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1.
In the present paper, a fully latent polyphenol oxidase (PPO) from desert truffle (Terfezia claveryi Chatin) ascocarps is described for the first time. The enzyme was partially purified by using phase partitioning in Triton X-114 (TX-114). The achieved purification was 2-fold from a crude extract, with a 66% recovery of activity. The interfering lipids were reduced to 13% of the original content. In addition, the purification gave rise to a reduction of phenolic compounds to only 37.5%, thus avoiding the postpurification tanning of the enzyme. Latent PPO was activated by the anionic surfactant sodium dodecyl sulfate (SDS) or by incubation with trypsin. The amount of SDS necessary to obtain a maximum activation was dependent on the nature of the substrate. The use of SDS also permitted the histochemical localization of the latent enzyme within the ascocarp. Terfezia polyphenol oxidase was kinetically characterized using two phenolic substrates (L-DOPA and tert-butylcatechol). The latter substrate presented inhibition at high substrate concentration with a K(si) of 6.3 mM. Different inhibiting agents (kojic and cinnamic acid, mimosine and tropolone) were also studied, tropolone being the most effective.  相似文献   

2.
研究CO2浓度和温度升高对作物各生育期土壤氧化还原酶活性的影响,有助于分析气候变化对土壤养分循环过程的影响。本研究结合人工气候室和盆栽控制实验,模拟3种气候情景（当前环境CO2浓度和温度、仅CO2浓度升高、CO2浓度和温度均升高）和2种水分条件（充分供水和轻度干旱）,研究了谷子（Setaria italica）开花期、开花后10 d、灌浆期和收获期4个生育期土壤氧化还原酶活性对CO2浓度升高和增温的响应。结果表明,CO2浓度由400 μmol mol?1升至700 μmol mol?1显著抑制了土壤过氧化氢酶和多酚氧化酶活性,二者降幅分别为2.86% ~ 7.99%和8.63% ~ 27.00%;而温度由22 ℃增加到26 ℃显著增加了土壤过氧化氢酶和多酚氧化酶活性,二者增幅分别为2.10% ~ 9.83%和10.03% ~ 24.96%;CO2浓度升高和增温的交互作用对两种土壤酶活性的影响在谷子4个生育期均无显著影响。谷子生育期对土壤氧化还原酶活性有显著影响,CO2浓度升高与生育期的交互作用对两种土壤氧化还原酶活性均有显著影响,但增温与生育期交互作用仅对土壤多酚氧化酶活性有显著影响。冗余分析（RDA）结果显示,土壤NH4+和MBN对土壤多酚氧化酶活性的变化有较高的解释度。CO2浓度升高抑制土壤氧化还原酶活性,增温提高土壤氧化还原酶活性,两者在多数谷子生育期表现为拮抗作用;谷子生育期影响土壤氧化还原酶活性对气候变化的响应;土壤有效N含量是影响土壤多酚氧化酶活性的重要因素。  相似文献   

3.
To study the antioxidant activity of quince fruit (pulp, peel, and seed) and jam, methanolic extracts were prepared. Each extract was fractionated into a phenolic fraction and an organic acid fraction and was analyzed by high-performance liquid chromatography (HPLC)/diode array detection and HPLC/UV, respectively. Antiradical activities of the extracts and fractions were evaluated by a microassay using 1,1'-diphenyl-2-picrylhydrazyl. The phenolic fraction always exhibited a stronger antioxidant activity than the whole methanolic extract. Organic acid extracts were always the weakest in terms of antiradical activity, which seems to indicate that the phenolic fraction gives a higher contribution for the antioxidant potential of quince fruit and jam. The evaluation of the antioxidant activity of methanolic extracts showed that peel extract was the one presenting the highest antioxidant capacity. The IC50 values of quince pulp, peel, and jam extracts were correlated with the caffeoylquinic acids total content. Among the phenolic fractions, the seed extract was the one that exhibited the strongest antioxidant activity. The IC50 values of quince pulp, peel, and jam phenolic extracts were strongly correlated with caffeoylquinic acids and phenolics total contents. For organic acid fractions, the peel extract was the one that had the strongest antiradical activity. The IC50 values of quince pulp, peel, and jam organic acid fractions were correlated with the ascorbic acid and citric acid contents.  相似文献   

