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1.
多酚氧化酶（PPO）是生物体内黑色素合成的关键酶。研究了甜柿叶乙醇提取物、涩柿叶乙醇提取物、甜柿叶水提取物、涩柿叶水提取物对马铃薯多酚氧化酶活性的抑制作用。结果表明,上述4种柿叶提取物对马铃薯多酚氧化酶具有明显的抑制作用,使该酶活力下降50％所需的浓度（IG50）分别为0．21、0．26、0．37、0．45mg／mL。乙醇提取物较水提取物的作用效果更强,甜柿叶提取物的抑制作用比涩柿叶提取物的强,4种柿叶提取物对酶的抑制作用均为非竞争性可逆抑制,其抑制常数（KI=KIS）分别为0．18、0．23、0．34、0．45mg／mL。 相似文献
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大棚菜田种植年限对土壤重金属含量及酶活性的影响 总被引：8，自引：1，他引：7
为了探讨大棚种植年限对大棚菜田土壤重金属累积、土壤酶活性的影响以及二者的关系,采集不同种植年限(0、5、10、15、20、25、30 a)大棚菜田土壤样品共140份,测定土壤样品中重金属的含量以及土壤酶活性。结果表明:大棚菜田土壤中重金属Zn、Pb、Cu的含量和种植年限极显著相关;重金属Cd、Ni、Mn的含量和种植年限显著相关;重金属Cr的含量和种植年限不相关。大棚菜田土壤中过氧化物酶、多酚氧化酶、淀粉酶活性和种植年限极显著相关,磷酸酶、蔗糖酶活性和种植年限显著相关,过氧化氢酶、脲酶、蛋白酶活性和种植年限相关性不显著。随着种植年限的延长重金属Zn、Cu含量对多酚氧化酶、过氧化物酶活性有抑制作用,其敏感性顺序为:过氧化物酶对Zn敏感性>多酚氧化酶对Zn敏感性>过氧化物酶对Cu敏感性>多酚氧化酶对Cu敏感性。土壤中过氧化物酶、多酚氧化酶可以作为重金属Zn污染的指示酶,过氧化物酶可以作为重金属Cu污染的指示酶。该文为设施污染土壤环境质量评价提供依据。 相似文献
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用平衡吸附法研究了不同浓度的有机酸（乙酸、酒石酸和柠檬酸）对针铁矿和膨润土吸附Cd^2＋、Pb^2＋的影响。结果表明：（1）低浓度（〈0．6mmolL^-1）有机酸促进供试矿物吸附Cd^2＋和Pb^2＋，随着有机酸浓度增加，吸附逐渐被抑制。高浓度（〉1．0mmolL^-1）有机酸对Pb^2＋吸附的抑制作用比对Cd^2＋的强。有机酸浓度的变化对针铁矿Cd^2＋、Pb^2＋吸附率的影响大于对膨润土的。（2）不同Cd^2＋浓度下，有机酸对膨润土Cd^2＋吸附强度的影响大于针铁矿。加入高浓度Cd^2＋（8．0mmolL^-1）时，低浓度有机酸对膨润土吸附Cd^2＋的促进作用比加入低浓度Cd^2＋（0．4mmolL^-1）时明显，高浓度有机酸对膨润土吸附Cd^2＋的抑制作用与低浓度Cd^2＋时相近。（3）低浓度有机酸（〈0．6mmolL^-1）时，酒石酸、柠檬酸对针铁矿吸附Cd^2＋的促进作用大于乙酸，而它们对膨润土吸附Cd^2＋的促进作用相似；在三种有机酸高浓度（〉1．0mmol L^-1）时，针铁矿对Cd^2＋的吸附都趋于稳定，而柠檬酸对膨润土吸附Cd^2＋的抑制作用比乙酸和酒石酸的明显。不同种类有机酸下，矿物对Pb^2＋吸附率大小的变化基本一致。 相似文献
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土壤多酚氧化酶性质研究及意义 总被引：27，自引：2，他引：27
土壤多酚氧化酶是一类以铜、锰为活性中心的氧化还原酶。本文以滨江带生态湿地土壤为材料,系统研究了土壤多酚氧化酶活性随着干湿状态、溶解氧、pH值、温度而变化的情况。结果表明,湿土壤比风干土壤和烘干土壤活性高;氧气充足时活性高于缺氧时活性;多酚氧化酶活性在强酸强碱条件下失活,pH值(9~11)或温度45℃时活性最高。通过对土壤多酚氧化酶性质研究,对一些现象进行了初步解释,如荷花出淤泥而不染成因;为加快土壤多酚氧化酶在环境修复中应用和开发它的广阔应用前景奠定基础。 相似文献
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植物生长调节剂对马铃薯根系理化特性的影响 总被引：5，自引：1，他引：4
在大田栽培条件下,以马铃薯（Solanum tuberosum L.）荷兰212为材料,叶面喷施不同植物生长调节剂,通过比较根体积、根鲜重、根干重的变化,根系活力,根系中多酚氧化酶、淀粉酶和抗坏血酸氧化酶活性的变化,研究了喷施植物生长调节剂对马铃薯根系理化特性的调控效应。结果表明:2-N,N-二乙氨基乙基己酸酯（DTA-6）对根系理化特性的影响较大,它可以有效地调控根系抗坏血酸氧化酶活性的变化规律,在喷药后24 d时,与对照相比,降低了根系的鲜重、体积、干重以及根系活力和淀粉酶活性。SOD模拟物（SODM）可以提高喷药24 d后的根系体积。氯化胆碱（Cc）对根系的形态指标略有影响,对根系活力、淀粉酶以及抗坏血酸氧化酶和多酚氧化酶活性的影响不明显。 相似文献
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以猪粪和秸秆为主要试验材料,添加不同浓度重金属Zn,采取发酵罐处理方法,在好氧高温条件下研究了重金属Zn对猪粪堆肥过程中多酚氧化酶、脱氢酶活性的变化,以及堆腐过程堆体温度、堆料pH值、胡敏酸E4/E6值的变化。结果表明：（1）低量重金属Zn处理（L）较不添加重金属Zn（CK）和添加高量重金属Zn（H）堆料升温快、温度高、高温持续时间长。（2）重金属Zn的加入对堆料的pH值影响不大,不是影响堆肥进程的直接原因。（3）H处理在整个堆肥过程中E4/E6值均高于L和CK,表明高浓度Zn处理抑制腐殖质的缩合和芳构化。（4）L处理的多酚氧化酶活性大多数时间高于H处理的活性,说明低量重金属Zn更好地促进了木质素的降解及其产物的转化。（5）从整个堆肥过程来看,3个不同处理的脱氢酶活性表现出一定的不稳定性,可能是重金属对脱氢酶活性有抑制作用的同时发生＂抗性酶活性＂现象。 相似文献
