참담치(Mytilus coruscus) 유래 항균 펩타이드 분리 및 정제

국문 초록 (Abstract)

항생제 남용의 문제점 및 사용제한에 따른 천연물 유래 항생제 대체제 개발을 위하여 우리나라와 중국, 일본에서만 서식하는 고유종인 참담치의 족부 근육 추출물로부터 항균 펩타이드를 분리 및 정제하였다. C18 역상 컬럼을 사용하여 항균 펩타이드를 정제하였으며, MALDI-TOF/MS로 분석 결과 분자량은 6,701 Da에 해당하였다. edman degradation 방법으로 N-말단 아미노산 서열 분석을 통하여 20개의 아미노산 서열을 밝혔으며 이는 캘리포니아 홍합(Mytilus californianus)의 sperm-specific protein 또는 protamine-like PL-II/PL-IV precursor와 100% 상동성을 지님을 밝혀냈다. 이러한 정보를 바탕으로 ORF를 분석한 결과 60개의 아미노산을 코딩하는 183 bp로 캘리포니아 홍합의 protamine-like PL-II/PL-IV precursor와 아미노산 서열이 100% 일치하였고 유전자 염기서열은 97.2%의 상동성을 나타냈다. 합성된 항균 펩타이드는 그람 양성 균주 Bacillus cereus (MEC, 20.8 μg/ml), Bacillus subtilis(MEC, 0.2 μg/ml), Streptococcus mutans (MEC, 0.2 μg/ml), 그람 음성 균주 Pseudomonas aeruginosa (MEC, 5.7 μg/ml), Escherichia coli (MEC, 2.6 μg/ml) 그리고 진균류 Candida albicans (MEC, 56.3 μg/ml)의 항균활성을 나타냈다. 또한 높은 열안정성을 지녔으며, C. albicans를 제외하고 염안정성을 지닌 항균 펩타이드로 확인되었다. 이를 통해 참담치 유래의 항균 펩타이드는 항생제 대체제의 후보소재로 높은 가능성을 지녔다고 할 수 있다. 따라서 본 연구에서 분리한 참담치 족부 근육 유래 항균 펩타이드는 향후 추가적인 연구를 통하여 항생제를 대체할 수 있는 물질의 개발이 가능할 것으로 사료된다. 그리고 해양무척추동물의 선천성 면역 기작에 대한 정보를 제공할 것으로 기대한다.

다국어 초록 (Multilingual 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.

목차 (Table of Contents)

• 서론
• 재료 및 방법
• 결과 및 고찰
• References
• 초록

참고문헌 (Reference)

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