Recombinant Production of The Cyclotide Kalata B1 by Conditional Split Inteins

This study describes the design, production, and characterization of a novel conditional intein system for the recombinant production of cyclic peptides. The system is based on two key features: (1) a promiscuous extein recognition site allowing cyclization of virtually any peptide, and (2) a secondary split site within the intein itself enabling triggered splicing at will. Two intein precursors were recombinantly expressed, purified, and then self‐assembled in vitro to cyclize the model peptide kalata B1 (kB1). Cyclized kB1 was successfully purified, refolded, and characterized by mass spectrometry and NMR, demonstrating correct disulfide bond formation and identical structure to synthetic kB1. Importantly, the intein‐derived kB1 retained full biological activity as evidenced by insect cell toxicity assays. This work establishes a versatile and efficient approach for intein‐mediated protein cyclization with potential applications in bioengineering and peptide discovery.

• Publication year

  2024

• Venue

  ChemBioChem

• Publication date

  2024-09-06

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1002/cbic.202400591PMID 39239927
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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