4.
Latent polyphenol oxidase (LPPO), an enzyme responsible for the browning reaction of sago starches during processing and storage, was investigated. The enzyme was effectively extracted and partially purified from the pith using combinations of nonionic detergents. With Triton X-114 and a temperature-induced phase partitioning method, the enzyme showed a recovery of 70% and purification of 4. 1-fold. Native PAGE analysis of the partially purified LPPO revealed three activity bands when stained with catechol and two bands with pyrogallol. The molecular masses of the enzymes were estimated by SDS-PAGE to be 37, 45, and 53 kDa. The enzyme showed optimum pH values of 4.5 with 4-methylcatechol as a substrate and 7.5 with pyrogallol. The LPPO was highly reactive toward diphenols and triphenols. The activity of the enzyme was greatly enhanced in the presence of trypsin, SDS, ethanol, and linoleic acid.  相似文献   

5.
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   

6.
采用玉米草及海藻寡糖联合修复技术研究了石油污染土壤的修复效果,对修复过程中酶活性变化进行了测定,并采用变性梯度凝胶电泳（DGGE）技术测定了土壤中微生物群落的变化。结果表明,种植玉米草可以有效提高土壤中石油烃的降解,与对照相比石油烃降解率增加了11%;加入不同浓度海藻寡糖进一步增加了石油烃的降解效果,降解率最高达到28.6%。种植植物及加入海藻寡糖可以有效提高多酚氧化酶、脱氢酶及尿酶的活性。PCR-DGGE结果表明植物种植及海藻寡糖的加入增加了土壤中微生物数量,其微生物群落结构与未种植植物及修复前土壤相比发生了较大的变化。  相似文献   

7.
Homogenates were prepared from the leaves of hydroponically cultivated sunflowers (Helianthus annuus, L.) under deficient, normal and toxic B conditions and the polyphenol oxidase activity together with the ortho‐diphenolic and micronutrient content was measured every week from one month to flowering.

B deficiency slightly depressed polyphenol oxidase activity and B toxicity decreased it significantly at all times, the response fitting a line versus the logarithm of the B concentration in solution. We found a positive correlation between polyphenol oxidase activity and the B content in leaves.

B action on enzymatic activity seemed not to be caused by direct interaction with substrates as the infiltration of B into deficient leaves did not restore the normal activity

According to our results we suggest that polyphenol oxidase activity may be a valid functional parameter to indicate the nutritional status of B in sunflower plants, if its specificity for B can be demonstrated.

The ortho‐diphenolic content did not change with the B levels used in our experiments. Neither did we find any correlation between enzymatic activity and the o‐diphenolic content of the leaves.  相似文献   

8.
Pome trees, apple, pear, and quince, are classified into the subfamily Pomoideae, belonging to the Rosaceae family. Their autumnal fruits are consumed worldwide in different forms, that is, fresh or transformed into jams, jelly, juices, etc. Their well-established beneficial properties to human health were found mainly related to their phenolic content. Pulp and peel aqueous acetone extracts obtained from Tunisian fruits at commercial maturity were comparatively evaluated for their phenolic profiles and antioxidant and antimicrobial potentials. The phenolic compounds present in the extracts were identified and quantified using RP-HPLC-DAD and ESI-MS techniques. Significant differences in the chromatographic profiles among these fruits, as well as between pulp and peel extracts of each fruit, were observed. Quince, followed by 'Red Delicious', peel extracts showed the highest phenolic content (160.33 and 110.90 mg/100 g of fresh weight). The stronger inhibitory effect on DPPH radicals corresponded to those obtained from peel materials. A comparative analysis of the antimicrobial potential against a range of microorganism strains was also carried out. Staphylococcus aureus, Pseudomonas aeruginosa, and Bacillus cereus were the most sensitive to the active extracts. Among the examined phenolic extracts, 'Red Delicious' and quince peels showed the highest effects for inhibiting bacteria growth. Minimum inhibitory and bactericide concentrations ranged from 10(2) to 10(4) microg of polyphenol/mL. Red skin apple and quince peels could be of great interest as important antioxidant and antimicrobial polyphenol sources.  相似文献   

