芽孢杆菌WF63脂肪酶分离纯化及酶学性质  

Purification and Characterization of Lipase from Bacillus sp. WF63

作　　者：黄鹭强[1] 谢必峰[1] 黄建忠[1] 吴松刚[1] 

机构地区：[1]福建师范大学生命科学学院,福建福州350108

出　　处：《云南民族大学学报（自然科学版）》2010年第4期290-293,共4页Journal of Yunnan Minzu University:Natural Sciences Edition

基　　金：国家自然科学基金(30870545);福建省教育厅科技项目(JA05199)

摘　　要：芽孢杆菌WF63脂肪酶发酵上清液经硫酸铵沉淀、Sephaeryl$200柱层析、CMSephrose FF柱层析，得到纯化的脂肪酶，纯化倍数为9．45倍，活性回收率为4．27％．对该酶性质研究表明：该酶水解橄榄油的最适温度为50℃，最适pH为8．0，在60℃以下和pH4～10之间有很好的稳定性．K^＋，Mgn对该脂肪酶的水解活性有促进作用，而Na^＋，Mn^2＋，Ca^2＋对脂肪酶则没有显著的促进作用，Fe^2＋对酶活的抑制作用不明显．浓度为0．1％时，Triton X-100对脂肪酶活力有明显促进作用；Brij 35，Tween-80对酶活力的影响不明显；离子型表面活性剂SDS则表现出抑制作用．A lipase from Bacillus sp. WF63 was purified to homogeneity by using annonium sulfate precipitation, Sephaeryl S200 gel filtration and CM Sephrose Fast Flow ion exchange chromatography. This purification protocol resulted in a 9.45 fold purification of lipase with 4.27% final yield, and the relative molecular weight of the enzyme was determined to be 32kw by SDS PAGE approximately. The optimum pH of the lipase was 8.0 and the optimum temperature was 50 ℃. The lipase was highly stable in the pH range from 4 to 10. K^＋ and Mg^2＋ stimulated lipolytic activity, Na^2＋ , Mn^2＋ , Ca^2＋ stimulated lipolytic activity, whereas, Fe2 ~ caused inhibition. Triton X - 100 stimulated lipolytic activity, Brij 35, Tween - 80 stimulated lipolytic activity, whereas, SDS caused inhibition under 1% surfactant.

关 键 词：芽孢杆菌 脂肪酶 分离纯化 性质 

分 类 号：Q55[生物学—生物化学]