Pseudomonas sp. S4-14가 생산하는 Alkaline Lipase의 정제 및 효소학적 성질

The strain S4-14 which produced alkaline lipase and had resistance against linear alkylbenzene sulfonate was isolated from soil or water samples. The isolated strain S4-14 was identified a species belong to Pseudomonas. Alkalin lipase secreted by Pseudomonas sp. S4-14 was purified by ammonium sulfate precipitation procedure follwed by DEAE-Cellulose, DEAE-Sepharose and gel filtration chromatohraphies with 995.15 U/mg protein and 16.1% yield. The molecular weight of the enzyme was estimated to be 65,000 dalton by SDS-PAGE. The optimum pH and temperature of the purified enzyme was 10.5 and 45$\circ $C, respectively. The emzyme was stable at 45$\circ $C for 1 hr and in a pH range from 8.0 to 12.0 for 24 hr at 4$\circ $C. The activity of lipase was enhanced by Ca$^{2+}$ while inhibited strongly by Pb$^{2+}$, Zn$^{2+}$ or Fe$^{3+}$. The activity of lipase was inactivated about 50~60% in the presence of 50 mg/l linear alkylbenzene sulfonate, $\alpha $-olefin sulfonate, alcohol ethoxylate or perborate.