9.
A partial characterization of polyphenol oxidase (PPO) activity in Ocimum basilicum L. is described. PPO in O. basilicum L. was extracted and purified through (NH4)2SO4 precipitation, dialysis, and a Sepharose 4B-l-tyrosine-p-aminobenzoic acid affinity column. The samples obtained from (NH4)2SO4 precipitation and dialysis were used for the characterization of PPO. At the end of purification by affinity chromatography, 11.5-fold purification was achived. The purified enzyme exhibited a clear single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular mass of the enzyme was estimated to be approximately 54 kDa. The contents of total phenolic and protein of O. basilicum L. extracts were determined. The total phenolic content of O. basilicum L. was determined spectrophotometrically according to the Folin-Ciocalteu procedure and was found to be 280 mg 100 g(-1) on a fresh weight basis. The protein content was determined according to the Bradford method. The enzyme showed activity to 4-methylcatechol, catechol, and pyrogallol substrates, but not to tyrosine. Therefore, of these three substrates, 4-methylcatecol was the best substrate due to the highest V(max)/K(m) value, followed by pyrogallol and catechol. The optimum pH was at 6, 8, and 9 for 4-methylcatechol, catechol, and pyrogallol, respectively. The enzyme had an optimum temperature of 20, 40, and 50 degrees C for 4-methylcatechol, catechol, and pyrogallol, respectively. It was found that optimum temperature and pH were dependent on the substrates studied. The enzyme activity with increasing temperature and inactivation time for 4-methylcatechol, catechol, and pyrogallol substrates decreased due to heat denaturation of the enzyme.  相似文献   

10.
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   

11.
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   

12.
The effect of different inputs of mineral N on several enzyme activities involved in the C and N cycles was investigated using Oa material of forest floors from four Norway spruce [ Picea abies (L.) Karst.] sites with different C-to-N ratios. The samples from each site were treated with five different concentrations of mineral N (as liquid NH 4NO 3). All samples were incubated aerobically for 15–20 weeks at 15°C and at field capacity. Respiration was measured weekly. At the end of the incubation period, four enzyme activities (endoglucanase, ß-glucosidase, polyphenol oxidase and ß-glucosaminidase) and microbial biomass were determined. Endoglucanase activity was increased and ß-glucosidase activity was decreased by N additions only in Oa material having a wide C-to-N ratio. In N-supplemented samples of low C-to-N ratio, increased polyphenol oxidase activities were often detected as a consequence of N addition. ß-Glucosaminidase activity responded positively to mineral N additions, particularly in Oa samples with low internal N concentration. The results of the present study indicate that the effects of N additions on enzymatic activities of organic matter in late stages of decomposition are related to the C-to-N ratio. Increasing inputs of mineral N to spruce ecosystems may especially affect C-hydrolyzing enzyme activities in soils with wide C-to-N ratio leading to an incomplete degradation of cellulose and thus reduced C availability to micro-organisms.  相似文献   

13.
采用盆栽试验方法,研究了蚯蚓（Pheretima sp.）对土壤-植物系统中菲、芘的去除效果的强化作用及对土壤酶活性的影响。结果表明,在试验浓度范围（0~322 mg.kg-1）内,蚯蚓对供试植物苏丹草（Sorghum vulgare L.）生长促进作用显著。接种蚯蚓的土壤中,苏丹草在菲、芘各浓度下总生物量较未接种蚯蚓土壤分别提高14.41%~25.91%、14.39%~23.9%;蚯蚓活动可促进土壤-植物系统中菲、芘的降解,接种蚯蚓处理中菲、芘各浓度下降解率较未接种蚯蚓处理分别高4.20%~9.76%、3.69%~9.38%。土壤酶活性测定结果也显示蚯蚓活动可增加土壤酶的活性。  相似文献   

