Partial Purification and Characterization of Fibrinolytic Enzymes from Yellow Mealworm
| Autor: | Ming Xing Huang, Ya Li Han, Ya Xiong Chen, Yun Ye |
|---|---|
| Rok vydání: | 2012 |
| Předmět: |
chemistry.chemical_classification
Mealworm Chromatography biology Molecular mass Metal ions in aqueous solution Ion chromatography Size-exclusion chromatography Bioengineering biology.organism_classification Biochemistry Analytical Chemistry Enzyme chemistry Drug Discovery Molecular Medicine Ammonium sulfate precipitation Phenylmethylsulfonyl Fluoride |
| Zdroj: | International Journal of Peptide Research and Therapeutics. 18:153-161 |
| ISSN: | 1573-3904 1573-3149 |
| DOI: | 10.1007/s10989-012-9288-x |
| Popis: | Two proteins with fibrinolytic activity were partially purified from yellow mealworm (Tenebrio molitor) by ammonium sulfate precipitation between 30 and 70% saturation, gel filtration on Sephacryl-S200-HR, ion exchange chromatography on DEAE-Sepharose-FF and metal chelate on Cu–HiTrap–IMAC–FF, but the enzymes had not been completely separated from each other. The two partially purified fibrinolytic enzymes were designated as TMFE-I and TMFE-II (Tenebrio molitor fibrinolytic enzyme) with molecular weights of 27.5 and 24.9 kDa by SDS-PAGE individually. The partially purified solution of TMFE-I and TMFE-II was considerably stable in the range of pH 5–10 and characterized by pH optimum of the enzymatic activity at 8.0. Thermal stability of TMFE was excellent at 45°C and below. The K M value was 0.26 mM for amidolysis of Bz–Arg–pNA. According to inhibitor analysis by fibrin plate method, phenylmethylsulfonyl fluoride and tosyl-lysine chloromethyl ketone inactivated TMFE almost completely, but trans-(epoxysuccinyl)-l-leucylamino-4-guanidinobutane (E-64) and EDTA had little effect on their fibrinolytic activity. According to metal ion analysis by fibrin plate method, the effect of metal ions on activity of TMFE showed a great difference. Na+, K+ and Zn2+ had little effect on the activity of TMFE. Mg2+ and Cu2+ showed inhibition effect on the fibrinolytic activity of TMFE, but Ca2+ increased the fibrinolytic activity of TMFE at final concentration varying from 0 to 30 mM. |
| Databáze: | OpenAIRE |
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