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Dev Comp Immunol. 2015 Sep;52(1):98-106. doi: 10.1016/j.dci.2015.04.018. Epub 2015 May 5.

Purification and characterization of a novel antibacterial peptide from black soldier fly (Hermetia illucens) larvae.

Author information

1
Department of Biological Sciences, Dankook University, Cheonan 330-714, Republic of Korea.
2
Department of Nanobiomedical Science, Dankook University Graduate School, Cheonan 330-714, Republic of Korea; Institute of Tissue Regeneration Engineering, Dankook University, Cheonan 330-714, Republic of Korea.
3
Department of Biological Sciences, Dankook University, Cheonan 330-714, Republic of Korea. Electronic address: smyoe@dankook.ac.kr.

Abstract

In this study, we induced and purified a novel antimicrobial peptide exhibiting activity against Gram-positive bacteria from the immunized hemolymph of Hermetia illucens larvae. The immunized hemolymph was extracted, and the novel defensin-like peptide 4 (DLP4) was purified using solid-phase extraction and reverse-phase chromatography. The purified DLP4 demonstrated a molecular weight of 4267 Da, as determined using the matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF) method. From analysis of DLP4 by N-terminal amino acid sequencing using Edman degradation, combined with MALDI-TOF and rapid amplification of cDNA ends-polymerase chain reaction (RACE-PCR), the amino acid sequence of the mature peptide was determined to be ATCDLLSPFKVGHAACAAHCIARGKRGGWCDKRAVCNCRK. In NCBI BLAST, the amino acid sequence of DPL4 was found to be 75% identical to the Phlebotomus duboscqi defensin. Analysis of the minimal inhibitory concentration (MIC) revealed that DLP4 have antibacterial effects against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA). The expression of DLP4 transcripts in several tissues after bacterial challenge was measured by quantitative real-time PCR. Expression of the DLP4 gene hardly occurred throughout the body before immunization, but was mostly evident in the fat body after immunization.

KEYWORDS:

Antimicrobial peptide; Black soldier fly; Defensin; Hermetia illucens; Meticillin-resistant Staphylococcus aureus

PMID:
25956195
DOI:
10.1016/j.dci.2015.04.018
[Indexed for MEDLINE]

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