14.
长期施肥土壤中酶活性的剖面分布及其动力学特征研究   总被引：13，自引：0，他引：13  
对连续25年的长期培肥试验地中土壤剖面酶活性的分布及土壤酶的催化反应特性进行了研究,结果发现,土壤脲酶、碱性磷酸酶和蔗糖酶活性在土壤剖面中的分布从表层到深层,酶活性依次减小;而多酚氧化酶呈波浪型分布。土壤中脲酶、碱性磷酸酶、蔗糖酶活性与相应土层的有机质含量呈极显著正相关;多酚氧化酶无此相关性。土壤脲酶和碱性磷酸酶活性均随时间的延长而逐渐趋于一个稳定值,且各个时段酶活性均为施肥处理高于不施肥处理,说明施肥能显著提高土壤酶活性。土壤脲酶和碱性磷酸酶酶促反应初速度(V0)均随温度和底物浓度的增加而增加,至一定温度和底物浓度时,V0不再增加;不同培肥措施下V0为:厩肥秸秆化肥对照。  相似文献   

15.
若尔盖湿地土壤酶活性和理化性质与微生物关系的研究   总被引：12，自引：2，他引：10  
本文旨在研究若尔盖湿地土壤退化过程中四种典型土壤(泥炭土、沼泽土、草甸土、风沙土)中脲酶、过氧化氢酶、转化酶、多酚氧化酶活性及其与理化性质、微生物数量的相关性分析。结果表明:随着土壤深度增加,脲酶、转化酶活性逐渐降低,过氧化氢酶、多酚氧化酶活性逐渐升高;随着土壤退化程度的加剧,脲酶和转化酶活性总体呈下降趋势;泥炭土、沼泽土、草甸土中过氧化氢酶活性显著高于风沙土;多酚氧化酶活性随退化程度加剧呈上升趋势,但在沼泽土中活性最低。泥炭土、沼泽土、草甸土中转化酶及风沙土中过氧化氢酶与土壤理化性质、微生物数量的相关性较显著。因此土壤酶活性(脲酶、转化酶)、理化性质(含水量、有机质、全氮、全磷、全钾)、微生物数量能较好地指示当地土壤退化状况。  相似文献   

16.
Twenty-one free amino acids present in several samples of quince fruit (pulp and peel) and quince jam (homemade and industrially manufactured) were analyzed by GC/FID. The analyses showed some differences between quince pulps and peels. Generally, the highest content in total free amino acids and in glycine was found in peels. As a general rule, the three major free amino acids detected in pulps were aspartic acid, asparagine, and hydroxyproline. For quince peels, usually, the three most abundant amino acids were glycine, aspartic acid, and asparagine. Similarly, for quince jams the most important free amino acids were aspartic acid, asparagine, and glycine or hydroxyproline. This study suggests that the free amino acid analysis can be useful for the evaluation of quince jam authenticity. It seems that glycine percentage can be used for the detection of quince peel addition while high alanine content can be related to pear addition.  相似文献   

17.
The influence of procyanidin extract from Japanese quince fruit on the activities of matrix metalloproteinases MMP-2 and MMP-9 secreted to culture medium by human peripheral blood mononuclear cells (PBMC) and by human leukemia HL-60 cells was investigated by gelatin zymography. The extract proved to be an effective inhibitor of the enzymes activities (for MMP-2 and MMP-9 secreted by PBMC IC50 = 16-19 microg extract/mL and 22-25 microg extract/mL, respectively). To identify the most effective components of the extract it was fractionated by means of column chromatography on TSKgel Toyopearl HW-40 (S) bed. The obtained fractions were analyzed by TLC, HPLC, and MALDI-TOF MS. Their antioxidant activity was measured as cation radicals ABTS(.+) scavenging efficiency. The fractions VIII-XIV containing oligomers from trimer to hexamer (and probably higher oligomers) appeared to be the most effective inhibitors of MMP-2 and MMP-9 activity (IC50 value close to 4.6 microg total polyphenols/mL). To the best of our knowledge, it is the first report on gelatinase-inhibitory activity of Japanese quince fruit polyphenol extract. We conclude that polyphenols from Japanese quince can be used in cancer chemoprevention, although further studies are needed to elucidate the mechanisms underlying their biological activities.  相似文献   