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莲藕的酶促褐变及其贮藏中褐变的控制 总被引：4，自引：0，他引：4
为有效控制莲藕褐变,试验研究了温度、pH值、底物浓度以及抑制剂对莲藕多酚氧化酶（PPO）的影响。结果表明：莲藕PPO最适pH值为7.5,最适温度为35℃,以邻苯二酚为底物,米氏常数Km为0.0273 mol/L。亚硫酸钠、抗坏血酸（VC）对莲藕PPO活性具有较好抑制作用,乙酸、柠檬酸、植酸、木瓜蛋白酶、半胱氨酸也对莲藕PPO活性具有一定抑制作用。0.2%VC+2%乙酸+0.03%亚硫酸钠抑制剂处理对莲藕褐变的抑制效果最佳。可以通过浸泡亚硫酸钠和抗坏血酸（VC）、调节pH值、低温贮藏等方法来抑制莲藕PPO活性,控制莲藕褐变。 相似文献
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衡阳紫色土丘陵坡地不同植被恢复阶段土壤酶活性特征研究 总被引：5，自引：0，他引：5
本文以典型的衡阳紫色土丘陵坡地不同植被恢复阶段为研究对象,采用空间代替时间序列方法,选用立地条件基本相似的草坡阶段（Grassplot, GT）、 灌草阶段（Frutex and grassplot,FG）、 灌丛阶段（Frutex, FX）和乔灌阶段（Arbor and frutex, AF）,通过调查取样和实验分析,对不同植被恢复阶段的土壤酶、 养分与微生物及其相关性进行了研究。结果表明, 1）随着恢复阶段的演替,脲酶、 多酚氧化酶、 蔗糖酶与过氧化氢酶的活性显著增加,在每个恢复阶段,脲酶、 多酚氧化酶、 蔗糖酶与过氧化氢酶活性随着土层的加深而逐渐减弱,脲酶与多酚氧化酶、 蔗糖酶与过氧化氢酶活性呈显著正相关关系,蔗糖酶与脲酶和多酚氧化酶呈极显著正相关。 2）随着恢复阶段的演替,土壤养分的时空变化与土壤酶活性的变化趋势基本一致,土壤有机碳、 全氮与碱解氮含量呈上升趋势,土壤pH随植被恢复和演替而降低,随土壤深度的增加而上升,与土壤酶活性的变化趋势相反;脲酶与有机碳、 全氮、 碱解氮呈极显著正相关,与pH呈显著负相关,多酚氧化酶与有机碳、 碱解氮呈极显著正相关,与全氮、 速效磷、 速效钾呈显著正相关,与pH呈显著负相关,蔗糖酶活性与有机碳、 全氮、 碱解氮、 速效磷、 速效钾呈显著正相关。 3）不同恢复阶段土壤细菌数量最多,真菌数量和放线菌数量与细菌数量的变化趋势各不相同;细菌平均数量为AF＞FX＞FG＞GT,真菌数量为 FG＞GT＞FX＞AF,放线菌数量为 GT＞FX＞FG＞AF。4）主成分分析揭示脲酶与多酚氧化酶可作为衡阳紫色土丘陵坡地土壤质量评价的指标。研究结果将丰富该地区植物生态学与恢复生态学的内容,为衡阳紫色土丘陵坡地生态系统的恢复与重建提供了重要依据。 相似文献
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通过配合喷施不同浓度亚硒酸钠（Na2SeO3）和醋酸锌[(CH3COO)2Zn·2H2O],研究硒（Se）、锌（Zn）及硒锌交互（Se-Zn）作用对夏茶叶片多酚氧化酶（PPO）活性的影响。结果表明,单施Se（100~200 μg/mL）和单施 Zn（0.4%~0.8%）处理,茶叶叶片PPO活性相对较高;高浓度Se（400 μg/mL）和 Zn（1.2%）处理对PPO活性的提高效应不明显,甚至有一定的抑制作用。同一Se、Zn处理喷施16 d后,茶叶叶片PPO活性相对较高。中浓度Zn（0.4%~0.8%）与中浓度Se（100~200 μg/mL）配合喷施,夏茶叶片多酚氧化酶活性较高;而高浓度的硒锌配施对PPO活性无明显促进效应。同一配施处理,喷施16 d后PPO活性变化趋于稳定,且比喷施8d活性增幅较大。高浓度的Zn与不同浓度的Se配施,PPO活性的时间效应不甚明显,而低中浓度Zn与不同浓度Se配合喷施后,PPO活性的时间效应明显。 相似文献
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Winters AL Minchin FR Michaelson-Yeates TP Lee MR Morris P 《Journal of agricultural and food chemistry》2008,56(8):2817-2824
Polyphenol oxidase (PPO) activity in leaf extracts of wild type (WT) red clover and a mutant line expressing greatly reduced levels of PPO (LP red clover) has been characterized. Both latent and active forms of PPO were present, with the latent being the predominant form. PPO enzyme and substrate (phaselic acid) levels fluctuated over a growing season and were not correlated. Protease activation of latent PPO was demonstrated; however, the rate was too