Abstract
Serratia marcescens are well-known Gram-negative bacteria capable of excreting extracellular hydrolases, such as the nuclease—an enzyme that catalyzes hydrolysis of both DNA and RNA chains with a high efficiency. The nuclease has a number of attractive properties for use in biotechnology industry—outstanding enzymatic activity and low production cost. The existing protocols for purification of nuclease yield only limited amounts of protein and require complicated multistage procedures. Here, we report a chromatographic protocol for elegant one-step purification procedure resulting in a pure homogenous enzyme, as confirmed by gel electrophoresis and mass spectrometry.
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References
Benedik, M. J., & Strych, U. (1998). Serratia marcescensand its extracellular nuclease. FEMS Microbiology Letters, 165(1), 1–13. doi:10.1111/j.1574-6968.1998.tb13120.x.
Sheng-Fowler, L., Lewis, A. M., Peden, K. (2009). Quantitative determination of the infectivity of the proviral DNA of a retrovirus in vitro: evaluation of methods for DNA inactivation. Biologicals, 37(4), 259–269. doi:10.1016/j.biologicals.2009.04.002.
Hagen, A. J., Aboud, R. A., Dephillips, P. A., Oliver, C. N., Orella, C. J., Sitrin, R. D. (1996). Use of nuclease enzyme in the purification of VAQTA, a hepatitis A vaccine. Biotechnology and Applied Biochemistry, 23(Pt 3), 209–215.
Li, S.-M., Bai, F.-L., Xu, W.-J., Yang, Y.-B., An, Y., Li, T.-H., et al. (2014). Removing residual DNA from Vero-cell culture-derived human rabies vaccine by using nuclease. Biologicals, 42(5), 271–276. doi:10.1016/j.biologicals.2014.06.005.
Huyghe, B. G., Liu, X., Sutjipto, S., Sugarman, B. J., Horn, M. T., Shepard, H. M., et al. (1995). Purification of a type 5 recombinant adenovirus encoding human p53 by column chromatography. Human Gene Therapy, 6(11), 1403–1416. doi:10.1089/hum.1995.6.11-1403.
Drittanti, L., Jenny, C., Poulard, K., Samba, A., Manceau, P., Soria, N., et al. (2001). Optimised helper virus-free production of high-quality adeno-associated virus vectors. The Journal of Gene Medicine, 3(1), 59–71. doi:10.1002/1521-2254(2000)9999:9999<::AID-JGM152>3.0.CO;2-U.
Slepushkin, V., Chang, N., Cohen, R., Gan, Y., Jiang, B., Deausen, E., et al. (2003). Large-scale purification of a lentiviral vector by size exclusion chromatography or mustang Q ion exchange capsule. BioProcessing Journal, 2(5), 89–95.
Pedersen, J., Filimonova, M., Roepstorff, P., Biedermann, K. (1993). Characterization of Serratia marcescens nuclease isoforms by plasma desorption mass spectrometry. Biochimica et Biophysica Acta, 1202(1), 13–21.
Acknowledgments
This work was supported by Program for Competitive Growth of Kazan Federal University. Emil Bulatov was supported by grant 16-34-60213 mol_a_dk from the Russian Foundation for Basic Research. Authors thank Dr. Julia Romanova for help with mass spectrometry experiments.
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Vafina, G., Bulatov, E., Zaynutdinova, E. et al. A One-Step Protocol for Chromatographic Purification of Non-recombinant Exogenous Bacterial Enzyme: Nuclease of Serratia marcescens . BioNanoSci. 6, 335–337 (2016). https://doi.org/10.1007/s12668-016-0226-9
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DOI: https://doi.org/10.1007/s12668-016-0226-9