Peptide synthesis in neat organic solvents with novel thermostable proteases |
| |
Institution: | 1. Department of Cardiology, Academic Medical Center, Amsterdam, The Netherlands;2. Interuniversity Cardiology Institute of the Netherlands, Utrecht, The Netherlands;3. Haga Hospital, The Hague, The Netherlands;4. Department of Cardiology, Leiden Medical Center, Leiden, The Netherlands;5. Department of Vascular Medicine, Academic Medical Center, Amsterdam, The Netherlands;6. Department of Cardiology, University Medical Center Groningen, The Netherlands;7. Department of Cardiology, Radboud University Medical Center, The Netherlands;8. Department of Cardiology, University Medical Center Utrecht, The Netherlands;9. Department of Cardiology, VU University Medical Center, Medical Center, The Netherlands;10. Department of Vascular Medicine, Academic Medical Center Amsterdam, The Netherlands;11. Department of Clinical Epidemiology and Biostatistics, Academic Medical Center, Amsterdam, The Netherlands |
| |
Abstract: | Biocatalytic peptide synthesis will benefit from enzymes that are active at low water levels in organic solvent compositions that allow good substrate and product solubility. To explore the use of proteases from thermophiles for peptide synthesis under such conditions, putative protease genes of the subtilase class were cloned from Thermus aquaticus and Deinococcus geothermalis and expressed in Escherichia coli. The purified enzymes were highly thermostable and catalyzed efficient peptide bond synthesis at 80 °C and 60 °C in neat acetonitrile with excellent conversion (>90%). The enzymes tolerated high levels of N,N-dimethylformamide (DMF) as a cosolvent (40–50% v/v), which improved substrate solubility and gave good conversion in 5+3 peptide condensation reactions. The results suggest that proteases from thermophiles can be used for peptide synthesis under harsh reaction conditions. |
| |
Keywords: | Thermostable Thermophile Protease Subtilase Organic solvent Peptide synthesis |
本文献已被 ScienceDirect 等数据库收录! |
|