Reverse micellar extraction for downstream processing of lipase: Effect of various parameters on extraction

Reversemicellar extraction of lipase using cationic surfactant cetyltrimethylammonium bromide (CTAB)
was investigated. The effect of various process parameters on both forward and backward extraction of
lipase from crude extract was studied to optimize its yield and purity. Forward extraction of lipase was
found to be maximum using Tris buffer at pH 9.0 containing 0.10 M NaCl in aqueous phase and 0.20 M
CTAB in organic phase consisting of isooctane, butanol and hexanol. In case of backward extraction,
lipase was extracted from the organic phase to a fresh aqueous phase in 0.05 M potassium phosphate
buffer (pH 7.0) containing 1.0 MKCl. The activity recovery, extraction efficiency and purification factor of
lipase were found to be 82.72%, 40.27% and 4.09-fold, respectively. The studies also indicated that the
organic phase recovered after back extraction could be reused for the extraction of lipase from crude
extract.