Isolation and Purification of Antimicrobial Peptide from Hard-shelled Mussel, Mytilus coruscus

Abstract

In this study, we investigated antimicrobial peptide from the acidified muscle extract of Mytilus coruscus, which mostly inhabits China, Japan, and Korea, to develop a natural product-derived antibiotics substitution in terms of its abuse and restriction. Antimicrobial peptide was purified by C18 reversed-phase high-performance liquid chromatography and was detected as having a molecular mass of 6,701 Da by MALDI-TOF/MS. The N-terminal amino acid sequence of the purified peak was obtained from edman degradation, and 20 identified residues shown 100% identity with the N-terminus region of sperm-specific protein and protamine-like PL-II/PL-IV precursor of Mytilus californianus. We also identified 60 open-reading frame (ORF) encoding amino acids with 183 bp of purified peptide based on the obtained amino acid residues. The amino acid sequence of ORF showed 100% and the nucleotide sequence revealed 97.2% identity with the protamine-like PL-II/PL-IV precursor of Mytilus californianus. Synthesized antimicrobial peptide showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus (minimal effective concentration [MEC], 20.8 μg/ml), Bacillus subtilis (MEC, 0.2 μg/ ml), Streptococcus mutans (MEC, 0.2 μg/ml), gram-negative bacteria including Pseudomonas aeruginosa (MEC, 5.7 μg/ml), Escherichia coli (MEC, 2.6 μg/ml) and fungi, Candida albicans (MEC, 56.3 μg/ml). In addition, synthesized peptide showed stable activities under heat and salt conditions against gram-positive and gram-negative bacteria, but was inhibited by salt against only C. albicans. With these results, isolated peptide from M. coruscus could be an alternative agent to antibiotics for defending against pathogenic microorganisms, and helpful information to understand the innate immune system of marine invertebrates.Key words : Antimicrobial peptide, hard shelled mussle, Mytilus coruscus, purification *Corresponding author *Tel : +82-51-720-2454, Fax : +82-51-720-2456 *E-mail : combikola@korea.kr This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Therapeutic Potential of Marine Bioactive Peptides against Human Immunodeficiency Virus: Recent Evidence, Challenges, and Future Trends

“…The cyclic and linear peptides derived from marine ascidians have been found with anti-HIV potential with many other biological properties. A cyclic heptapeptide, mollamide B (Figure 5), isolated from the colonial tunicate Didemnum molle indicated cytotoxicity against HIV-1 in human PBMCs at an EC50 value of 48.7 μM [142]. Two peptides, P1 and P2 [sequences LLEYSI (P1), and LLEYSL (P2)] have been isolated from the oyster (Crassostrea gigas) proteins hydrolysate prepared with thermolysin and have exhibited competitive HIV-1 protease inhibition activity (IC50 values of 20 and 15 nM, respectively, and Ki values of 13 and 10 nM, respectively) [143][144][145].…”

“…The cyclic and linear peptides derived from marine ascidians have been found with anti-HIV potential with many other biological properties. A cyclic heptapeptide, mollamide B ( Figure 5 ), isolated from the colonial tunicate Didemnum molle indicated cytotoxicity against HIV-1 in human PBMCs at an EC 50 value of 48.7 μM [ 142 ].…”