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Biosynthetic approach for the recombinant expression of cyclotides using E. coli expression systems. Cleavage of the leading signal either in vitro [18] or in vivo [19, 20] by appropriate proteases provides the N-terminal Cys residue required for the cyclization. The backbone cyclization of the linear precursor is then mediated by a modified protein splicing unit or intein. Once the linear precursor is cyclized, folding is spontaneous for kalata B1 and MCoTI-I/II cyclotides [18–20].
