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http://dx.doi.org/10.5352/JLS.2017.27.11.1276

The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus  

Oh, Ryunkyoung (Biotechnology Research Division, National Institute of Fisheries Science)
Lee, Min Jeong (Biotechnology Research Division, National Institute of Fisheries Science)
Kim, Young-Ok (Biotechnology Research Division, National Institute of Fisheries Science)
Nam, Bo-Hye (Biotechnology Research Division, National Institute of Fisheries Science)
Kong, Hee Jeong (Biotechnology Research Division, National Institute of Fisheries Science)
Kim, Joo-Won (Biotechnology Research Division, National Institute of Fisheries Science)
Park, Jung-Youn (Biotechnology Research Division, National Institute of Fisheries Science)
Seo, Jung-Kil (Department of Food Science and Biotechnology, Kunsan National University)
Kim, Dong-Gyun (Biotechnology Research Division, National Institute of Fisheries Science)
Publication Information
Journal of Life Science / v.27, no.11, 2017 , pp. 1276-1289 More about this Journal
Abstract
This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.
Keywords
Antimicrobial peptide; Mytilus coruscus; purification; sperm-specific protein;
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Times Cited By KSCI : 2  (Citation Analysis)
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