low to have an immediate effect following extraction. A novel, more rapid PPO activation mechanism by the enzyme's own substrate was identified. Rates of protein breakdown and amino acid release were significantly higher in LP red clover extracts compared with WT extracts, with 20 versus 6% breakdown of total protein and 1.9 versus 0.4 mg/g FW of free amino acids released over 24 h, respectively. Inclusion of ascorbic acid increased the extent of protein breakdown. Free phenol content decreased during a 24 h incubation of WT red clover extracts, whereas protein-bound phenol increased and high molecular weight protein species were formed. Inhibition of proteolysis occurred during wilting and ensilage of WT compared with LP forage (1.9 vs 5 and 17 vs 21 g/kg of DM free amino acids for 24 h wilted forage and 90 day silage, respectively). This study shows that whereas constitutive red clover PPO occurs predominantly in the latent form, this fraction can contribute to reducing protein breakdown in crude extracts and during ensilage. 相似文献
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Polyphenol oxidase (E.C. 1.14.18.1) (PPO) extracted from yacon roots (Smallanthus sonchifolius) was partially purified by ammonium sulfate fractionation and separation on Sephadex G-100. The enzyme had a molecular weight of 45 490+/-3500 Da and Km values of 0.23, 1.14, 1.34, and 5.0 mM for the substrates caffeic acid, chlorogenic acid, 4-methylcatechol, and catechol, respectively. When assayed with resorcinol, DL-DOPA, pyrogallol, protocatechuic, p-coumaric, ferulic, and cinnamic acids, catechin, and quercetin, the PPO showed no activity. The optimum pH varied from 5.0 to 6.6, depending on substrate. PPO activity was inhibited by various phenolic and nonphenolic compounds. p-Coumaric and cinnamic acids showed competitive inhibition, with Ki values of 0.017 and 0.011 mM, respectively, using chlorogenic acid as substrate. Heat inactivation from 60 to 90 degrees C showed the enzyme to be relatively stable at 60-70 degrees C, with progressive inactivation when incubated at 80 and 90 degrees C. The Ea (apparent activation energy) for inactivation was 93.69 kJ mol-1. Sucrose, maltose, glucose, fructose, and trehalose at high concentrations appeared to protect yacon PPO against thermal inactivation at 75 and 80 degrees C. 相似文献
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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
