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Polyphenol Oxidase, Peroxidase and PhenylalanineAmmonium Lyase Induced in Postharvest Peach Fruitsby Inoculation with Pichia membranefaciensor Rhizopus stolonifer 被引量:13
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作者 QIN Guo-zheng, TIAN Shi-ping, LIU Hai-bo and XU Yong(Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany , Chinese Academy of Sciences , Beijing 100093 , P. R. China) 《Agricultural Sciences in China》 CAS CSCD 2002年第12期1370-1375,共6页
Rhizopus rot of peach fruits could be significantly suppressed by Pichia membranefaciens. Polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonium-lyase (PAL) activities induced by inoculation with P. mem... Rhizopus rot of peach fruits could be significantly suppressed by Pichia membranefaciens. Polyphenol oxidase (PPO), peroxidase (POD) and phenylalanine ammonium-lyase (PAL) activities induced by inoculation with P. membrane faciens or R. stolonifer were studied in postharvest peach fruits. The activities of PPO and PAL in peaches increased significantly after being inoculated with P. membrane faciens + R. stolonifer by 24 h, the activities maintained at a high level throughout the experiment. Under the condition of infected with R. stolonifer alone, activity of PPO and PAL could also increased, but the levels were lower than those treated with P. membrane faciens+ R. stolonifer. However, fruits inoculaed with P. membrane-faciens + R. stolonifer or R. stolonifer alone did not stimulated POD activity. The results suggest that the activation of these defense enzymes is involved in the action of P. membrane faciens against R. stolonifer. 展开更多
关键词 Peach fruits polyphenol oxidase PERoxidase Phenylalanine ammonium-lyase Pichia membranefaciens Rhizopus stolonifer
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Inhibitory effect of tartary buckwheat seedling extracts and associated flavonoid compounds on the polyphenol oxidase activity in potatoes(Solanum tuberosum L.) 被引量:5
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作者 LI Jun WANG Hui +4 位作者 LU Yang MAO Tang-fen XIONG Jiang HE Sheng-ling LIU Hui 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2019年第9期2173-2182,共10页
To improve the processing quality of potatoes,phosphate buffer extract(PBE),50%ethanol(E50),and aqueous extract(AE)of tartary buckwheat seedlings were evaluated for their ability to inhibit the enzymatic browning of p... To improve the processing quality of potatoes,phosphate buffer extract(PBE),50%ethanol(E50),and aqueous extract(AE)of tartary buckwheat seedlings were evaluated for their ability to inhibit the enzymatic browning of potatoes.The results suggest that all extracts of tartary buckwheat seedlings exert significant inhibitory effects on the polyphenol oxidase(PPO)activity in potatoes.The relative concentrations required for a 50%reduction in the PPO activity(IC50)were 0.21,0.28 and 0.41 mg mL^-1,for PBE,E50 and AE,respectively.The strongest inhibitory activity was observed for PBE,followed by E50 and AE.Four flavone compounds in the PBE of tartary buckwheat seedlings(i.e.,rutin,kaempferol-3-O-rutinoside,quercetin,and kaempferol)were identified by high-performance liquid chromatography.These compounds were subsequently evaluated for their roles in the inhibition of PPO from potatoes using a model system.The results indicated that rutin exhibited the highest inhibition rate on the PPO of potato.A synergistic inhibitory effect was observed by mixing rutin,kaempferol-3-Orutinoside,quercetin,and proteins.The inhibitory patterns of rutin,kaempferol-3-O-rutinoside,and quercetin on the enzyme were noncompetitive and reversible,with inhibitory constants of 0.12,0.31,and 0.40 mg mL^-1,respectively.Flavonoids from tartary buckwheat seedlings may exhibit a common mechanism with phenolic compounds,involving the blockage of the reaction of oxygen with PPO leading to the inhibition of the enzymes involved in browning.Based on these results,extracts of tartary buckwheat seedlings can be used as potent natural inhibitors. 展开更多
