Downstream Processing of Soy Hull Peroxidase Employing Reverse Micellar Extraction.

Phase transfer studies were conducted to
evaluate the solubilization of soy hull peroxidase (SHP) in
reverse micelles formed in isooctane/butanol/hexanol using
the cationic surfactant cetyltrimethylammonium bromide
(CTAB). The effect of various parameters such as pH,
ionic strength, surfactant concentration of the initial aqueous
phase for forward extraction and buffer pH, type and
concentration of salt, concentration of isopropyl alcohol
and volume ratio for back extraction was studied to
improve the efficiency of reverse micellar extraction. The
active SHP was recovered after a complete cycle of forward
and back extraction. A forward extraction efficiency
of 100%, back extraction efficiency of 36%, overall
activity recovery of 90% and purification fold of 4.72 were
obtained under optimised conditions. Anionic surfactant
sodium bis (2-ethylhexyl) sulfosuccinate (AOT) did not
yield good results under the conditions studied. The phase
transfer of soy hull peroxidase was found to be controlled
by electrostatic and hydrophobic interactions during
forward and back extraction respectively.