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Mosses Okot‐Kotber Allan Liavoga Kwon‐Joong Yong Katherine Bagorogoza 《Cereal Chemistry》2001,78(5):514-520
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. 相似文献
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Sun HJ Wang J Tao XM Shi J Huang MY Chen Z 《Journal of agricultural and food chemistry》2012,60(3):823-829
The purification and partial enzymology characteristics of polyphenol oxidase (PPO) from rape flower were studied. After preliminary treatments, the crude enzyme solution was in turn purified with ammonium sulfate, dialysis, and Sephadex G-75 gel chromatography. The optimal conditions and stability of PPO were examined at different pH values and temperatures. Subsequently, PPO was also characterized by substrate (catechol) concentrations, inhibitors, kinetic parameters, and molecular weight. Results showed that the optimal pH for PPO activity was 5.5 in the presence of catechol and that PPO was relatively stable at pH 3.5-5.5. PPO was moderately stable at temperatures from 60 to 70 °C, whereas it was easily denatured at 80-90 °C. Ethylenediaminetetraacetic acid, sodium chloride, and calcium chloride had little inhibitive effects on PPO, whereas citric acid, sodium sulfite, and ascorbic acid had strongly inhibitive effects. The Michaelis-Menten constant (K(m)) and maximal reaction velocity (V(max)) of PPO were 0.767 mol/L and 0.519 Ab/min/mL of the crude PPO solution, respectively. PPO was finally purified to homogeneity with a purification factor of 4.41-fold and a recovery of 12.41%. Its molecular weight was 60.4 kDa, indicating that the PPO is a dimer. The data obtained in this research may help to prevent the enzymatic browning of rape flower during its storage and processing. 相似文献
16.
小麦籽粒多酚氧化酶（PPO）活性是影响面团褐变的主要原因，检索NCBI网站上注册的小麦PPO基因并对其进行分类及相关变异的研究，对于弄清PPO基因与籽粒PPO活性的关系，开发分子标记选育出含有低PPO活性基因的品种，改良我国面制食品的外观品质有重要作用。本研究通过对NCBI上注册的小麦PPO基因序列的搜索与比对后发现，现有的小麦PPO基因按表达方式可分为两大类（I、II），其中第II大类的PPO基因与小麦籽粒PPO活性密切相关，第II大类第i小类中的PPO基因可能位于小麦2A、2D以外的染色体上，可做为改良面团色泽的侯选基因。通过对具有完整开放阅读框（ORF）的4条PPO基因比对后发现，位于小麦2D染色体长臂上的PPO基因（PPO-2D）存在丰富的等位变异，等位基因间有94个单核苷酸变异（SNP），其中发生在编码区的有80个（cSNP），这些cSNP中有36个影响到基因编码的氨基酸序列，属非同义cSNP。在非同义cSNP处，设计引物（STS-H），对130个已连续测得两年PPO活性的小麦品种进行PCR扩增，结果发现STS-H在大部分低PPO活性品种中没有扩增出目标片段（a），而大部分高PPO活性品种可以扩增出460bp的目标片段（b）。方差分析表明，a、b两种类型品种的PPO活性均值差异达极显著水平（p<0.01），说明非同义cSNP对小麦籽粒PPO活性有重要影响。与STS01（低PPO活性显性标记）比较后发现，STS-H与STS01是一对互补标记。为提高单显性分子标记的使用效率，根据STS01和STS-H引物各自的特点，研究了能同时扩增两对引物的多重PCR反应体系。 相似文献
17.
