Cytochrome c is a small water-soluble protein that is abundantly found in the mitochondrial intermembrane space of microorganism, plants and mammalians. Ionic liquids (ILs)-based aqueous two-phase electrophoresis system (ATPES) was introduced in this study to investigate the partition efficiency of cytochrome c to facilitate subsequent development of two-phase electrophoresis for the separation of cytochrome c from microbial fermentation. The 1-Hexyl-3-methylimidazolium bromide, (C6mim)Br and potassium citrate salt were selected as the phase-forming components. Effects of phase composition; position of electrodes; pH and addition of neutral salt on the partition efficiency of cytochrome c in the ATPES were evaluated. Highest partition coefficient (K = 179.12 ± 0.82) and yield of cytochrome c in top phase (YT = 99.63% ± 0.00) were recorded with IL/salt ATPES composed of 30% (w/w) (C6mim)Br and 20% (w/w) potassium citrate salt of pH 7 and 3.0% (w/w) NaCl addition with anode at the bottom phase and cathode at the top phase. The SDS-PAGE profile revealed that cytochrome c with a molecular weight of 12 kDa was preferably partitioned to the IL-rich top phase. Present findings suggested that the single-step ATPES is a potential separation approach for the recovery of cytochrome c from microbial fermentation. Graphical Abstract.

中文翻译：

---

基于离子液体的水两相电泳对细胞色素c分配的效率

细胞色素c是一种小的水溶性蛋白质，广泛存在于微生物，植物和哺乳动物的线粒体膜间空间中。本研究引入基于离子液体（ILs）的水性两相电泳系统（ATPES），以研究细胞色素c的分配效率，以促进随后用于从微生物发酵分离细胞色素c的两相电泳的发展。选择溴化1-己基-3-甲基咪唑鎓，（C6mim）Br和柠檬酸钾盐作为相形成组分。相组成的影响；电极位置；评估了pH和中性盐的添加对ATPES中细胞色素c分配效率的影响。最高分配系数（K = 179.12±0.82）和最高阶段的细胞色素c产量（YT = 99.63％±0）。用由30％（w / w）（C6mim）Br和20％（w / w）pH 7的柠檬酸钾盐和3.0％（w / w）NaCl组成的IL /盐ATPES记录00）。底部相和顶部相的阴极。SDS-PAGE图谱表明，分子量为12kDa的细胞色素c优选分配至富含IL的顶层。目前的发现表明，单步ATPES是从微生物发酵中回收细胞色素c的潜在分离方法。图形概要。目前的发现表明，单步ATPES是从微生物发酵中回收细胞色素c的潜在分离方法。图形概要。目前的发现表明，单步ATPES是从微生物发酵中回收细胞色素c的潜在分离方法。图形概要。