Background

The purification of keratinase is essential to enhance the catalytic efficiency of keratinase enzyme for industrial applications. Biphasic system incorporated with electric fields is an effective purification tool for the recovery of biomolecules. Ionic liquids (IL) with good conductivity are ideal phase-forming components for biphasic system which promotes the separation of charged biomolecules via electromigration.

Methods

This study aimed to evaluate the effect of electric fields on enhanced recovery of Kytococcus sedentarius TWHKC01 keratinase using IL-based biphasic system.

Findings

Keratinase was recovered with biphasic system incorporated with anode in salt-rich bottom phase and cathode in IL-rich top phase operated at 5 V for 5 min. A total of 95.82% ± 0.17 of keratinase was attained with the biphasic system at pH 9 comprising of 20% (w/w) 1‑butyl‑3-methylimidazolium tetrafluoroborate ((Bmim)BF₄) and 25% (w/w) ammonium sulphate loaded with 15% (w/w) K. sedentarius TWHKC01 crude feedstock. The selectivity (S) and purification fold (PF_T) of keratinase were increased from 7.25 ± 1.50 to 10.87 ± 0.61 and 5.94 ± 0.22 to 8.22 ± 0.44, respectively with the incorporation of electrodes, demonstrating the efficiency of electric fields in enhancing the purity of recovered keratinase.

中文翻译：

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将电场结合到基于离子液体的水性双相系统中以增强细胞外久坐不动杆菌 TWHKC01 角蛋白酶的恢复

背景

角蛋白酶的纯化对于提高角蛋白酶在工业应用中的催化效率至关重要。与电场结合的双相系统是回收生物分子的有效净化工具。具有良好导电性的离子液体 (IL) 是双相系统的理想相形成组分，可通过电迁移促进带电生物分子的分离。

方法

本研究旨在评估电场对使用基于 IL 的双相系统增强久坐Kytococcus sentarius TWHKC01 角蛋白酶恢复的影响。

发现

角蛋白酶用双相系统回收，其中阳极位于富含盐的底部相中，阴极位于富含 IL 的顶部相中，在 5 V 下运行 5 分钟。使用由 20% (w/w) 1-丁基-3-甲基咪唑鎓四氟硼酸盐 ((Bmim)BF ₄ ) 和 25% (w/w)组成的 pH 9 的双相系统获得总共 95.82% ± 0.17 的角蛋白酶负载有 15% (w/w) K. sedentarius TWHKC01 原油原料的硫酸铵。加入电极后，角蛋白酶的选择性（S）和纯化倍数（P _T）分别从 7.25 ± 1.50 增加到 10.87 ± 0.61 和 5.94 ± 0.22 到 8.22 ± 0.44，证明了电场在提高纯度方面的效率回收的角蛋白酶。