Characterization of deoxyribonuclease I immobilized on magnetic hydrophilic polymer particles
- 5 July 2002
- journal article
- Published by Elsevier BV in Journal of Chromatography B
- Vol. 774 (1), 25-31
- https://doi.org/10.1016/s1570-0232(02)00163-0
Abstract
Magnetic bead cellulose particles and magnetic poly(HEMA–co–EDMA) microspheres with immobilized DNase I were used for degradation of chromosomal and plasmid DNAs. Magnetic bead particles were prepared from viscose and magnetite powder. Magnetic poly(HEMA–co–EDMA) microspheres were prepared by dispersion copolymerization of 2-hydroxyethyl methacrylate and ethylene dimethacrylate in the presence of magnetite. Divalent cations (Mg2+, Ca2+, Mn2+ and Co2+) were used for the activation of DNase I. A comparison of free and immobilized enzyme (magnetic bead particles) activities was carried out in dependence on pH and activating cation. The maximum of the activity of immobilized DNase I was shifted to lower pH compared with free DNase I. DNase I immobilized on magnetic bead cellulose was used 20 times in the degradation of chromosomal DNA. Its residual activity was influenced by the nature of activating divalent cation. The immobilized enzyme with decreased activity was reactivated by Co2+ ions.Keywords
This publication has 14 references indexed in Scilit:
- Influence of Surface Hydrophobic Groups on the Adsorption of Proteins onto Nonporous Polymeric Particles with Immobilized Metal IonsJournal of Colloid and Interface Science, 2001
- Properties of RNase A Immobilized on Magnetic Poly(2‐hydroxyethyl methacrylate) MicrospheresBiotechnology Progress, 2001
- Carp liver actin: isolation, polymerization and interaction with deoxyribonuclease I (DNase I)Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, 1999
- Oriented immobilization of restriction endonuclease ecoriJournal of Molecular Recognition, 1996
- Capillary enzymophoresis of nucleic acid fragments using coupled capillary electrophoresis and capillary enzyme microreactors having surface-immobilized RNA-modifying enzymesElectrophoresis, 1995
- DNase activity of micrococcal endonuclease covalently immobilized on nylon and polystyreneEnzyme and Microbial Technology, 1993
- Immobilized NucleasesCritical Reviews in Biotechnology, 1993
- The Isolation of Actin from Pea Roots by DNase I Affinity ChromatographyPlant Physiology, 1992
- A coupled transcription—translation system derived from Escherichia coli: The use of immobilized deoxyribonuclease to eliminate endogenous DNAFEBS Letters, 1976
- CRYSTALLINE DESOXYRIBONUCLEASEThe Journal of general physiology, 1950