Journal
BIOMEDICINES
Volume 10, Issue 6, Pages -Publisher
MDPI
DOI: 10.3390/biomedicines10061351
Keywords
antimicrobial peptide; expression; intein; self-cleavage; cecropin-like
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Funding
- Ministry of Science and Technology of Taiwan [110-2113-M-007-021]
- National Taiwan University Hospital Hsinchu Branch
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Cecropins, a type of antimicrobial peptides found in the innate immune system of Cecropia moths, exhibit broad-spectrum antimicrobial and anticancer activities. This study developed a strategy to produce recombinant KR12AGPWR6 with enhanced antimicrobial properties through a self-cleavage system. The results showed that the recombinant KR12AGPWR6 exhibited similar antimicrobial activities compared to the chemically synthesized peptide. This method provides a potential strategy for large-scale production of AMPs.
Cecropins are a family of antimicrobial peptides (AMPs) that are widely found in the innate immune system of Cecropia moths. Cecropins exhibit a broad spectrum of antimicrobial and anticancer activities. The structures of Cecropins are composed of 34-39 amino acids with an N-terminal amphipathic alpha-helix, an AGP hinge and a hydrophobic C-terminal alpha-helix. KR12AGPWR6 was designed based on the Cecropin-like structural feature. In addition to its antimicrobial activities, KR12AGPWR6 also possesses enhanced salt resistance, antiendotoxin and anticancer properties. Herein, we have developed a strategy to produce recombinant KR12AGPWR6 through a salt-sensitive, pH and temperature dependent intein self-cleavage system. The His6-Intein-KR12AGPWR6 was expressed by E. coli and KR12AGPWR6 was released by the self-cleavage of intein under optimized ionic strength, pH and temperature conditions. The molecular weight and structural feature of the recombinant KR12AGPWR6 was determined by MALDI-TOF mass, CD, and NMR spectroscopy. The recombinant KR12AGPWR6 exhibited similar antimicrobial activities compared to the chemically synthesized KR12AGPWR6. Our results provide a potential strategy to obtain large quantities of AMPs and this method is feasible and easy to scale up for commercial production.
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