High-level Expression of Antimicrobial Peptide Mytimacin-4 in Escherichia coli and Identification of Its Activity
ZHAO Nannan
HUANG Jiaping
CUI Jinming
JIA Aiqing
ZHU Xuan
LIN Ying
WANG Jian
Abstract：Mytimacin-4 from Mytilus galloprovincialis has strong bacteriostatic activity against gram-positive and gram-negative bacteria.However,the production of mytimacin-4 has been rarely studied,which limits its subsequent product development and application.In this study,the gene sequence of mytimacin-4 was codon optimized in Escherichia coli and constructed in the expression vector pET-28a.Then,pET-28a-mytimacin-4 and each of the expression vectors containing different combinations of molecular chaperones(pG-KJE8,pKJE7,pGro7,pG-Tf2,and pTf16)were co-transformed into E.coli BL21(DE3)for induced expression and activity identification.The results showed that the co-expression of molecular chaperones significantly improved the soluble expression of mytimacin-4 in Escherichia coli.Notably,the optimized yields of the recombinant strain BL21(DE3)/pET-28a-mn4/pG-KJE8 ranged from 200 to 400 mg/L,the highest reported.Purified mytimacin-4 was obtained by metal-ion affinity chromatography product of the expression products.The purified mytimacin-4 showed strong inhibitory activity against Staphylococcus aureus,E.coli,and Vibrio parahaemolyticus.In this study,an engineering strain with high expression of mytimacin-4 was constructed,which provided technical support for applying mytimacin-4 in veterinary medicine and aquatic feed.
Keywords：antimicrobial peptide mytimacin-4molecular chaperonebacteriostatic activity
Publication Date：2025-05-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：10( 114-123 )