Abstract Keratinase is widely applied in the bioconversion of feather waste to produce value-added bioproducts. The wide applications of industrial keratinase have urged the advancement in producing keratinase with high productivity and yields. The efficiency of aqueous two-phase electrophoresis system (ATPES) to recover extracellular Kytococcus sedentarius TWHKC01 keratinase was investigated. The operation time of 10 min and voltage of 10 V were applied for the recovery of K. sedentarius TWHKC01 keratinase with the PEG 1000/sulfate ATPES. Keratinase was recovered in the PEG-rich phase of the ATPES consists of 23% (w/w) PEG 1000, 18.5% (w/w) ammonium sulfate of pH 9, 20% (w/w) crude feedstock and with 3% (w/w) of NaCl added. Highest partition coefficient (Ke) of 4.63 ± 0.17, recovery yield (YT) of 78.75% ± 0.61, and purification fold (PFT) of 3.89 ± 0.84 were obtained in the optimal ATPES coupled with cathode electrode in top phase and anode electrode in the bottom phase. The introduction of electric field to the ATPS has enhanced the selectivity (S) of the system towards the extracellular keratinase by two-fold from 2.79 ± 0.46 to 5.68 ± 0.43, demonstrating the efficiency of ATPES in separating the keratinase from crude feedstock when compared to conventional two-phase system.

中文翻译：

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聚合物/盐水溶液两相电泳系统回收细胞外久坐不动杆菌TWHKC01角蛋白酶的效率

摘要 角蛋白酶广泛应用于羽毛废料的生物转化，以生产高附加值的生物制品。工业角蛋白酶的广泛应用推动了生产具有高生产率和产量的角蛋白酶的进步。研究了水性两相电泳系统 (ATPES) 回收细胞外久坐不动杆菌 TWHKC01 角蛋白酶的效率。10 分钟的操作时间和 10 V 的电压用于使用 PEG 1000/硫酸盐 ATPES 回收 K. sedentarius TWHKC01 角蛋白酶。在 ATPES 富含 PEG 的相中回收角蛋白酶，该相由 23% (w/w) PEG 1000、18.5% (w/w) pH 9 的硫酸铵、20% (w/w) 粗原料和 3% (w/w) 的 NaCl 添加。最高分配系数 (Ke) 为 4.63 ± 0.17，回收率 (YT) 为 78.75% ± 0.61，纯化倍数 (PFT) 为 3。在与顶部相的阴极电极和底部相的阳极电极耦合的最佳 ATPES 中获得了 89±0.84。将电场引入 ATPS 后，系统对细胞外角蛋白酶的选择性 (S) 提高了两倍，从 2.79 ± 0.46 到 5.68 ± 0.43，证明了 ATPES 将角蛋白酶从粗原料中分离的效率与传统的两相系统。