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象腿芋多酚氧化酶的生化特性及热失活研究

Biochemical characterization and thermal inactivation of polyphenol oxidase from elephant foot yam ().

作者信息

Singh Anuradha, Wadhwa Neeraj

机构信息

Department of Biotechnology, Jaypee Institute of Information Technology, A-10,sec62, Noida, Uttar Pradesh India.

Division of Biochemistry, Galgotias University, Plot No.2, Sector 17-A Yamuna Expressway, Gautam Buddh Nagar, Uttar Pradesh India.

出版信息

J Food Sci Technol. 2017 Jun;54(7):2085-2093. doi: 10.1007/s13197-017-2647-z. Epub 2017 May 10.

Abstract

Polyphenol oxidase from corm was purified (5.54 fold) by acetone precipitation and ion exchange chromatography. Electrophoretic-blot analysis revealed that the molecular weight of enzyme is 40 kDa. Enzyme preferred catechol as substrate and showed optimum activity at pH 6 and 35 °C with Km of 17.56 mM and Vmax 0.023 U/ml/min. Thermal inactivation kinetics study showed that 5.68% inactivation was achieved at 75 °C within 40 min. The half-life time of PPO is between 126.00 and 32.54 min. The activation energy (Ea) and Z values are 64.22 kJ/mol and 34.01 °C, respectively. The inhibitory action of L-ascorbic acid, sodium azide, NaCl, CaCl, ZnSO and EDTA were studied. Thermodynamic studies indicated a non-spontaneous (ΔG > 0) and endothermic reaction process. Other thermodynamic activation parameters, such as Ea, ΔH, ΔG and ΔS, were also studied. polyphenol oxidase can find application in baking industry.

摘要

通过丙酮沉淀和离子交换色谱法对来自球茎的多酚氧化酶进行了纯化(5.54倍)。电泳印迹分析表明该酶的分子量为40 kDa。该酶优先选择儿茶酚作为底物,在pH 6和35℃时表现出最佳活性,Km为17.56 mM,Vmax为0.023 U/ml/min。热失活动力学研究表明,在75℃下40分钟内失活率达到5.68%。PPO的半衰期在126.00至32.54分钟之间。活化能(Ea)和Z值分别为64.22 kJ/mol和34.01℃。研究了L-抗坏血酸、叠氮化钠、NaCl、CaCl、ZnSO和EDTA的抑制作用。热力学研究表明这是一个非自发的(ΔG > 0)吸热反应过程。还研究了其他热力学活化参数,如Ea、ΔH、ΔG和ΔS。多酚氧化酶可应用于烘焙工业。

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