title: An efficient method to eliminate the protease activity contaminating commercial bovine pancreatic DNase I.
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.
authors.affiliation: Department of Bioscience and Biotechnology, College of Natural Science, University of Suwon, Whasung City, Kyunggi-Do 445-743, Republic of Korea.
authors.forename: Tien
authors.initials: T
authors.lastname: Le
authors.affiliation: Department of Life Science, Korea University Graduate School, Seoul 136-713, Republic of Korea.
authors.forename: Hak Jin
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Department of Bioscience and Biotechnology, College of Natural Science, University of Suwon, Whasung City, Kyunggi-Do 445-743, Republic of Korea. Electronic address: hjjin@suwon.ac.kr.
authors.forename: Hyung Jong
authors.initials: HJ
authors.lastname: Jin
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002417:Cattle; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D010447:Peptide Hydrolases / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D003850:Deoxyribonuclease I; D010447:Peptide Hydrolases
keywords: Deoxyribonuclease (DNase); Protease inhibition; Protein degradation; Protein purification; Serine protease; Zymogen