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1.
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.  相似文献   

2.
Flame Seedless grapes were sprayed with N-(2-chloro-4-pyridyl)-N-phenylurea (CPPU) at 0, 2.5, and 5.0 ppm to develop rachis resistant to browning and dehydration. Rachis polyphenol oxidase (PPO) activity was determined during cluster development. Cluster components were weighed at commercial (CM), and physiological maturity (PM). PPO activity, rachis color changes (L and a), and cluster weight loss were evaluated at 0 degrees C for 8, 16, 32, and 56 days. CPPU-treated rachis had a decrease of 36% in PPO activity and a week delay in peak activity. At PM, dry weight of CPPU-treated rachis increased by 3 g. Postharvest rachis PPO activity declined with CPPU application, and color changes followed the same pattern for CM and PM. After 32 days of storage, L and a in lateral branches were significantly superior in CPPU treatments. Weight losses below 2.1% were significantly lowest in CPPU-treated clusters for 16 days of storage regardless of cluster maturity.  相似文献   

3.
The browning of glucose-fructose-glycine mixtures involves parallel glucose-glycine and fructose-glycine reactions, which share a common intermediate, the immediate precursor of melanoidins in the kinetic model. At pH 5.5, 55 degrees C glucose is converted into this intermediate in a two step process where k(1) = (7.8 +/- 1.1) x 10(-)(4) mol L(-)(1) h(-)(1) and k(2) = (1.84 +/- 0.31) x 10(-)(3) h(-)(1) according to established kinetics, whereas fructose is converted into this intermediate in a single step where k(4) = 5.32 x 10(-)(5)()()mol L(-)(1) h(-)(1). The intermediate is converted to melanoidins in a single rate limiting process where k(mix) = 0.0177 h(-)(1) and the molar extinction coefficient (based on the concentration of sugar converted) of the melanoidins so formed is 1073 +/- 4 mol(-)(1) L cm(-)(1). Whereas the value of k(mix) is the same when the individual sugars undergo browning, the value of the molar extinction coefficient is similar to that for melanoidins from the glucose-glycine reaction (955 +/- 45 mol(-)(1) L cm(-)(1)) but it is approximately double the value for melanoidins from the fructose-glycine reaction (478 +/- 18 mol(-)(1) L cm(-)(1)). This is the reason that the effects of glucose and fructose on the rate of browning are synergistic.  相似文献   

4.
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.  相似文献   

5.
超高压联合高密度CO2处理钝化对虾多酚氧化酶   总被引：1，自引：1，他引：0  
为了弥补超高压(UHP,ultra high pressure)钝化凡纳滨对虾多酚氧化酶(PPO,polyphenol oxidase)效果差的缺点,同时利用高密度CO_2钝化凡纳滨对虾PPO的优势,初步研究UHP+CO_2处理对凡纳滨对虾PPO的钝化效果,以探讨UHP+CO_2联合处理用于开发虾类新产品的可行性。研究结果表明:UHP+CO_2联合处理比单独CO_2处理和UHP处理更能有效地钝化PPO;100 MPa UHP+CO_2联合处理30 min,PPO相对酶活降至18.92%±1.52%;200 MPa UHP+CO_2联合处理10 min,PPO相对酶活降至10.91%±1.08%;300 MPa UHP+CO_2联合处理10 min,PPO被钝化95%;400 MPa UHP+CO联合处理5 min,PPO被钝化97%;500 MPa UHP+CO联合处理10 min,PPO100%被钝化;与单独UHP处理相比,UHP+CO_2联合缩短了处理时间,提高了钝化PPO的效果;PPO经UHP+CO_2联合处理后在4℃贮藏6 d后活性未见恢复,说明PPO在处理过程中发生了不可逆的变性失活。研究结果为虾类的贮藏和加工以及开发新产品提供基础数据和技术参考。  相似文献   

6.
Dominga grape polyphenol oxidase (PPO) was extracted using phase partitioning with Triton X-114. The enzyme was obtained in latent state and could be optimally activated by the presence of 0.2% sodium dodecyl sulfate (SDS) at pH 6.0. In the absence of SDS, the enzyme showed maximum activity at acid pH. The kinetic parameters of the enzyme at pH 3.0 and 6.0 in the presence of SDS were calculated. The effect of several inhibitors was studied, tropolone being the most effective with a K(i) value of 18 muM. The effect of cyclodextrins was also studied, and the complexation constant K(c) between G(2)-beta-cyclodextrins and 4-tert-butylcatechol was calculated using the enzymatic method (K(c) = 13960 M(-)(1)). The evolution of the color parameters (L, a, b) of liquefied grape berries was inhibited by inhibitors of PPO activity, such as diethyldithiocarbamate, metabisulfite, and G(2)-beta-cyclodextrins, indicating that enzymatic browning by PPO is the main process involved in the browning of Dominga grape juice at room temperature.  相似文献   

