212 (Peto seed), Rosa Maletto, and PO228] and assayed with a method using 3-methylbenzothyazolinone hydrazone (MBTH) as chromophore coupling agent. 3,4-Dihydroxyphenylacetic acid was chosen for tomato PPO activity determination. The tomato PPO had maximum activity at pH 4.8. The pH of juice in ripe fruits is between 4.1 and 4.4, a range in which PPO relative activity is between 74 and 87%. The optimum temperature of activity for tomato PPO was 40 degrees C; the enzyme showed a good relative activity (55% of the maximum) at cold-storage temperature (4 degrees C). PPO retained 82% relative activity at an NaCl concentration of 0.1 M; at higher concentrations the PPO became gradually inactivated. The commercial variety Naomi is more susceptible to enzymatic browning than the local varieties Pizzutello, Rosa Maletto and PO228, due to higher PPO activity levels. This result confirms the suitability of these local tomato varieties to national markets. Results from storage tests seem to relate PPO activity with color changes associated with browning and lycopene degradation, because lycopene is an antioxidant agent that reconstitutes the polyphenols oxidized by the action of PPO.