微生物产弹性蛋白酶制取方法的探讨

Approach on Preparative Method of Elastase Produced by Microorganism

目的探讨微生物产弹性蛋白酶的制取方法。方法将产弹性蛋白酶的枯草芽孢杆菌接种到发酵培养基中,摇床发酵培养后,离心收集发酵液,并经过硫酸铵盐析、DEAE-Sephadex A-25阴离子交换层析、SephadexG-75凝胶过滤层析等步骤纯化弹性蛋白酶。结果经过上述纯化步骤,弹性蛋白酶纯化倍数达到了33倍,回收率达到12%,酶蛋白比活达到了528 U/mg。结论此纯化工艺较为理想,获得了较纯的酶蛋白。

Objective To investigate the preparative method of elastase produced by microorganism. Methods A elastase-producing strain of Bacillus subtilis was inoculated into fermentation medium and cultivated on a rotary shaker, The broth was collected by centrifugation. The purification of elastase was achieved by combination methods of （NH4）2SO4 fractionation, anion exchange chromatography on DEAE-Sephadex A-25 and gel filtration chromatography on Sephadex G-75. Results Through the purification procedures shown above, the purification multiple of elastase was 33, the recovery rate was up to 12 % and the specific activity was about 528 U/mg. Conclusion The purer elastase can be obtained by this ideal purification technology.