关键词 tartary BUCKWHEAT natural inhibitor flavonoid COMPOUNDS polyphenol oxidase POTATO
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Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes 被引量:4
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作者 AMJAD ALI Khan QAYYUM Husain 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 2007年第4期396-402,共7页
In this study an effort has been made to use plant polyphenol oxidases; potato (Solanum tuberosum) and brinjal (Solanum melongena), for the treatment of various important dyes used in textile and other industries.... In this study an effort has been made to use plant polyphenol oxidases; potato (Solanum tuberosum) and brinjal (Solanum melongena), for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation. 展开更多
关键词 DECOLORIZATION textile dyes polyphenol oxidases POTATO BRINJAL wastewater treatment removal of dyes
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Effects of Terpinen-4-ol on Four Metabolic Enzymes and Polyphenol Oxidase (PPO) in Mythimna separta Walker 被引量:4
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作者 MA Zhi-qing HAN Xiu-ling FENG Jun-tao LI Guang-ze ZHANG Xing 《Agricultural Sciences in China》 CAS CSCD 2008年第6期726-730,共5页
To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical app... To study insecticidal mechanism of terpinen-4-ol, a main insecticidal composition in the essential oil of Sabina vulgaris, the 5th instar larvae of Mythimna separta, were investigated with terpinen-4-ol by topical application. The activities of phosphatase, glutathione S-transferase (GSTs), cytochrome P450 (P450), and polyphenol oxidase (PPO) of tested insects were determined in all poisoning stages, including exciting stage, convulsing stage, paralysis stage, and recover stage. The result showed that the activities of both acid phosphatase (ACP) and alkaline phosphatase (AKP) in treated insects were induced by terpinen-4-ol, but ACP was inhibited in paralysis stage. The activities of GSTs were inhibited in exciting stage, convulsing stage, and paralysis stage, but gradually recovered in recover stage. O-demethylase activity of cytochrome P450 was inhibited by terpinen-4-ol, and the inhibition rate in all poisoning stages were 26.27, 46.03, 80.24, and 90.22%, respectively. PPO activities were strongly inhibited by terpinen-4-ol both in vitro and in vivo. In conclusion, the activities of P450, GSTs, and PPO could have relation with toxicity of terpinen-4-ol against larvae of the Mythimna separta, but recover stage of the poisoning insects might be related to GSTs induced. As a new insecticide or synergist, terpinen- 4-ol has a potential value in field of insecticide resistance management. 展开更多
关键词 TERPINEN-4-OL Mythimna separate polyphenol oxidase (PPO) metabolic enzymes insecticidal mechanism
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Study on Enzymatic Kinetics of Nephelium lappaceum Polyphenol Oxidase 被引量:2
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作者 Lunpei LIU Wenhua ZHANG 《Agricultural Biotechnology》 CAS 2012年第4期39-42,共4页
