Incorporation of electric fields to ionic liquids-based aqueous biphasic system for enhanced recovery of extracellular Kytococcus sedentarius TWHKC01 keratinase

Abstract

BackgroundThe 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. MethodsThis study aimed to evaluate the effect of electric fields on enhanced recovery of Kytococcus sedentarius TWHKC01 keratinase using IL-based biphasic system. FindingsKeratinase 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 (PFT) 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.