An efficient method to eliminate the protease activity contaminating commercial bovine pancreatic DNase I
| Autor: | Tien Le, Hyung Jong Jin, Hak Jin Lee |
|---|---|
| Rok vydání: | 2015 |
| Předmět: |
Proteases
Autolysis (biology) medicine.medical_treatment Biophysics Biology Protein degradation Biochemistry chemistry.chemical_compound Protein purification Escherichia coli medicine Animals Deoxyribonuclease I Molecular Biology Serine protease Chromatography Protease fungi Cell Biology chemistry biology.protein Cattle PMSF Peptide Hydrolases Phenylmethylsulfonyl Fluoride |
| Zdroj: | Analytical Biochemistry. 483:4-6 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2015.04.030 |
| Popis: | A method was developed to eliminate the proteases contaminating commercial DNase I, which can cause degradation of target protein during the purification process. Bio Basic DNase stock solution (in Tris–HCl buffer [pH 8.0] containing 5 mM CaCl2) was first incubated at 50 °C to generate autolysis of proteases and zymogens, leading to a significant reduction in protease activity while preserving DNase activity. The residual protease activity was completely inhibited by further incubation with 2 mM PMSF (phenylmethylsulfonyl fluoride) or 2× S8830 inhibitor cocktail. This approach could be readily applicable to eliminate the protease activity in any DNase products or during the preparation of commercial DNase. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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