[Objective] This study aimed to investigate the enzymatic kinetics of polyphenol oxidase in peel of Nephelium lappaceum to explore the environmental factors affecting its catalytic activity. [ Method ] With N. lappace... [Objective] This study aimed to investigate the enzymatic kinetics of polyphenol oxidase in peel of Nephelium lappaceum to explore the environmental factors affecting its catalytic activity. [ Method ] With N. lappaceum peel as the experimental material and catechol as substrate, effects of temperature, pH, four inhibitors (EDTA, Vc, sodium bisulfite and citric acid) and substrate concentration on the activity of polyphenol oxidase were investigated. [ Result] The optimal temperature for polyphenol oxidase in N. lappaceum peel was 40 %, and the optimal pH was 6.8. EDTA, Vc, sodium bisulfite and citric acid all showed ideal in- hibitory effects on the activity of polyphenol oxidase; specifically, EDTA had the strongest inhibitory effect. [ Conclusion] At low temperature, EDTA, Vc, sodi- um bisulflte and citric acid with certain concentrations can inhibit the catalytic activity of polyphenol oxidase in peel of N. lappaceum, extend the storage and trans- portation time of N. lappaceum fruit, and inhibit the browning. This study provides theoretical basis for the development and utilization of polyphenol oxidase in peel of N. lappaceum. 展开更多
关键词 Nephelium lappaceum polyphenol oxidase Enzymatic kinetics
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Study on Polyphenol Oxidase Activity and Total Phenol Content of Docynia indica 被引量:1
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作者 Xiaobo QIN Xinyi XU +4 位作者 Bei NIU Xiaodong SHI Bingbing ZHANG Lijuan FAN Dongmei HU 《Medicinal Plant》 CAS 2019年第4期73-75,83,共4页
[Objectives] To study the polyphenol oxidase (PPO) activity and total phenol content of Docynia indica .[Methods] The tender branches and petioles of the medicinal and edible plant D. indica were used as experimental ... [Objectives] To study the polyphenol oxidase (PPO) activity and total phenol content of Docynia indica .[Methods] The tender branches and petioles of the medicinal and edible plant D. indica were used as experimental materials, and the annual changes of total phenol content and polyphenol oxidase (PPO) activity were analyzed.[Results] pH 7.0 was the optimum value of polyphenol oxidase activity in D. indica plants;the best substrate was catechol with the concentration of 0.15 mol/L;the optimum temperature was 25 ℃.[Conclusions] The total phenol content of D. indica plants was the lowest in April, May and September of each year. The polyphenol oxidase activity of D. indica plants at flowering and fruiting stage was significantly higher than that at pre-flowering stage. 展开更多
关键词 Docynia INDICA polyphenol oxidase (PPO) activity TOTAL PHENOL content
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Purification and Partial Characterization of Polyphenol Oxidase from Sapodilla Plum (<i>Achras sapota</i>) 被引量:1
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作者 Jorge Tamayo Cortez Carlos Hernán Herrera Méndez +2 位作者 Enrique Sauri Duch María de Lourdes Vargas y Vargas Sara Solís Pereira 《Food and Nutrition Sciences》 2013年第7期727-734,共8页
