Bacterial Cell‐Surface Display of Semisynthetic Cyclic Peptides. (30th October 2018)

• Record Type: Journal Article
• Authors: Palei, Shubhendu; Becher, Kira S.; Nienberg, Christian; Jose, Joachim; Mootz, Henning D.
• Is Part Of: Chembiochem. Volume 20:Number 1(2019)
• Page Start: 72
• Page End: 77
• Publication Date: 2018-10-30
• Subjects: bacterial display -- inteins -- macrocycles -- oxime ligation -- semisynthesis; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201800552
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: Semisynthetic cyclic peptides containing both non‐proteinogenic building blocks, as the synthetic part, and a genetically encoded sequence amenable to DNA‐based randomization hold great potential to expand the chemical space in the quest for novel bioactive peptides. Key to an efficient selection of novel binders to biomacromolecules is a robust method to link their genotype and phenotype. A novel bacterial cell surface display technology has been developed to present cyclic peptides composed of synthetic and genetically encoded fragments in their backbones. The fragments were combined by protein trans ‐splicing and intramolecular oxime ligation. To this end, a split intein half and an unnatural amino acid were displayed with the genetically encoded part on the surface of Escherichia coli . Addition of the synthetic fragment equipped with the split intein partner and an aminooxy moiety, as well as the application of a pH‐shift protocol, resulted in the onsurface formation of the semisynthetic cyclic peptide. This approach will serve for the generation of cyclic peptide libraries suitable for selection by fluorescence‐activated cell sorting, and more generally enables chemical modification of proteins on the bacterial surface. Abstract : Unnatural presentation : Semisynthetic cyclic peptides containing both non‐proteinogenic building blocks and a genetically encoded sequence amenable to DNA‐based randomization hold great potential to expand the chemical space in the quest for novel bioactive peptides. Our approach allows the generation of cyclic peptide libraries on E. coli cells suitable for selection by fluorescence‐activated cells sorting.