Optimization of Lipase Immobilization

Pseudomonas aeruginosa BBRC-10036 was used for lipase production. The organism secreted the enzyme extracellulary. In order to purify the enzyme, precipitation was done first, and then this lipase has been purified by Ion exchange Chromatography leading to 2.3-fold purification and 11.47% recovery. Lipase from P.aeruginosa was entrapped into Ca-alginate gel beads and effect of independent variables such as alginate concentration (%w/v), CaCl2 concentration (M) and enzyme load (%v/v) on immobilization yield and activity of immobilized enzyme were investigated. Media optimization for immobilization of lipase was carried out by Response Surface Methodology. The optimum conditions were as follows: sodium alginate concentration 2.5% (w/v), calcium chloride concentration 2.5(M) and enzyme load 50% (v/v). At those conditions, the highest immobilization yield and the optimum activity of immobilized enzyme, respectively, obtained were 93.65% and 2.64 unit/gr(IME).