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休眠期和萌芽期荸荠球茎中多酚氧化酶(PPO)活性与总酚含量的相关性

Correlation Between Polyphenol Oxidase (PPO) Activity and Total Phenolic Contents in L. Corms During Dormancy and Sprouting Stages.

作者信息

Esmaeili Nardana, Ebrahimzadeh Hassan, Abdi Khosrou

机构信息

Department of Biological Sciences, Texas Tech University, Lubbock, TX 79409, USA.

Department of Plant Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran.

出版信息

Pharmacogn Mag. 2017 Oct;13(Suppl 3):S519-S524. doi: 10.4103/0973-1296.216333. Epub 2017 Oct 11.

Abstract

UNLABELLED

Purification and characterization of polyphenol oxidase (PPO) enzyme and determination of total phenolic contents during dormancy and sprouting stages in corms were performed. PPO enzyme was purified by ammonium sulfate precipitation and ion-exchange chromatography using DEAE-Sephadex A and two isoenzymes were obtained on the SDS-PAGE, which corresponded to molecular weights of 70 and 54 kDa. The values of the enzyme were 4.87 and 2.12 mM for l-DOPA in dormancy and waking stages, respectively. Also, enzyme showed higher values of 0.026 (ΔOD.min) in dormancy compared with the value of 0.019 (ΔOD.min) in waking corms. Results showed an inverse correlation between phenolic contents and PPO activity. Accordingly, it can be concluded that as plant progressed through sprouting stage, in contrast to polyphenol oxidase activity, there was a significant increase in total amount of phenolic compounds, as determined by Folin-Ciocalteu method and water and aqueous ethanol extractions.

SUMMARY

Purification of polyphenol oxidase enzyme using DEAE-Sephadex A in corms.Characterization of polyphenol oxidase enzyme.Comparison of PPO enzyme characteristics in two different physiologic stages of dormancy and sprouting.Determination of phenolic contents.Correlation between phenolic contents and PPO activity during sprouting and dormancy. PPO: Polyphenol Oxidase, DEAE-Sephadex: Diethylaminoethyl Sephadex, SDS-PAGE: Sodium Dodecyl Sulfate- Polyacrylamide Gel Electrophoresis, DOPA: Dihydroxyphenylalanine, PEG: Polyethylene Glycol.

摘要

未标记

对球茎休眠期和发芽期的多酚氧化酶(PPO)进行了纯化、特性鉴定,并测定了总酚含量。采用硫酸铵沉淀和使用DEAE - 葡聚糖A的离子交换色谱法对PPO酶进行纯化,在SDS - PAGE上获得了两种同工酶,其分子量分别为70 kDa和54 kDa。该酶对L - 多巴的Km值在休眠期和苏醒期分别为4.87 mM和2.12 mM。此外,与苏醒球茎中0.019(ΔOD·min)的值相比,该酶在休眠期显示出更高的0.026(ΔOD·min)的Km值。结果表明酚类含量与PPO活性呈负相关。因此,可以得出结论,随着植物进入发芽阶段,与多酚氧化酶活性相反,通过福林 - 西奥尔特法以及水和乙醇水溶液提取法测定,酚类化合物总量显著增加。

总结

使用DEAE - 葡聚糖A对球茎中的多酚氧化酶进行纯化。多酚氧化酶的特性鉴定。比较休眠和发芽两个不同生理阶段的PPO酶特性。酚类含量的测定。发芽和休眠期间酚类含量与PPO活性之间的相关性。PPO:多酚氧化酶,DEAE - 葡聚糖:二乙氨基乙基葡聚糖,SDS - PAGE:十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳,DOPA:二羟基苯丙氨酸,PEG:聚乙二醇。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9d13/5669091/d539bbbcac0e/PM-13-519-g002.jpg

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