In vivo biosynthesis of an Ala-scan library based on the cyclic peptide SFTI-1
- 14 August 2009
- journal article
- research article
- Published by Springer Nature in Amino Acids
- Vol. 38 (5) , 1313-1322
- https://doi.org/10.1007/s00726-009-0338-4
Abstract
We present the in vivo biosynthesis of wild-type sunflower trypsin inhibitor 1 (SFTI-1) inside E. coli cells using an intramolecular native chemical ligation in combination with a modified protein splicing unit. SFTI-1 is a small backbone cyclized polypeptide with a single disulfide bridge. A small library containing multiple Ala mutants was also biosynthesized and its activity was assayed using a trypsin-binding assay. This study clearly demonstrates the exciting possibility of generating large cyclic peptide libraries in live E. coli cells, and is a critical first step for developing in vivo screening and directed evolution technologies using the cyclic peptide SFTI-1 as a molecular scaffold.Keywords
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