Separation and Purification of Lipase using Reverse Micellar Extraction: Optimization of Conditions by Response Surface Methodology

Downstream processing of lipase involving
reverse micellar extraction of lipase using cationic surfactant
cetyltrimethylammonium bromide (CTAB) was investigated.
Effect of various process parameters on both forward and
backward extraction of lipase from crude extract was
studied to optimize its yield and purity. Complex interaction
of salt concentration (0.05~0.15M), surfactant concentration
(0.10~0.30 M), and pH (6.0~9.0) for forward extraction, as
well as, salt concentration (0.5~1.5 M) and pH (6.0~9.0)
for backward extraction have been studied using response
surface methodology. Optimum processing conditions,
namely, salt concentration 0.16M, surfactant concentration
0.20 M, and pH 9.0 for forward extraction, as well as, salt
concentration 0.80 M and pH 7.23 for backward extraction,
fulfill the conditions to obtain activity recovery of lipase
≥78% and purification factor of lipase ≥4.0. The study
demonstrated that response surface methodology can be
used for optimization of the conditions for reverse micellar
extraction of lipase.