The browning of fruits can be considered as an enzymatic oxidation which is believed to be one of the main causes of quality loss during post-harvest handling. The enzymes responsible for this are the oxidoreductases;... The browning of fruits can be considered as an enzymatic oxidation which is believed to be one of the main causes of quality loss during post-harvest handling. The enzymes responsible for this are the oxidoreductases;the polyphenol oxidase (PPO) (monophenol, o-diphenol, oxygen oxidoreductase;EC 1.14.18.1) belongs to this group. This enzyme, which is found in the sapodilla plum (Achras sapota), was purified using a phenylsepharose and a SephacrylS-200 columns. The molecular weight of the purified enzyme was estimated to be about 66 kDa by gel filtration and 29 kDa by SDS-PAGE. A single protein band was found using the latter system (SDS-PAGE), which shows that the PPO of the pulp of the sapodilla plum may be composed of two protein subunits with similar molecular weight. The optimum pH was 7.0 and the optimum temperature 60℃. The most effective inhibitors tested were ascorbic acid, sodium metabisulfite and acetic acid. 展开更多
关键词 Enzymatic System Oxidation polyphenol oxidase BROWNING Process Nutritional Value
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Polyphenol Oxidase Inactivation by Microwave Oven and Its Effect on Phenolic Profile of Loquat (<i>Eriobotrya japonica</i>) Fruit 被引量:1
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作者 Yanet Chávez-Reyes Lidia Dorantes-Alvarez +2 位作者 Daniel Arrieta-Baez Obed Osorio-Esquivel Alicia Ortiz-Moreno 《Food and Nutrition Sciences》 2013年第9期87-94,共8页
The objective of this research was investigated the effect of polyphenol oxidase microwave treatment on phenolic composition, antioxidant activity and microstructure of loquat fruit. Phenolic profile of methanolic ext... The objective of this research was investigated the effect of polyphenol oxidase microwave treatment on phenolic composition, antioxidant activity and microstructure of loquat fruit. Phenolic profile of methanolic extracts prepared from fresh, and microwave-treated samples were analyzed. Antioxidant activity was also evaluated by 2,2’-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS?+) and 1,1-diphenyl-2-picrylhydrazyl (DPPH+) methods. In addition, polyphenol oxidase inactivation was carried out using a response surface methodology to establish the optimal conditions of treatment. The phenolic content of fresh mesocarp was 311 ± 0.60 mg gallic acid equivalents (GAE)/100g dry weight (DW) and that of microwave-treated mesocarp was 1230 ± 0.36 mg GAE/100g DW. Total phenolic content of water/ methanol extract significantly increases after microwave treatment rather than methanolic extract of fresh loquat. Five glycoside phenolics were identified by HPLC-DAD-MS as 3-caffeoylquinic acid, 3-p-coumaroylquinic acid, 5-caffeoylquinic acid and quercetin-3-O-sambubioside. Methanolic extract of microwave-treated mesocarp showed higher antioxidant activity than that of fresh mesocarp. Thus, polyphenol oxidase inactivation by microwave energy preserved the integrity of phenolic compounds as well as antioxidant activity in mesocarp extracts prepared from loquat fruit. It was also noted that phenolics were more abundant in the microwaved samples than in the fresh samples. 展开更多
关键词 LOQUAT PHENOLIC Compounds polyphenol oxidase Microwave ERIOBOTRYA japonica
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Polyphenol Oxidase Activity and Inhibition in White Yam (Dioscorea Rotundata. Var. Laasirin) Chips as African Fries for Human Consumption 被引量:1
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作者 Oluwatoyin Oluwole Olajumoke Odediran +6 位作者 Gbolahan Alagbe Ijeoma Eboagwu Abimbola Jegede Kayode Ogundeji Abimbola Olokoshe Godfrey Asieba Joy Onyibe 《Journal of Food Science and Engineering》 2016年第1期43-50,共8页
