Lactoperoxidase partitioning from whey using the reverse micelles of non‐ionic/ionic mixed surfactants: Improvement of back extraction

Abstract

AbstractThe reverse micellar system formed with a mixture of ionic and non‐ionic surfactants, AOT (Sodium bis[2‐ethyl hexyl] sulfosuccinate)/Tween 80 in isooctane, was studied for the selective extraction of Bovine Lactoperoxidase (LP) from acid whey. The effect of pH and ionic strength of acid whey and surfactant concentrations were studied and a maximum of 86% LP was extracted from acid whey at pH 9.5 with the addition of 0.2 M KCl to the reverse micelles formed with AOT (115 mM)/Tween 80 (23 mM). The back extraction of LP was studied at different aqueous stripping phase pH, ionic strength and concentration of counter‐ionic surfactant Cetyltrimethylammonium bromide (CTAB). The back extraction of 112% with 80% LP recovery was achieved when the stripping phase pH was 10.5 with 1.5 M KCl and 60 mM CTAB. The antimicrobial activity of the extracted LP showed reduction in colony‐forming units of S. aureus.Novelty impact statementThe reverse micelles formed with AOT/Tween 80 surfactant mixture minimize the pH‐dependent denaturation of LP and widen the pH window (7.5–9.5) for LP extraction. The back extraction of LP from the reverse micellar phase to aqueous stripping phase was improved by the addition of CTAB as counter‐ionic surfactant. A purification fold of 11.26 achieved with minimal loss in activity of LP by retaining the native Antimicrobial characteristic.