7.
CaCl_2和6-BA溶液处理对枣果软化褐变及相关酶活性的影响   总被引：2，自引：0，他引：2  
试验探讨了鲜枣贮藏过程中果肉软化褐变与 PG活性、PPO活性的关系以及采前 Ca Cl2 和 6 - BA溶液处理对采后枣果果肉软化褐变、PG活性、PPO活性的影响。结果表明 ,枣果采后硬度逐渐下降且与贮藏时间呈极显著负相关 ,PG活性在贮藏期间呈起伏变化 ,与枣果软化无显著相关性。随贮藏时间延长果肉褐变指数逐渐增大 ,PPO活性逐渐升高 ,PPO活性与果肉软化褐变的发生相一致。采前用 1% Ca Cl2 、15 mg/kg6 - BA以及 1% Ca Cl2 +15mg/kg6 - BA对枣果进行喷雾处理 ,能降低采后枣果的 PG活性和 PPO活性 ,抑制果肉硬度的下降和果肉的褐变进程。 1% Ca Cl2 处理可使赞皇大枣贮藏末期的果肉硬度比对照增加 1.2 kg/cm2 ;1% Ca Cl2 +15 m g/kg6 - BA处理可使赞皇大枣贮藏末期的果肉硬度增加 1.5 kg/cm2 。 15 m g/kg 6 - BA处理对枣果软化褐变及 PG活性、PPO活性有一定影响 ,但效果不及 Ca Cl2 处理  相似文献   

8.
Oxidation of the flavonol quercetin by polyphenol oxidase   总被引：5，自引：0，他引：5  
Because direct oxidation of flavonols by polyphenol oxidase (PPO) has not previously been reported and, given the importance of flavonols, the ability of broad bean seed PPO to oxidize the flavonol quercetin was studied. The reaction was followed by recording spectral changes with time. Maximal spectral changes were observed at 291 nm (increase) and at 372 nm (decrease). The presence of two isosbectic points (at 272 and 342 nm) suggested the formation of only one absorbent product. These spectral changes were not observed in the absence of PPO. The oxidation rate, which varied with pH, was highest at pH 5.0. The following kinetic parameters were also determined: V(m) = 11 microM/min, K(m) = 646 microM, V(m)/K(m) = 17 x 10(-)(2) min(-)(1). Flavonol oxidation was efficiently inhibited (K(I) = 3.5 microM) by specific PPO inhibitors such as 4-hexylresorcinol. The results obtained showed that quercetin oxidation was strictly dependent on the presence of PPO.  相似文献   

9.
In this study the catecholase and cresolase activities of eggplant polyphenol oxidase (PPO) were investigated. Enzyme activity was determined by measuring the increase in absorbance using catechol as substrate and 3-methyl-2-benzothiazolinone hydrazone (MBTH) as coupled reagent. The effects of substrate specificity, heat inactivation, temperature, pH, and inhibitors were investigated to understand the enzymatic alteration of ready-to-eat preparations. Browning of vegetables was determined through a colorimeter. Decrease of lightness (L*) and increase of color difference values (ΔE*) were correlated with tissue browning. Antibrowning agents were tested on PPO under the same conditions. The enzyme activity was strongly inhibited by 0.4 M citric acid. Under natural pH conditions, the enzyme was also inhibited by tartaric acid and acetic acid. All of the results were used to understand the best conditions for food transformation (ready-to-eat and grilled eggplant slices).  相似文献   

10.
封闭系统水稻土甲烷氧化的模拟   总被引：1，自引：0，他引：1  
Methane oxidation by paddy soils in a closed system could be simulated by the equation x=k1xo/(k1 k2x0)exp(k2t)-k2x0 where x0 and x are the CH4 concentrations at time zero and t,respectively;k1 and k2 are constants related to the constant of first-order-kinetics.According to the equation the change of soil ability to oxidize CH4 could be estimated by the equation Ac=k2/k1(x0-x)x0k2/k1-1.The results showed that the soil ability to oxidize CH4 varied,depending on the initial CH4 concentration.High initial CH4 concentration stimulated soil ability to consume CH4,while low concentration depressed the ablility.This characteristic of paddy soil seemed to be of considerable significance to self-adjusting CH4 emission from flooded rice fields if there exist oxic microsites in the soil.  相似文献   

