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Production of an Antimicrobial Peptide AN 5‐1 in Escherichia coli and its Dual Mechanisms Against Bacteria
Author(s) -
Yi Tonghui,
Huang Yibing,
Chen Yuxin
Publication year - 2015
Publication title -
chemical biology and drug design
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.59
H-Index - 77
eISSN - 1747-0285
pISSN - 1747-0277
DOI - 10.1111/cbdd.12449
Subject(s) - bacteria , escherichia coli , antimicrobial , antimicrobial peptides , peptide , biology , bacterial cell structure , biochemistry , microbiology and biotechnology , intracellular , chemistry , gene , genetics
AN 5‐1 ( YSKSLPLSVLNP ) is an antimicrobial peptide isolated from the fermentation broth of Paenibacillus alvei strain AN 5 ( J Ind Microb Biotechnol 2013; 40 : 571–9). In this study, we report the application of ubiquitin fusion technology to the expression and purification of AN 5‐1. Minimum inhibitory concentration ( MIC ) and measurement of hemolytic activity ( MHC ) were measured to confirm the biological activities of the expressed AN 5‐1. Bacterial cell membrane permeabilization was investigated to show the interaction between the AN 5‐1 and the bacterial cytoplasmic membrane. Furthermore, intracellular activities of the AN 5‐1 were determined by genomic DNA interaction assays. The results revealed AN 5‐1 damaging bacterial membranes and binding to bacterial genomic DNA to inhibit cellular functions, suggesting that it has multiple intracellular targets in bacteria. The application of ubiquitin fusion technology may be an excellent approach for industrial production to the expression and purification of antimicrobial peptide. Furthermore, AN 5‐1 was demonstrated as an antimicrobial peptide with great potentials, as bacterial resistance to AN 5‐1 would be not expected, due to the dual mechanisms of AN 5‐1 against bacteria.