Polyphenol oxidase, a bi-functional enzyme, has been implicated in enzymatic browning of yam and other tubers in a negative way. The objective of this present study was to examine the activity of polyphenol oxidase in... Polyphenol oxidase, a bi-functional enzyme, has been implicated in enzymatic browning of yam and other tubers in a negative way. The objective of this present study was to examine the activity of polyphenol oxidase in Dioscorea rotundata. Var. laasirin and the efficiency of heat and chemical treatments in inhibiting this enzyme. Crude Polyphenol Oxidase (PPO) of Dioscorea rotundata.Var. Laasirin was isolated and the kinetics studied using the lineweaver-burk plot. The activity of the enzyme was evaluated using spectrophotomeric method. Yam PPO catalyzes oxidation of various substrates with catechol being the most readily oxidized substrate. The Michaelis-Menten constant (Km) and maximum reaction velocity (Vmax) for yam PPO were 0.00037 and 0.3125 respectively. Inhibition data showed that the enzyme had least activity (71.70) when blanched at 95℃ for 7 mins with chemical treatment involving a combination of 0.5% Sodium metabisulphite (Food grade) and 0.5% Ascorbic acid (Food grade). The activity was highest (83.02) when it was blanched at 95 ℃ for 7 rains. This study has shown that it is possible to inhibit polyphenol oxidase activity in white yam using the chemical pretreatments and processing conditions described in this study for possible adoption in the production of packaged frozen yam chips by food industries. 展开更多
关键词 YAM polyphenol oxidase dioscorea rotundata blanch SULPHITE chips.
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Polyphenol Oxidase Activity and Colour Changes of ‘Starking’ Apple Cubes Coated with Alginate and Dehydrated with Air
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作者 Susana Daniela Sousa Fernandes Catarina Araújo da Silva Ribeiro +2 位作者 Maria Filomena de Jesus Raposo Rui Manuel Santos Costa de Morais Alcina Maria Miranda Bernardo de Morais 《Food and Nutrition Sciences》 2011年第5期451-457,共7页
The objective was to study the effect of alginate coating on polyphenol oxidase (PPO) activity and colour of ‘Starking’ apple cubes during dehydration with hot air. Apple cubes were dehydrated at 20oC, 35oC, or 40oC... The objective was to study the effect of alginate coating on polyphenol oxidase (PPO) activity and colour of ‘Starking’ apple cubes during dehydration with hot air. Apple cubes were dehydrated at 20oC, 35oC, or 40oC, with a parallel airflow. Analysis of PPO activity, colour (L*, a*, b*) and dry matter were performed along the dehydration process at each temperature. All samples presented a peak in relative PPO activity in the beginning of the drying. Exponential models fitted well the experimental data after the peak. Cubes without coating presented lower PPO activity, suggesting lower browning, than coated samples throughout the dehydration process, for all temperatures. Better results for coated samples were obtained with a perpendicular airflow drying at 40oC, after dipping the whole apple in water at 60oC for 10 min. In order to prevent coated samples from browning, drying by perpendicular airflow preceded by a thermal treatment of the whole apple is required. 展开更多
关键词 ALGINATE Coating APPLE CUBES Colour Hot AIR Drying polyphenol oxidase
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Immobilization and Properties of Polyphenol Oxidase
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作者 Zheng LI Pan WANG +4 位作者 Yanxi SHEN Mingfei WEI Yimin HE Aoyu JI Jianfeng ZHAN 《Agricultural Biotechnology》 CAS 2021年第6期79-83,共5页
