Preparation of Semisynthetic Peptide Macrocycles Using Split Inteins

  • Shubhendu Palei
  • Henning D. Mootz
Part of the Methods in Molecular Biology book series (MIMB, volume 1495)


Cyclic peptides are highly desired molecules not only for basic research but also for many biomedical and pharmacological applications. Due to their potentially superior physicochemical properties as compared to their linear counterparts, they are considered as ideal candidates for studying protein–protein interactions, among others. Most of the methods developed in recent years to prepare cyclic peptides focus either on a synthetic or a recombinant route. While the former provides access to diversified, noncanonical peptides, including unnatural and d-amino acid, for example, the latter can harness the power of genetic randomization to generate and select from large peptide libraries. Only few approaches have been reported to prepare semisynthetic macrocycles that would benefit from both the advantages associated with synthetic and genetically encoded parts. We describe in this chapter a chemo-enzymatic method to make semisynthetic cyclic peptides in vitro from two fragments using protein trans-splicing and bioorthogonal oxime ligation.


Antibiotics Cyclic peptide Intein Oxime ligation Protein trans-splicing Ssp DnaB intein 



We thank Peter G. Schultz (Scripps Research Institute, La Jolla) for providing the plasmid for AcF incorporation. We acknowledge financial support from the DFG (MO1073/3-2, SPP1623 and Cells in Motion cluster EXC1003) and the International Graduate School of Chemistry in Münster (GSC-MS; Ph.D. stipend to S. P.).


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Institute of Biochemistry, Department of Chemistry and PharmacyInstitute of Biochemistry, University of MuensterMünsterGermany
  2. 2.Institute of Biochemistry, Department of Chemistry and PharmacyUniversity of MuensterMünsterGermany

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