Tingming Fu1,2, Fengyun Yang3, Huaming Zhu3, Huaxu Zhu3, Liwei Guo3. 1. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China. futm@njucm.edu.cn. 2. Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing, 210029, China. futm@njucm.edu.cn. 3. School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Abstract
OBJECTIVES: To develop a purification process using hollow fiber membrane separation combined with size exclusion chromatography for the extraction of lumbrokinase from earthworms (Lumbricus rubellus). RESULTS: To extract the protein, the earthworms were first homogenized for 10 min, to produce ultrafine particles. Polyether sulfone hollow fiber membranes with MW cut offs of 50 and 6 kDa were used for initial purification of the crude extract. Further purification was carried out on a Sephadex G-75 column, and yielded three fractions of high purity protein. One of these fractions showed fibrinolytic activity. CONCLUSIONS: Three samples of high purity protein were obtained and one protein (LK1) showed strong fibrinolytic activity. The method has higher purification efficiency in comparison with existing methods.
OBJECTIVES: To develop a purification process using hollow fiber membrane separation combined with size exclusion chromatography for the extraction of lumbrokinase from earthworms (Lumbricus rubellus). RESULTS: To extract the protein, the earthworms were first homogenized for 10 min, to produce ultrafine particles. Polyether sulfone hollow fiber membranes with MW cut offs of 50 and 6 kDa were used for initial purification of the crude extract. Further purification was carried out on a Sephadex G-75 column, and yielded three fractions of high purity protein. One of these fractions showed fibrinolytic activity. CONCLUSIONS: Three samples of high purity protein were obtained and one protein (LK1) showed strong fibrinolytic activity. The method has higher purification efficiency in comparison with existing methods.