18.
熊湖  郑顺林  龚静  黄强  袁继超  何卫 《水土保持学报》2019,33(3):254-259,267
为明确液态有机肥对土壤酚酸导致马铃薯的连作障碍的缓解效应,采用盆栽试验,将阿魏酸与香草酸等量混合后按不同浓度(0,50,100,150 mg/kg)施加于马铃薯基质土壤中,模拟马铃薯连作分泌的有机酸自毒物质,并施加不同浓度梯度液态有机肥(0,225,450,675 kg/hm^2),探讨酚酸胁迫下液态有机肥对马铃薯生长发育以及土壤酶活性的影响。结果表明:单一施加外源酚酸对马铃薯株高、茎粗、叶面积以及干物质量皆有不同程度的抑制作用,随酚酸浓度增加,土壤中脲酶、酸性磷酸酶、蔗糖酶活性相对对照分别降低2.74%~10.95%,11.11%~20.55%,5.29%~12.96%,过氧化氢酶、多酚氧化酶活性升高,FDA水解酶活性表现为低促进高抑制。施加液态有机肥后提高了马铃薯株高、茎粗、叶面积以及干物质量,土壤酶活性均有提高,脲酶、酸性磷酸酶、蔗糖酶、过氧化氢酶、多酚氧化酶、FDA水解酶活性相对对照处理最高分别提高了10.80%,21.40%,18.20%,29.60%,37.69%,12.31%,但是液态有机肥浓度过高降低了其对磷酸酶、脲酶、过氧化氢酶的提升效果,抑制了FDA水解酶活性。因此合理施加液态有机肥可以促进马铃薯生长发育,提高土壤酶活性,增强马铃薯的抗逆性,从而缓解酚酸对马铃薯的胁迫作用。  相似文献   

19.
三种观赏植物对Cd污染土壤酶活性的影响   总被引：1，自引：0，他引：1  
通过盆栽实验,系统研究了含羞草、白雪姬和树马齿苋对Cd污染土壤中土壤酶的修复作用。结果表明,脲酶、过氧化氢酶和磷酸酶的活性随着Cd浓度的上升不断下降,且均与Cd浓度呈显著负相关关系。3种观赏植物生长均能提高Cd污染土壤中脲酶、磷酸酶和过氧化氢酶的活性。白雪姬还能显著提高土壤蔗糖酶的活性。含羞草、白雪姬和树马齿苋对土壤酶的修复效果均表现为:脲酶>磷酸酶>过氧化氢酶>蔗糖酶,且白雪姬对土壤酶的修复效果显著高于其它两种观赏植物,说明白雪姬作为一种常见的观赏植物,在修复重金属Cd污染土壤方面具有很高的应用前景。  相似文献   

20.
接种菌剂对猪粪高温堆肥中酶活性的影响   总被引：10，自引：2，他引：8  
在静态通气条件下,以猪粪为原料,以小麦秸秆作为调节物质,分别用接种复合微生物和常规堆肥两个处理研究了高温堆肥过程中酶活性的变化特征,并对其与堆料E4/E6和电导率（EC）的相互关系进行了探讨。结果表明,接种菌剂堆肥显著地提高了堆料的温度,延长了高温腐解期,有效地提高了猪粪堆肥过程中各种酶的活性和峰值。酶活性的大小因酶种类和堆肥时期的不同而各异;纤维素酶和蔗糖酶在高温期的活性高,在低温期急速下降;脲酶和多酚氧化酶活性峰值出现在堆肥后期低温阶段;脱氢酶活性峰值出现在堆肥中期,过氧化氢酶活性在整个堆肥过程中呈持续下降的趋势。接种菌剂处理和CK处理堆肥期间的EC值均与纤维素酶和脱氢酶活性呈显著的正相关（p＜0.01）;E4/E6与过氧化氢酶活性成显著正相关（p＜0.01）,说明酶活性大小是反映堆肥过程中矿质化过程和腐殖化过程生物化学进程的很好指标。  相似文献   

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