Expression and purification of epinecidin-1 variant (Ac-Var-1) by acid cleavage.
Appl Microbiol Biotechnol
; 108(1): 176, 2024 Jan 26.
Article
em En
| MEDLINE
| ID: mdl-38277014
ABSTRACT
The demand for massive quantities of therapeutic active antimicrobial peptides (AMPs) is high due to their potential as alternatives to antibiotics. However, each antimicrobial peptide has unique properties, necessitating distinct synthesis and purification strategies for their large-scale production. In this study, we bio-synthesized and purified a functional enhanced variant of the AMP epinecidin-1, known as Ac-Var-1 (acid-cleavable variant-1). To generate the active peptide, we cloned the gene for Ac-Var-1 with acid-cleavable site (aspartic acid-proline) into the pET-32a expression vector, purified the fusion protein by His tag enrichment chromatography, and performed acid cleavage to release the active Ac-Var-1 peptide. After acid cleavage, the active Ac-Var-1 was purified and characterized by SDS-PAGE and mass spectrometry. The results from both techniques provided confirmation of the intactness of the purified Ac-Var-1. The Ac-Var-1 inhibited the growth of pathogenic Escherichia coli and Staphylococcus aureus. KEY POINTS ⢠Epinecidin-1 is a well-known antimicrobial peptide having multipotential bioactivities. ⢠Epinecidin-1 variant is developed via the site-directed mutagenesis method to improve its structural stability and bioactivity. ⢠AC-Var-1 development is an economical and easy method to remove peptide from tag protein.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Infecções Estafilocócicas
/
Peptídeos Catiônicos Antimicrobianos
Limite:
Humans
Idioma:
En
Revista:
Appl Microbiol Biotechnol
Ano de publicação:
2024
Tipo de documento:
Article
País de afiliação:
Índia
País de publicação:
Alemanha