Authors:
Chunming Tan, Gang Yu, Yong Xue, Lin Wang and Shaoling Yang, China |
Abstract:
Using Tubificidae as the material was purified by historrhexis, salting out, ultra filtration, Sephadex G-50 gel filtration chromatography and DEAE Sepharose Fast Flow anion-exchange chromatography, acquired pure product of lumbrokinase. The activity of the lumbrokinase was detected by agarose-fibrin plate method. After purification by anion-exchange chromatography, the purity of the lumbrokinase with a specific activity of 5630.4 U/mg and increased 25.2-fold over the purity of the supernatant. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the molecular weight of the purified lumbrokinase distributed at the range of 15-29 kDa, namely, 15 kDa, 27 kDa, 29 kDa. The optimal temperature and pH for the killing activity of the purified killer toxin were 36 0C and 8.0, respectively, and the killing activity was stable over a temperature range of 0-50 0C and pH range of 7.0-10.0. Moreover, the ions increased the enzyme activity of K+, Na+, Mg2+, Ca2+ and Fe3+, while Pb2+ and Hg2+ have inhibitory effect. In addition, we used microorganism depressed method ( cup method) to test the anti-microbial? effects, we found that lumbrokinase can nhibit the growth of Staphylococcus aureus, Escherichia coli, klebsiella pneumoniae, Candida albicans and Bacillus subtilis. Especially for klebsiella pneumoniae and Candida albicans, the effects were extremely significant. |