11.
外源铜对土壤果树系统中酶活性影响的研究   总被引：10，自引：0，他引：10  
利用褐土和红富士苹果嫁接苗为供试材料 ,研究外源铜对土壤—苹果树系统中酶活性的影响。结果表明 ,低量的外源铜能使果树叶片的过氧化氢酶、多酚氧化酶、抗坏血酸氧化酶及根系过氧化氢酶的活性加强 ,而高量铜使活性大幅度降低 ,加入钙铁后在一定程度上使抑制缓解。土壤过氧化氢酶的活性与外源铜量具显着的曲线相关性 ,(y =x ( - 1 1 .2 6 0 .75x) ,r=0 .995 5 ) ,蔗糖酶活性与土壤施铜量呈Logistic函数关系 (y =2 .95 ( 1 0 .5 7exp( - 5 .0 2× 1 0 - 5x) ,r=- 0 .982 0 )。脲酶对铜过量非常敏感 ,大于 1 0 0mgkg- 1的各处理均未测出其活性  相似文献   

12.
To enhance the shelf life of edible mature mushrooms, Agaricus bisporus, 2 kGy ionizing treatments were applied at two different dose rates: 4.5 kGy/h (I(-)) and 32 kGy/h (I(+)). Both I(+) and I(-) showed a 2 and 4 day shelf-life enhancement compared to the control (C). Before day 9, no significant difference (p>0.05) in L value was detected in irradiated mushrooms. However, after day 9, the highest observed L value (whiteness) was obtained for the mushrooms irradiated in I(-). Analyses of phenolic compounds revealed that mushrooms in I(-) contained more phenols than I(+) and C, the latter containing the lower level of phenols. The fluctuation of the precursors of glutaminyl-4-hydroxyaniline (GHB) was less in I(-) than in I(+). The polyphenol oxidase (PPO) activities of irradiated mushrooms, analyzed via catechol oxidase, dopa oxidase, and tyrosine hydroxylase substrates, were found to be significantly lowered (p = 0.05) compared to C, with a further decrease in I(+). Analyses of the enzymes indicated that PPO activity was lower in I(+), contrasting with its lower phenols concentration. The observation of mushrooms' cellular membranes, by electronic microscopy, revealed a better preserved integrity in I(-) than in I(+). It is thus assumed that the browning effect observed in I(+) was caused by both the decompartmentation of vacuolar phenol and the entry of molecular oxygen into the cell cytoplasm. The synergetic effect of the residual active PPO and the molecular oxygen, in contact with the phenols, allowed an increased oxidation rate and, therefore, a more pronounced browning I(+) than in I(-).  相似文献   

13.
微酸性电解水结合壳聚糖对水蜜桃护色保鲜的效果   总被引：8，自引：2，他引：6  
为了防止水蜜桃(Prunus persicaL.Batsch)采后的果肉颜色变化而严重影响其商品价值。该研究以南汇水蜜桃为材料,分别利用自制羧甲基壳聚糖复合保鲜剂,以及微酸性电解水对水蜜桃进行护色保鲜处理,并对冷藏条件下(2℃)水蜜桃果肉颜色及与颜色品质变化相关的指标进行了检测,同时还利用核磁共振成像(MRI)合成T2-MAP图研究了冷藏开始和结束后水蜜桃内部变化的规律。结果表明,在贮藏结束后,对照组果肉颜色的L*值、H*值、C*值分别是羧甲基壳聚糖复合保鲜剂和微酸性电解水处理组的85%～91%、85%～93%和114%～122%(p<0.05)。显然,羧甲基壳聚糖复合保鲜剂和微酸性电解水由于可以抑制水果自身乙烯的产生,减少水果的细胞膜透性和T2-MAP图中T2弛豫常数的变化,降低了多酚氧化酶(PPO)活性的增加速度,减少了酚类物质的累积,进而延缓了果肉的变色。另外,与仅用电解水处理相比,结合羧甲基壳聚糖处理具有较好的护色保鲜效果。通过该研究,可以为提高采后南汇水蜜桃的果肉护色保鲜效果提供参考。  相似文献   

14.
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.  相似文献   

15.
苹果液氮降温排氧打浆技术研究   总被引：2，自引：0，他引：2  
为了抑制苹果制汁过程中的褐变，提高产品质量，该文首次采用自行设计的液氮降温排氧打浆机，对苹果打浆工艺进行了初步研究。具体研究了打浆时添加液氮后苹果浆温度、溶解氧含量、果浆颜色以及多酚氧化酶(PPO)和过氧化物酶(POD)活力等的变化。结果表明：添加1%～2%液氮后，苹果浆温度迅速降低为原来温度的50%，氧气含量降低为对比组的47.7%；果浆色泽L值为53.96，较对比组的L值高44.1％；同时可减少维生素C损失，提高苹果出汁率。  相似文献   

