萝卜
化学
硫酸铵
色谱法
萃取(化学)
深共晶溶剂
十二烷基硫酸钠
过氧化物酶
蔗糖
盐(化学)
响应面法
核化学
共晶体系
有机化学
植物
酶
合金
生物
作者
Yuxiao Hu,Xueying Chen,Zhijian Tan
标识
DOI:10.1016/j.seppur.2023.124353
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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