High Level Expression and Purification of Cecropin-like Antimicrobial Peptides in .
Journal
Biomedicines
Journal Volume
10
Journal Issue
6
ISSN
2227-9059
Date Issued
2022-06-08
Author(s)
Wu, Chih-Lung
Chih, Ya-Han
Hsieh, Hsin-Ying
Peng, Kuang-Li
Lee, Yi-Zong
Sue, Shih-Che
Cheng, Jya-Wei
Abstract
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 α-helix, an AGP hinge and a hydrophobic C-terminal α-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 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.
Subjects
antimicrobial peptide
cecropin-like
expression
intein
self-cleavage
SDGs
Type
journal article