交联重组枯草杆菌纤溶酶聚集体的制备及其性质研究  

Preparation and characterization of cross-linked recombinant BSFE aggregates

作　　者：潘延芳[1] 孔珊珊[1] 陈怡倩[1] 吴自荣[1] 王隆华[1] 

机构地区：[1]华东师范大学生命科学学院,上海200062

出　　处：《中国生化药物杂志》2007年第5期318-321,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：华东师范大学2005年"大夏"科研基金(KY2005-011)

摘　　要：目的制备交联重组枯草杆菌纤溶酶聚集体(CLEAs),对其酶学性质进行研究,提高重组枯草杆菌纤溶酶(rBSFE)的稳定性。方法通过微生物发酵,分离得天然rBSFE,经硫酸铵沉淀后,以戊二醛作为交联剂对其进行化学交联,得CLEAs。用常规方法测定各种因素对rBSFE稳定性的影响。结果在对耐热性、有机相中稳定性、对抗血清蛋白水解酶的能力的研究中,CLEAs均显示比游离酶更高的稳定性。在有机相N,N-二甲基甲酰胺中作用24 h后,CLEAs的活性下降速率为游离酶的50%。在小鼠体外血清中作用2.5 h后,游离酶活性下降56.3%,而CLEAs活性仅下降4.5%,稳定性显著高于游离酶。结论运用CLEA技术可以较大提高rBSFE的稳定性。Purpose To enhance the stability of recombinant fibrinolytic enzyme from Bacillus sp. N18 （rBSFE）. Methods rBSFE was obtained by microbe＇s zymosis, then immobilized in a simple and effective way by physical aggregation using a precipitant-ammonium sulfate, followed by chemical cross-linking via a bifunctional reagent-glutaraldehyde to form insoluble cross-linked recombinant BSFE aggregates （CLEAs）. The optimum way of preparing CLEAs was identified, and the enzyme property was Studied. Results Compared with those of free rBSFE, the thermal stability, stability in organic solvents, and resistance of CLEAs to the exogenous proteolysis were enhanced. After being stirred in N, N-Dimethylformamide for 24 hours, the declining activity rate of free rBSFE was twice of that of CLEAs. After being stirred in in vitro serum for 2.5 hours, the activity of free recombinant BSFE decreased by 56.3 %, while the one of CLEAs only decreased by 4.5 %. Conclusion The stability of rBSFE can be highly strengthened by CLEA techniques.

关 键 词：重组枯草杆菌纤溶酶 交联酶聚集体 固定化 

分 类 号：Q814.1[生物学—生物工程] Q55