[Objectives]This study was conducted to investigate the effects of embedding conditions on activity and catalytic properties of immobilized polyphenol oxidase.[Methods]Polyphenol oxidase was immobilized in a polymer m... [Objectives]This study was conducted to investigate the effects of embedding conditions on activity and catalytic properties of immobilized polyphenol oxidase.[Methods]Polyphenol oxidase was immobilized in a polymer material by the embedding method,and the optimal immobilization conditions were obtained by single factor tests:CaCl_(2) concentration 2.0%,sodium alginate concentration 2.0%,immobilization time 2 h and mass ratio of enzyme to carrier 10 mg/100 g,under which the immobilized enzyme activity was 93.33 U/g.Under the above conditions,the properties of polyphenol oxidase immobilized by sodium alginate(A-PPO)and free polyphenol oxidase were studied.[Results]The thermostability of A-PPO was better than that of the free enzyme,but the pH stability of A-PPO was inhibited.The Michaelis constant K_(m) values of free polyphenol oxidase and A-PPO were 0.37 and 0.48 mmol/L,respectively,and the maximum reaction rate V_(max) values were 0.38 and 0.51 mmol/(L·g),respectively.[Conclusions]This study provides a theoretical basis for the study of the properties of polyphenol oxidase. 展开更多
关键词 polyphenol oxidase Immobilized enzyme Sodium alginate Enzymatic properties Kinetic parameter
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<i>In Silico</i>and <i>in Vitro</i>Approach for the Understanding of the Xanthine Oxidase Inhibitory Activity of Uruguayan Tannat Grape Pomace and Propolis Poliphenols
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作者 Elena Alvareda Federico Iribarne +5 位作者 Victoria Espinosa Pablo Miranda Daniela Santi Sara Aguilera S. Bustos Margot Paulino Zunini 《Journal of Biophysical Chemistry》 2019年第1期1-14,共14页
The use of food additives with xanthine oxidase (XO) inhibitory activity offers an alternative approach to hyperuricemic and gout disease treatment, and provides an example of antioxidant nutraceutics. The in vitro an... The use of food additives with xanthine oxidase (XO) inhibitory activity offers an alternative approach to hyperuricemic and gout disease treatment, and provides an example of antioxidant nutraceutics. The in vitro and in silico XO inhibitory activity of polyphenols from Uruguayan Tannat grape pomaces and propolis extracts was evaluated as well as the scavenging capacity of said compounds. When comparing propolis and grape pomace samples, the in vitro studies demonstrated that polyphenols extracted from propolis are more active as free radical scavengers than those from Tannat grape pomace. Both natural products effectively inhibited XO but the capacity of phenols present in GP is higher than the one present in P. The high content of anthocyanins in GP, absent in P, could account for this observation. In silico assays allowed us to determine relevant ligand-receptor interactions between polyphenols, from a database built with previously reported polyphenols from both natural products, and the active site of XO. The in silico results showed that compound (E)-isoprenylcaffeate from propolis was the best potential XO inhibitor displaying hydrophobic aromatic interaction between the conjugated ring of the caffeate moiety and polar interactions between hydroxyl groups from caffeate with the active site polar residues. Among grape pomaces, the Cyanidin-3-O-(6-(E)-p-coumaroyl)-glucoside was the best XO inhibitor;its moiety oxychromenyl being relevant to the docking stabilization. All these results lead us to propose Uruguayan propolis and Tannat grape pomace extracts as food additives as well as phytopharmaceuticals to decrease the uric acid levels in gout disease and to act against oxidative stress. 展开更多
