School of Environment, Harbin Institute of Technology, Harbin 150090, China.
School of Marine Science and Technology, Harbin Institute of Technology, Weihai 264209, China.
Mar Drugs. 2020 Dec 4;18(12):618. doi: 10.3390/md18120618.
R-phycoerythrin (R-PE), a marine bioactive protein, is abundant in with high protein content. In this study, R-PE was purified using a deep eutectic solvents aqueous two-phase system (DES-ATPS), combined with ammonium sulphate precipitation, and characterized by certain techniques. Firstly, choline chloride-urea (ChCl-U) was selected as the suitable DES to form ATPS for R-PE extraction. Then, single-factor experiments were conducted: the purity (A/A) of R-PE was 3.825, and the yield was 69.99% () under optimal conditions (adding 0.040 mg R-PE to ChCl-U (0.35 g)/KHPO (0.8 g/mL, 0.5 mL) and extracting for 20 min). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results revealed that the purified R-PE contained three main bands. One band was presented after purification in native-PAGE. The UV-vis spectra showed characteristic absorption peaks at 495, 540, and 565 nm. R-PE displayed an emission wavelength at 570 nm when excited at 495 nm. All spectra results illustrated that the structure of R-PE remained unchanged throughout the process, proving the effectiveness of this method. Transmission electron microscope (TEM) showed that aggregation and surrounding phenomena were the driving forces for R-PE extraction. This study could provide a green and simple purification method of R-PE in drug development.
藻红蛋白（R-PE）是一种海洋生物活性蛋白，在 中含量丰富，且蛋白质含量高。本研究采用深共晶溶剂双水相体系（DES-ATPS）结合硫酸铵沉淀法对 R-PE 进行纯化，并采用一定的技术对其进行了表征。首先，选择氯化胆碱-尿素（ChCl-U）作为合适的 DES 来形成用于 R-PE 提取的 ATPS。然后进行单因素实验：在最佳条件下（向 ChCl-U（0.35 g）/KHPO（0.8 g/mL，0.5 mL）中加入 0.040 mg R-PE 并提取 20 min），R-PE 的纯度（A/A）为 3.825，收率为 69.99%（）。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）结果表明，纯化后的 R-PE 含有 3 条主要条带。在天然-PAGE 中纯化后出现一条带。紫外可见光谱在 495、540 和 565nm 处显示出特征吸收峰。当以 495nm 激发时，R-PE 显示出 570nm 的发射波长。所有光谱结果表明，R-PE 的结构在整个过程中保持不变，证明了该方法的有效性。透射电子显微镜（TEM）表明，聚集和周围现象是 R-PE 提取的驱动力。本研究可为药物开发中 R-PE 的绿色、简单纯化方法提供参考。
