Abstract |
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 5mM 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 2mM 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.
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Authors | Tien Le, Hak Jin Lee, Hyung Jong Jin |
Journal | Analytical biochemistry
(Anal Biochem)
Vol. 483
Pg. 4-6
(Aug 15 2015)
ISSN: 1096-0309 [Electronic] United States |
PMID | 25944416
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Copyright | Copyright © 2015 Elsevier Inc. All rights reserved. |
Chemical References |
- Deoxyribonuclease I
- Peptide Hydrolases
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Topics |
- Animals
- Biochemistry
(methods)
- Cattle
- Deoxyribonuclease I
(metabolism)
- Escherichia coli
(metabolism)
- Peptide Hydrolases
(isolation & purification, metabolism)
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