title: Purification and characterization of a novel antibacterial peptide from black soldier fly (Hermetia illucens) larvae.
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.
authors.affiliation: Department of Biological Sciences, Dankook University, Cheonan 330-714, Republic of Korea.
authors.forename: Soon-Ik
authors.initials: SI
authors.lastname: Park
authors.affiliation: 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.
authors.forename: Jong-Wan
authors.initials: JW
authors.lastname: Kim
authors.affiliation: Department of Biological Sciences, Dankook University, Cheonan 330-714, Republic of Korea. Electronic address: smyoe@dankook.ac.kr.
authors.forename: Sung Moon
authors.initials: SM
authors.lastname: Yoe
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D023082:Defensins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004175:Diptera / Q000276:immunology*; D005786:Gene Expression Regulation; D006458:Hemolymph / Q000378:metabolism*; D007113:Immunity, Innate; D019476:Insect Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D055624:Methicillin-Resistant Staphylococcus aureus / Q000276:immunology*; D008969:Molecular Sequence Data; D010691:Phlebotomus / Q000276:immunology; D016415:Sequence Alignment; D013203:Staphylococcal Infections / Q000276:immunology*
chemical_list: D023181:Antimicrobial Cationic Peptides; D023082:Defensins; D019476:Insect Proteins
keywords: Antimicrobial peptide; Black soldier fly; Defensin; Hermetia illucens; Meticillin-resistant Staphylococcus aureus