Effect of Factors on Pepsin and Trypsin Fibrinolytic Activity and Deactivation of Fibrinolytic Activity
(2.Key Laboratory of Separation Engineering for Chinese Medicine Compound, Nanjing University of Chinese Medicine, Nanjing, China 210029)
(3.The First Clinical Medical College, Nanjing University of Chinese Medicine, Nanjing, China 210046)
(4.Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor, Nanjing University of Chinese Medicine, Nanjing, China 210023)
【Abstract】 Objective To investigate the method for study on the effect of factors on pepsin and trypsin fibrinolytic activity and deactivation of fibrinolytic activity and to eliminate the interference of pepsin and trypsin on the detection of crude protein fibrinolytic activity of Armadillidium vulgare (porcellio plasmin) in order to obtain the proteins or peptides which have the smaller molecular weight but higher titer during the pepsin and trypsin degradation. Methods To study the effect of pepsin and trypsin deactivation on pH value, temperature, metal ions, enzyme inhibitor, surfactant, and responsing fibrinolytic, the fiber fibrin plate assay was usede. The better enzyme deactivation process was obtained and used for studying the effect on the fibrinolytic activity of urokinase, lumbrokinase, and porcellio plasmin. Results All the pH value, temperature, metal ions, enzyme inhibitor, and surfactant have had an impact on pepsin and trypsin fibrinolytic activity. Among them the optimum deactivation of pepsin was pH 6.0–8.0, while the optimum deactivation of trypsin was mixed preparation with TLCK at the concentration of 25 mg/mL and EDTA at the concentration of 1 mmol/L. Conclusion This study has obtained the better enzyme deactivation process which could be used for the detection of fibrinolytic activity of pepsin and trypsin degradation product by fiber fibrin plate assay; the operation is simple, and the repeatability and stability are good.
【Keywords】 pepsin; trypsin; fibrinolytic activity; fiber fibrin plate assay; enzyme deactivation; urokinase; lumbrokinase; porcellio plasmin;
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