Purification effect of artificial chaperone in the refolding of recombinant ribonuclease A from inclusion bodies |
| |
| Institution: | 1. Graduate Program in Biosciences, Life Science Division, University of Guanajuato Campus Irapuato-Salamanca. Irapuato, Guanajuato 36500, Mexico;2. Food Department, Life Science División, University of Guanajuato Campus Irapuato-Salamanca, Irapuato, Guanajuato 36500, Mexico;3. Department of Chemistry, Division of Natural and Exact Sciences, University of Guanajuato Campus Guanajuato, 36250, Mexico;4. Department of Biological Sciences, California Baptist University, 8432 Magnolia Avenue, Riverside, CA 92504, USA;5. Department of Entomology, University of California, Riverside, CA 92521, USA;1. Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2P5, Canada;2. School of Public Health, University of Alberta, Edmonton, Alberta T6G 2P5, Canada;3. Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Alberta T6G 2P5, Canada |
| |
| Abstract: | Artificial chaperone (AC) containing cetyltrimethylammonium bromide (CTAB) and β-cyclodextrin (β-CD) has been used to refold recombinant ribonuclease A (RNase A) from inclusion bodies (IBs). At low urea concentration (0.8 M), the AC could enhance the refolding yield of RNase A by effectively suppressing its intermolecular interaction-induced aggregation. As a result, 0.9 mg/mL RNase A could be 77% refolded, which was a 57% increase as compared to that without the AC. At high protein concentration range (0.9–2.3 mg/mL in total protein concentrations) and 1.6 M urea, CTAB selectively precipitated contaminant proteins distinctly, so a purification effect was achieved. For example, 1.5 mg/mL RNase A could be 62% refolded and recovered at a purity of 87%, which was a 34% increase in purity as compared to that in IBs (65%). The precipitation selectivity was considered due to the differences in the hydrophobicity of the proteins. The work indicates that by using the AC, RNase A could be efficiently refolded at low urea concentration and purified at high urea concentration from IBs at high protein concentrations. |
| |
| Keywords: | Protein Refolding Bioseparations Aggregation Ribonuclease A Artificial chaperone |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
