Chemoenzymatic synthesis of polypeptides in neat 1,1,1,2-tetrafluoroethane solvent

摘要: Chemoenzymatic polypeptide synthesis offers several advantages over chemical or other biological routes, however, the use of aqueous-based media suffers from reverse hydrolysis reactions that challenge peptide chain propagation. Herein, protease subtilisin Carlsberg biocatalyzed poly-L-PheOEt, poly-L-LeuOEt, and copolymers poly-L-PheOEt-co-L-LeuOEt their amino acid ethyl ester substrates in a neat liquid 1,1,1,2-tetrafluoroethane solvent. The products, achieved acceptable yields (ca. 50%), were fully characterized showing relatively high molar mass 20 000 Da for poly-L-PheOEt). This non-toxic low-boiling hydrofluorocarbon enhances enzymatic propagation by limiting owing to its hydrophobic polar characteristics sustain activity solubilize products. Computational molecular dynamic calculations used assess L-PheOEt/L-LeuOEt-solvent polypeptide-solvent interactions this system. Additionally, homopolypeptides displayed higher crystallinity than copolypeptides with random incorporation esters, notwithstanding significantly highest specificity Phe Interestingly, secondary structure characterization products FTIR circular dichroism suggests non-common folding.