关键词 PROPOLIS Tannat Grape POMACE XANTHINE oxidase Inhibition polyphenols Functional Foods
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采前氨基乙氧基乙烯甘氨酸处理对‘黄冠’梨长期冷藏后果实品质和果心褐变的影响
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作者 何近刚 冯云霄 +1 位作者 程玉豆 关军锋 《食品科学》 EI CAS CSCD 北大核心 2024年第3期159-166,共8页
为延长‘黄冠’梨贮藏期,减轻长期贮藏后货架期间果实品质劣变,采用不同浓度氨基乙氧基乙烯甘氨酸(aminoethoxyvinylglycine,AVG)在采前喷施‘黄冠’梨果实,在冷藏180 d及货架7 d((180+7)d)后检测相关指标。结果显示,采前喷施AVG可延缓... 为延长‘黄冠’梨贮藏期,减轻长期贮藏后货架期间果实品质劣变,采用不同浓度氨基乙氧基乙烯甘氨酸(aminoethoxyvinylglycine,AVG)在采前喷施‘黄冠’梨果实,在冷藏180 d及货架7 d((180+7)d)后检测相关指标。结果显示,采前喷施AVG可延缓‘黄冠’梨果实硬度下降,维持可溶性固形物含量以及可滴定酸含量,有效抑制果面褐斑和果心褐变的发生,其中以200 mg/L AVG效果最好。‘黄冠’梨果实在长期冷藏时果心发生褐变,同时熊果苷、绿原酸以及总酚含量增加,多酚氧化酶活性升高;PbPAL1、PbPAL2、PbPPO1、PbPPO5、PbLOX1以及PbLOX5在冷藏期间表达量升高,货架期表达量随之下降。采前200 mg/L AVG处理‘黄冠’梨,可显著促进熊果苷、绿原酸以及总酚在整个贮藏过程中含量的增加,抑制多酚氧化酶活性升高,抑制PbPAL1、PbPAL2、PbPPO1、PbPPO5以及PbLOX5的表达,进而有效减少果心褐变的发生。 展开更多
关键词 氨基乙氧基乙烯甘氨酸 果心褐变 酚类 多酚氧化酶
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大肠杆菌多酚氧化酶的分子克隆及异源高效表达
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作者 邓卉 余丹 +6 位作者 邹成义 范景胜 李斌 屈东 倪青松 郑钰嘉 陈瑾 《中国饲料》 北大核心 2024年第5期26-31,共6页
菜籽粕、棉籽粕等非粮饲料资源存在的酚类化合物芥子碱、棉酚等抗营养因子,严重影响了其饲喂价值。本课题组筛选出的一株大肠杆菌属芥子碱降解菌SDB2已被证实能通过分泌多酚氧化酶来降解芥子碱和棉酚,本试验旨在运用基因工程技术克隆SDB... 菜籽粕、棉籽粕等非粮饲料资源存在的酚类化合物芥子碱、棉酚等抗营养因子,严重影响了其饲喂价值。本课题组筛选出的一株大肠杆菌属芥子碱降解菌SDB2已被证实能通过分泌多酚氧化酶来降解芥子碱和棉酚,本试验旨在运用基因工程技术克隆SDB2多酚氧化酶基因,实现其高效表达,为SDB2多酚氧化酶在饲料工业上的规模化应用奠定基础。从大肠杆菌SDB2中克隆出多酚氧化酶基因,将其与质粒pET28a连接后转化至大肠杆菌BL21感受态细胞中培养,通过PCR鉴定及质粒酶切验证方法构建重组菌株,同时对重组克隆基因进行生物信息学分析。采用“2×2”交叉试验设计,以温度和诱导剂IPTG浓度两个影响因素诱导SDB2多酚氧化酶基因在重组菌株中高效表达,经SDS-PAGE和WB双重验证后,确立最佳诱导条件,最终纯化出目标酶蛋白。结果表明:(1)成功克隆SDB2多酚氧化酶基因,构建出异源表达重组菌株。(2)克隆出的SDB2重组多酚氧化酶基因含目标核苷酸741个,总编码氨基酸263个(含标签氨基酸20个),氨基酸组成的蛋白相对分子质量为28499.0,理论等电点为6.94,据此预测出SDB2重组多酚氧化酶三维结构模型。(3)确立SDB2多酚氧化酶异源高效表达的最佳诱导条件为温度37℃,IPTG浓度1 mmol,该条件下表达出大量可溶性酶蛋白,有利于工业化生产。本试验条件下,大肠杆菌SDB2多酚氧化酶成功实现了分子克隆及异源高效表达,为我国非粮饲料资源实现高效利用提供了技术参考。 展开更多
关键词 大肠杆菌 多酚氧化酶 克隆 高效表达
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蜂蜜对苹果酶促褐变的抑制及有效成分分析 被引量:1
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作者 黄欣莹 姚卫蓉 《食品与生物技术学报》 CAS CSCD 北大核心 2024年第2期55-62,共8页
作者以防止新鲜苹果切口的褐变为目标,比对不同食品配料组成的复配成分,发现蜂蜜作为涂膜材料可以有效延缓鲜切苹果片的褐变。将鲜切苹果片浸泡在质量分数0.1 g/g的椴树蜂蜜水溶液中,8 d后其仍能维持新鲜的色泽;探究了蜂蜜对苹果多酚氧... 作者以防止新鲜苹果切口的褐变为目标,比对不同食品配料组成的复配成分,发现蜂蜜作为涂膜材料可以有效延缓鲜切苹果片的褐变。将鲜切苹果片浸泡在质量分数0.1 g/g的椴树蜂蜜水溶液中,8 d后其仍能维持新鲜的色泽;探究了蜂蜜对苹果多酚氧化酶(polyphenol oxidase,PPO)抑制的动力学,发现蜂蜜对苹果PPO起非竞争性抑制作用,酶活抑制能力来自蜂蜜的抗氧化、清除自由基(相关系数r>0.6)能力;进一步分析蜂蜜成分与酶活抑制率的关系,发现蜂蜜中蛋白质与多酚(r值分别为0.79、0.63)是蜂蜜发挥抗褐变作用的重要成分。 展开更多
关键词 蜂蜜 鲜切苹果片 多酚氧化酶 抗褐变
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分子动力学模拟分析超高压对固液两态多酚氧化酶稳定性的影响
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作者 李镜浩 梁展鸿 +6 位作者 肖更生 徐玉娟 余元善 吴继军 彭健 李璐 程丽娜 《食品科学》 EI CAS CSCD 北大核心 2024年第16期33-44,共12页
以固态晶体(crystal,C-)、液态(liquid,L-)多酚氧化酶(polyphenol oxidase,PPO)为对象,采用分子动力学模拟的方法,分析其在常温(298.15 K)常压/高压(0.1~400.0 MPa)条件下分子构象的变化。结果表明:固液两态PPO的空间结构均随压力(0.1~4... 以固态晶体(crystal,C-)、液态(liquid,L-)多酚氧化酶(polyphenol oxidase,PPO)为对象,采用分子动力学模拟的方法,分析其在常温(298.15 K)常压/高压(0.1~400.0 MPa)条件下分子构象的变化。结果表明:固液两态PPO的空间结构均随压力(0.1~400.0 MPa)增大而变的不稳定;与L-PPO相比,C-PPO对压力更敏感,表现出更高的残基波动,溶剂可及表面积和体积减小,蛋白结构更致密;α-螺旋向无规卷曲过渡,导致氢键数量不稳定,螺旋弹性也降低,整体构象差异明显;活性位点Cu^(2+)位置疏远,残基之间距离发生改变、运动出现随机化,干扰底物催化结合。因此,物理状态和压力改变PPO空间结构、残基运动模式和底物结合范围,影响酶稳定性。因此,PPO状态和压力因素对酶变性程度可排序为:固态>液态,物理状态>压力水平。 展开更多
