Salt-sensitive intein for large-scale polypeptide production

High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in Escherichia coli

“…Intein is a protein segment that can cleave itself from a whole protein sequence and ligate the remaining N-terminal and C-terminal portions (the exteins) with a peptide bond [ 21 ]. Until now, over 600 inteins with different lengths have been identified.…”

“…The specific cleavage-ligation function of intein has enabled various applications, such as protein engineering, isotope labeling, biomaterials, cyclization, and protein purification [ 54 , 55 ]. Recently, we have created an N-terminal mutated intein that has no N-terminal cleavage activity but preserves its C-terminal cleavage activity [ 21 ]. We have shown that this mutated intein can be used as a fusion tag and can self-cleave from the target protein.…”

“…We have shown that this mutated intein can be used as a fusion tag and can self-cleave from the target protein. Moreover, we have shown that the efficiency of the intein self-cleavage is dependent on ionic strength, pH, temperature, and reaction time [ 21 ].…”

“…There are two methods to produce AMPs, chemical synthesis and biosynthesis [ 21 ]. Chemical synthesis, such as solid-phase synthesis can provide efficiency and flexibility in the development of AMPs at a laboratory scale [ 22 ].…”

“…On the other hand, chemical cleavage by CNBr or hydroxylamine has disadvantages, such as hazardousness and side-chain modifications [ 30 , 31 , 32 ]. Recently, we developed a salt-sensitive and self-cleavage Nostoc punctiforme ( Npu ) DnaE intein expression system [ 21 ]. The self-cleavage can be controlled by ionic strength, pH, and temperature.…”