Related Experiment Videos
Thiolsubtilisin as an instrument for peptide synthesis. Preparation and properties
S V Kolobanova1, E N Lysogorskaya, Filippova IYu
1School of Chemistry, Lomonosov Moscow State University, Russia.
Biochemistry. Biokhimiia
|March 1, 1997
Abstract:
A convenient procedure for thiolsubtilisin purification from an admixture of subtilisin involving affinity chromatography on bacitracin-Sepharose is presented. Thiolsubtilisin activity was measured by hydrolysis of p-nitrophenyl acetate, p-nitroanilide-peptide (Glp-Ala-Ala-Leu-pNA), and azocasein. The thiolenzyme catalyzes peptide synthesis. Under these conditions only activated peptide esters, e.g., p-chlorophenyl, N-hydroxysuccinimide, or p-nitrophenyl esters form peptide bonds during interaction with appropriate nucleophiles such as peptides and their derivatives and amino acid amides.