数字农科院2.0

Three-phase partitioning constructed by pH-responsive deep eutectic solvents and sugars for purification of radish (Raphanus sativus L.) peroxidase

文献类型： 外文期刊

作者： Yuxiao Hu;Xueying Chen;Zhijian Tan

作者机构：

关键词： pH-responsive deep eutectic solvent;Radish peroxidase;Recyclable extractant;Sugaring-out effect;Three-phase partitioning

期刊名称： Separation and Purification Technology

ISSN： 1383-5866

年卷期： 2023 年 322 卷

页码：

收录情况： SCIE（2023版） ; ; EI（2023版）

摘要： Ammonium sulfate and t-butanol are widely used in conventional three-phase partitioning (TPP); however, their use is not eco-friendly. In this study, a novel TPP was developed using pH-responsive deep eutectic solvents (DESs) and sugars as substitutes for t-butanol and ammonium sulfate, respectively. This TPP was further used for the separation and purification of radish (Raphanus sativus L.) peroxidase (POD). DES-3 (dodecanoic acid: decanoic acid = 1:1) and sucrose were selected as optimal phase-forming components. Single-factor experiments were performed to investigate the factors influencing the TPP process, and three major factors (sucrose concentration, extraction time, and extraction temperature) were optimized by response surface methodology (RSM). Under the optimized conditions, the POD recovery and purification fold reached 101.66% and 11.56, respectively. Cyclic experiments confirmed the retrievability of DESs. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) confirmed that the novel TPP had better purification efficiency than the traditional TPP. The method proposed in this study is more efficient than conventional TPP, and it does not require the use of organic solvents and high concentration salt solutions. Therefore, this method is promising in the application of TPP.

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