Abstract
Anti-lipopolysaccharide factors (ALFs) with a LPS-binding domain (LBD) are considered to have broad spectrum antimicrobial activities and certain antiviral properties in crustaceans. FcALF2 was one isoform of ALFs isolated from the Chinese shrimp Fenneropenaeus chinensis. Our previous study showed that a modified LBD domain (named LBDv) of FcALF2 exhibited a highly enhanced antimicrobial activity. In the present study, a modified FcALF2 gene (mFcALF2), in which the LBD was substituted by LBDv, was designed and synthesized. This gene was successfully expressed in yeast Pichia pastoris GS115 eukaryotic expression system, and the characteristics of the recombinant protein mFcALF2 were analyzed. mFcALF2 exhibited apparent antibacterial activities against Gram-negative bacteria, including Escherichia coli, Vibrio alginolyticus, Vibrio harveyi, and Vibrio parahaemolyticus, and Gram-positive bacteria, including Bacillus licheniformis and Staphylococcus epidermidis. In addition, mFcALF2 could reduce the propagation of white spot syndrome virus (WSSV) in vivo by pre-incubation with virus. The present study paves the way for developing antimicrobial drugs in aquaculture.
Highlights
Antimicrobial peptides (AMPs), isolated from a variety of different living organisms, have received more and more attention for their contribution to host defense [1,2]
We reversely translated the amino acid sequence into nucleotide sequence, and optimized the codon usage according to the codon bias for the yeast, and synthesized the nucleotide sequences of modified FcALF2 gene (mFcALF2)
The mFcALF2 gene was comprised of 342 bp, with the restriction enzyme sites EcoRI (GAATTC) and Not I (GCGGCCGC) at the opposite ends of the sequence respectively
Summary
Antimicrobial peptides (AMPs), isolated from a variety of different living organisms, have received more and more attention for their contribution to host defense [1,2]. They are considered to be an essential part of the innate immune system since they possess a broad spectrum of antimicrobial activities against bacteria, fungi, some virus, and provide protection against microbial invasion [3,4]. Extensive researches have demonstrated that these AMPs could act as direct antimicrobial agents, and as important regulators of the innate immune system [5,6,7]. AMPs are regarded as potential alternatives to conventional antibiotics since AMPs could hardly lead to bacterial resistance
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