关键词 多酚氧化酶 分子动力学模拟 物理状态 超高压 稳定性
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灰霉病早期胁迫下花椰菜多酚氧化酶活性的高光谱研究
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作者 王凯 薛建新 +1 位作者 李尧迪 张明月 《光谱学与光谱分析》 SCIE EI CAS CSCD 北大核心 2024年第2期534-541,共8页
利用高光谱技术实现灰霉病早期胁迫下花椰菜多酚氧化酶(PPO)活性的快速无损检测。为了使预测效果更好,在900~1700 nm光谱范围内采集253个健康花椰菜样本及257个染病花椰菜样本的光谱信息,并使用分光光度计法对样本中多酚氧化酶活性进行... 利用高光谱技术实现灰霉病早期胁迫下花椰菜多酚氧化酶(PPO)活性的快速无损检测。为了使预测效果更好,在900~1700 nm光谱范围内采集253个健康花椰菜样本及257个染病花椰菜样本的光谱信息,并使用分光光度计法对样本中多酚氧化酶活性进行测定。对健康及染病花椰菜样本PPO活性均值进行分析,发现健康花椰菜PPO活性均值(10.257 U·g^(-1))小于染病花椰菜PPO活性均值(12.324 U·g^(-1))。利用光谱-理化值共生距离(SPXY)算法对样本进行校正集(健康样本193个,染病样本197个)和预测集(健康样本60个,染病样本60个)的划分,对划分后的样本集进行六种单一预处理(卷积平滑算法SG、去趋势算法DT、中值滤波MF、归一化处理NOR、标准正态变量变换SNV、基线校正Baseline)。利用相关系数(R)和均方根误差(RMSE)作为模型评价指标,发现预处理能够有效提高模型的精度和稳定性,其中健康样本经NOR预处理后的预测集建模效果最好;染病样本经DT预处理后的预测集建模效果最好。采用连续投影算法(SPA)与回归系数法(RC)提取特征波长,建立PPO活性值的偏最小二乘回归(PLSR)、最小二乘支持向量机(LS-SVM)及BP神经网络三种预测模型,探究不同特征波长提取方法对模型精度的影响,比较不同建模方式对花椰菜PPO活性预测的准确度。结果说明提取特征波长可以优化光谱信息,SPA和RC对两种样本提取的波长个数分别为9,12,7和11,其中SPA优化后的光谱数据对健康样本PPO活性预测效果较好,RC优化后的光谱数据对染病样本PPO活性预测效果较好。对比分析模型效果,发现LS-SVM模型对两种样本和其对应的酶活性产生了很好的拟合效果。最终发现,SPA-LS-SVM模型对于健康花椰菜PPO活性预测效果较好,其预测相关系数(Rp)为0.832,预测均方根误差(RMSEP)为1.676;RC-LS-SVM模型对于染病花椰菜PPO活性预测效果较好,其Rp为0.848,RMSEP为1.156。研究结果表明高光谱技术可以实现灰霉病早期胁迫下花椰菜多酚氧化酶活性的测定,为快速检测花椰菜多酚氧化酶活性及便携式仪器开发提供了科学依据。 展开更多
关键词 花椰菜 灰霉病 多酚氧化酶 高光谱 无损检测
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月柿果片不同部位酚类组成及褐变酶活力分布特征
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作者 吴婷婷 段振华 +1 位作者 田玉红 王春婷 《食品研究与开发》 CAS 2024年第10期1-6,43,共7页
为研究月柿果片不同部位酚类组成与其在微波干燥过程中色泽变化不均匀的关系,将月柿果片由顶到底分为A、B、C 3个区域,由外到内分成1、2两个区域,从酶促褐变视角探究新鲜月柿果片不同部位水分、总酚、可溶性单宁、总黄酮含量及多酚氧化... 为研究月柿果片不同部位酚类组成与其在微波干燥过程中色泽变化不均匀的关系,将月柿果片由顶到底分为A、B、C 3个区域,由外到内分成1、2两个区域,从酶促褐变视角探究新鲜月柿果片不同部位水分、总酚、可溶性单宁、总黄酮含量及多酚氧化酶(polyphenol oxidase,PPO)、过氧化物酶(peroxidase enzyme,POD)活力的分布特征。结果表明,在纵向方向上,水分含量由顶到底依次减少,POD活力则呈现相反的趋势;总酚和总黄酮含量分布为A区>C区>B区;可溶性单宁含量呈现A1区<B1区<C1区、B2区<C2区<A2区的变化趋势;PPO活力呈现B区>C区>A区的分布特征。在横向方向上,水分含量和PPO、POD活力分布表现为1区<2区;总酚、可溶性单宁和总黄酮含量呈现A1区<A2区、B1区>B2区、C1区>C2区的变化趋势。此外,主成分分析表明月柿果片不同部位活性成分组成有明显差异。 展开更多
关键词 月柿果片 不同部位 酚类物质 多酚氧化酶 过氧化物酶
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马铃薯抗褐变品种的鉴定与筛选
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作者 商靖雯 石瑛 《中国马铃薯》 2024年第1期10-20,共11页
褐变是马铃薯贮藏和加工过程中常见现象,影响其营养、感官和食味品质,降低其商品价值。以24个马铃薯品种为试验材料,通过测定多酚氧化酶(Polyphenol oxidase,PPO)活性、褐变强度、维生素C含量和褐化指数4个相关指标,对不同马铃薯品种抗... 褐变是马铃薯贮藏和加工过程中常见现象,影响其营养、感官和食味品质,降低其商品价值。以24个马铃薯品种为试验材料,通过测定多酚氧化酶(Polyphenol oxidase,PPO)活性、褐变强度、维生素C含量和褐化指数4个相关指标,对不同马铃薯品种抗褐变能力进行综合鉴定和评价。不同马铃薯品种PPO活性、褐变强度、维生素C含量、褐化指数均存在明显差异。选择PPO活性在15 U以内的11个品种为预选褐变材料;选择维生素C含量在8 mg/100 g以上的7个品种为预选褐变材料;选择初始褐变强度小于0.2,变化值在0.5以内的7个品种为预选褐变材料;选择24 h总褐化指数在Ⅱ级以内,且开始发生褐变时间大于1 h的11个品种为预选褐变材料。对4个褐变相关指标进行相关性和聚类分析,将测试的24个马铃薯品种聚为3类。类型Ⅰ为易褐变类型,类型Ⅱ是中间水平类型,类型Ⅲ是抗褐变类型。筛选出6个抗褐变马铃薯品种,即‘东农314’‘东农324’‘东农325’‘东农328’‘延薯13号’和‘中薯5号’。研究可为马铃薯种质资源创新和抗褐变育种提供基础材料。 展开更多
关键词 马铃薯 抗褐变 多酚氧化酶 褐化指数 褐变强度
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牛油果酶促褐变复合抑制措施研究
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作者 吴莉莎 刘金晶 +2 位作者 李卓霞 牟黠 方莹 《中南民族大学学报(自然科学版)》 CAS 2024年第5期608-613,共6页
以牛油果为研究对象,通过对其果肉中多酚氧化酶(polyphenol oxidase,PPO)的活性进行探究,预测不同理化因素(pH值、超声、化学抑制剂等)对牛油果酶促褐变的影响.首先利用单因素实验分别研究不同pH值、超声时长、化学抑制剂浓度对牛油果果... 以牛油果为研究对象,通过对其果肉中多酚氧化酶(polyphenol oxidase,PPO)的活性进行探究,预测不同理化因素(pH值、超声、化学抑制剂等)对牛油果酶促褐变的影响.首先利用单因素实验分别研究不同pH值、超声时长、化学抑制剂浓度对牛油果果肉PPO活性的影响,进一步通过正交试验确定最佳复合酶促褐变抑制措施.结果表明,最佳复合抑制方案为:pH7.0,超声处理时长40 min,抗坏血酸添加浓度0.1%,L-半胱氨酸添加浓度0.0375%,7 d内储存的酶促褐变抑制率为93.9%.研究为牛油果及其副产品的储存与加工提供了有效的保鲜方法,有利于提高牛油果的食用品质和商品价值. 展开更多
关键词 牛油果 酶促褐变 多酚氧化酶 正交试验 复合抑制措施
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