Rapid extraction and purification of lumbrokinase from Lumbricus rubellus using a hollow fiber membrane and size exclusion chromatography.
OBJECTIVE
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.
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China. futm@njucm.edu.cn.|Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, Nanjing, 210029, China. futm@njucm.edu.cn.
Tingming
T
Fu
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Fengyun
F
Yang
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Huaming
H
Zhu
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Huaxu
H
Zhu
School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China.
Liwei
L
Guo
D000818:Animals; D002850:Chromatography, Gel / Q000379:methods*; D010450:Endopeptidases / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods*; D008566:Membranes; D009835:Oligochaeta / Q000201:enzymology*; D013997:Time Factors
D010450:Endopeptidases; C083421:lumbrokinase
Earthworms; Fibrinolytic activity; Hollow fiber membrane; Lumbricus rubellus; Lumbrokinase