16.
The rate of consumption of dissolved oxygen by apple polyphenol oxidase in cider apple juices did not correlate with polyphenol oxidase activity in the fruits and decreased faster than could be explained by the decrease of its polyphenolic substrates. The kinetics parameters of a crude polyphenol oxidase extract, prepared from apple (Braeburn cultivar), were determined using caffeoylquinic acid as a substrate. Three apple procyanidin fractions of n 80, 10.5, and 4 were purified from the parenchyma of cider apples of various cultivars. Procyanidins, caffeoylquinic acid, (-)-epicatechin, and a mixture of caffeoylquinic acid and (-)-epicatechin were oxidized by reaction with caffeoylquinic acid o-quinone in order to form oxidation products. All the fractions were evaluated for their inhibitory effect on PPO activity. Native procyanidins inhibited polyphenol oxidase activity, the inhibition intensity increasing with n. The polyphenol oxidase activity decreased by 50% for 0.026 g/L of the fraction of n 80, 0.17 g/L of the fraction of n 10.5, and 1 g/L of the fraction of n 4. The inhibitory effect of oxidized procyanidins was twice that of native procyanidins. Oxidation products of caffeoylquinic acid and (-)-epicatechin also inhibited polyphenol oxidase.  相似文献   

17.
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.  相似文献   

18.
Polyphenol oxidase (PPO) from eggplant was extracted and partially purified by a two-step fractionation-precipitation using ammonium sulfate and phenylsepharose hydrophobic interaction chromatography. The eggplant PPO extract was characterized concerning its kinetic properties. Optimal conditions to obtain Maillard reaction products (MRPs) with a maximal inhibitory potency (IP) toward PPO activity were determined using the surface response methodology and a four-factor and five-level experimental design. The MRPs were prepared from cysteine (0.25 M) and glucose (0-1 M), at several initial pH values (2-6) and at differing heating times (3-19 h) and temperatures (95-115 degrees C). The maximal IP was obtained after heating a model system of glucose/cysteine (1/0.25 M) at pH 2 for 3 h 20 min at 115 degrees C. The soluble part of this MRP, called MRP(IPmax), was a noncompetitive inhibitor toward eggplant PPO. The IP of MRP(IPmax) on PPO activity was very potent as compared to that displayed by benzoic, p-coumaric, and t-cinnamic acids, as well as sorbic acid and 4-hexylresorcinol. The activity of preincubated PPO at 0 degrees C with MRP(IPmax) was only slightly restored after dialysis or gel filtration.  相似文献   

19.
The kinetics of the activation process of latent peach PPO by trypsin was studied. By coupling this activation process to the oxidation of 4-tert-butylcatechol (TBC) to its corresponding quinone, it was possible to evaluate the specific rate constant of active PPO formation, k(3), which showed a value of 0.04 s(-1). This proteolytic activation of latent peach PPO permitted us to characterize the monophenolase activity of peach PPO for the first time using p-cresol as substrate, and it showed the characteristic lag period of the kinetic mechanism of monophenols hydroxylation, which depended on the enzyme and substrate concentration, the pH and the presence of catalytic amounts of o-diphenol (4-methylcatechol). The enzyme activation constant, k(act), was 2 microM.  相似文献   

20.
The inactivation of apple pectin methylesterase (PME) with dense phase carbon dioxide (DPCD) combined with temperatures (35-55 degrees C) is investigated. DPCD increases the susceptibility of apple PME to the temperatures and the pressures have a noticeable effect on apple PME activity. A labile and stable fraction of apple PME is present and the inactivation kinetics of apple PME by DPCD is adequately described by a two-fraction model. The kinetic rate constants k L and k S of labile and stable fractions are 0.890 and 0.039 min (-1), and the decimal reduction times D L and D S are 2.59 and 58.70 min at 30 MPa and 55 degrees C. Z T representing temperature increase needed for a 90% reduction of the D value and the activation energy E a of the labile fraction at 30 MPa is 22.32 degrees C and 86.88 kJ /mol, its Z P representing pressure increase needed for a 90% reduction of the D value and the activation volume V a at 55 degrees C is 21.75 MPa and -288.38 cm (3)/mol. The residual activity of apple PME after DPCD exhibits no reduction or reactivation for 4 weeks at 4 degrees C.  相似文献   

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