Xu Yuqin, Zeng Bin, Xiao Shuangling, Wang Di, Liu Yang, Chen Shengchang, Teng Jie
Department of Tea Science, College of Agriculture, Jiangxi Agricultural University, Nanchang 330045, China.
Suichuan Tea Research Institute, Ji'an 343009, China.
Food Chem X. 2024 Aug 7;23:101720. doi: 10.1016/j.fochx.2024.101720. eCollection 2024 Oct 30.
In this study, tea polyphenol oxidase (PPO) was purified via three-phase partitioning (TPP) using a deep eutectic solvent (DES) instead of -butanol. First, the properties of 13 types of synthesized DESs were characterized, and DES-7 (thymol/dodecanoic acid) was selected as the best alternative solvent. The process parameters were optimized using response surface methodology. The experimental results revealed that when the (NH)SO concentration, DES to crude extract ratio, extraction time, and pH were 41%, 0.5:1, 75 min, and 5.6, respectively, the recovery and purification fold of tea PPO were 78.44% and 8.26, respectively. SDS-PAGE and native-PAGE were used to analyze the PPO before and after purification of the TTP system, and the molecular weight and purification effect of PPO were detected. Moreover, the DES could be recovered and recycled. The results indicate an environmentally friendly and stable DES, and provide a reference for the large-scale application of TPP to extract PPO.
在本研究中，使用深共熔溶剂（DES）而非叔丁醇通过三相分离（TPP）法纯化了茶多酚氧化酶（PPO）。首先，对13种合成DES的性质进行了表征，并选择DES-7（百里香酚/十二烷酸）作为最佳替代溶剂。采用响应面法对工艺参数进行了优化。实验结果表明，当硫酸铵浓度、DES与粗提物的比例、提取时间和pH分别为41%、0.5:1、75分钟和5.6时，茶PPO的回收率和纯化倍数分别为78.44%和8.26。使用SDS-PAGE和非变性PAGE分析了TTP系统纯化前后的PPO，并检测了PPO的分子量和纯化效果。此外，DES可以回收再利用。结果表明DES对环境友好且稳定，为TPP大规模应用提取PPO提供了参考。
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