Carbonaro M Mattera M Nicoli S Bergamo P Cappelloni M 《Journal of agricultural and food chemistry》2002,50(19):5458-5462
Despite the increasing interest in organic products, knowledge about how different levels of fertilization affect nutritionally relevant components is still limited. The concentration of polyphenols and the activity of polyphenoloxidase (PPO), together with the content in ascorbic acid, citric acid, and alpha- and gamma-tocopherol, were assayed in conventional and organic peach (Prunus persica L., cv. Regina bianca) and pear (Pyrus communis L., cv. Williams). 2-Thiobarbituric acid reactive substances and the tocopherolquinone/alpha-tocopherol ratio were used as markers of oxidative damage in fruits. A parallel increase in polyphenol content and PPO activity of organic peach and pear as compared with the corresponding conventional samples was found. Ascorbic and citric acids were higher in organic than conventional peaches, whereas alpha-tocopherol was increased in organic pear. The concentration of oxidation products in organic samples of both fruits was comparable to that of the corresponding conventional ones. These data provide evidence that an improvement in the antioxidant defense system of the plant occurred as a consequence of the organic cultivation practice. This is likely to exert protection against damage of fruit when grown in the absence of pesticides. 相似文献
18.
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. 相似文献
19.
Hou MF Tang XY Zhang WD Liao L Wan HF 《Journal of agricultural and food chemistry》2011,59(21):11456-11460
In this study, polyphenol oxidase (PPO) was extracted from commercial potatoes. Degradation of pentachlorophenol by potato PPO was investigated. The experimental results show that potato PPO is more active in weak acid than in basic condition and that the optimum pH for the reaction is 5.0. The degradation of pentachlorophenol by potato PPO reaches a maximum at 298 K. After reaction for 1 h, the removal of both pentachlorophenol and total organic carbon is >70% with 6.0 units/mL potato PPO at pH 5.0 and 298 K. Pentachlorophenol can be degraded through dechlorination and ring-opening by potato PPO. The work demonstrates that pentachlorophenol can be effectively eliminated by crude potato PPO. 相似文献
20.
This study evaluated the effects of inhibitors on polyphenol oxidase (PPO) activity, the effect of the PPO inhibitor tropolone on noodle darkening, and the correlation of PPO activity with darkening of alkaline noodles. The PPO inhibitors tropolone and salicylhydroxamic acid (each at 1 microM) reduced kernel PPO activity by approximately 50% in three hexaploid wheat cultivars but did not inhibit PPO activity in the two very low PPO cultivars, durum Langdon, and the synthetic hexaploid-derived ID580. Tropolone (100 microg/g flour) inhibited alkaline noodle darkening (deltaL*) by 13-25% in the low PPO wheat cultivar, ID377s, and by 39-54% in the high PPO wheat cultivar, Klasic. Alkaline noodle darkening among 502 wheat samples was correlated with kernel PPO activity (r = 0.64). Results substantiate the hypothesis that PPO plays a major role in darkening of alkaline noodles. However, results also indicate that substantial darkening would occur even at zero PPO activity, as measured in the kernel PPO assay. Therefore, darkening of alkaline noodles is probably due to the cultivar-specific level of PPO activity and the presence of at least one additional darkening mechanism. Further investigation is required to identify the phenolic discoloration agent(s) and to determine the potential roles of non-PPO discoloration mechanisms, both enzymatic and nonenzymatic, in wheat products. 相似文献