2 - 아미노나프탈렌술폰산류의 자외선분광 광도법에 의한 분석

Sohn Joo Hwan(孫周煥),Kim Joo Youn(金宙元) 한국유화학회 1986 한국응용과학기술학회지 Vol.3 No.2

The sulfuric acid sulfonation mixtures of 2-aminonaphthalene, contained 2-aminonaphthalene, Dahl s acid, Broenner s acid, amino-F-acid and Baden acid, can be determined quantitatively by multicomponent spectrophotometric analysis. The analysis was performed in diluted sodium hydroxide aqueous solution and based on the ultraviolet absorption of the sodium salts of each isomers. The determination of quantity of each isomers was performed by subjection the absorbances of the unknown mixture and of its constituents, gathered at a large number of wavelengths, to a least square treatment by an electronic personer computer. This method provided a rapid analysis of such complex mixture, and the standard deviation was ±1.65 mole %.

요추 추간판탈출증 환자의 발병시기에 따른 보존적 치료 결과 비교 연구

윤유석 ( You Suk Youn ),이종수 ( Jong Soo Lee ),하인혁 ( In Hyuk Ha ),김주원 ( Joo Won Kim ),권혁준 ( Hyeok Joon Kwon ) 한방재활의학과학회 2008 한방재활의학과학회지 Vol.18 No.4

Objectives : To investigate therapeutic outcomes of back pain modalities in patients with disc herniation according to the duration by treated with herbal medicine, chuna, acupuncture, bee-venom acupuncture. Methods : We separated 36 patients with disc herniation into three group according to duration; acute patients within 4 week, subacute patients within 5-12 week, and chronic group within 13-24 week. 36 patients with lumbar disc herniation were treated with chuna therapy, acupuncture, bee-venom acupuncture one times a week and took herbal medicine after a meal two times daily. The patients` symptoms were assessed 1 week, 2 week, 4 week, 8 week by Visual analogue scale(VAS), Oswestry disability index(ODI), 36-Item short-form health survey(SF-36). Results : 1. ODI of disability of daily activities showed significantly decreased in acute stage group compared to subacute and chronic stage groups(P<0.05). 2. Physical functioning(PF), bodily pain(BP), social functioning(SF) score of SF-36 showed significantly increased in acute stage group compared to subacute and chronic stage groups(P<0.05). Conclusions : This study suggests that acute stage group compared to subacute and chronic stage groups in patients with lumbar disc herniation is the more effective to improve symptoms treated with herbal medicine, chuna therapy, acupuncture, bee-venom acupuncture.

참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석

오륜경(Ryunkyoung Oh),이민정(Min Jeong Lee),김영옥(Young-Ok Kim),남보혜(Bo-Hye Nam),공희정(Hee Jeong Kong),김주원(Joo-Won Kim),박중연(Jung-Youn Park),서정길(Jung-Kil Seo),김동균(Dong-Gyun Kim) 한국생명과학회 2017 생명과학회지 Vol.27 No.11

참담치 hemocyte에 존재하는 항균 펩타이드를 역상 HPLC column을 사용하여 분리 및 정제하였다. 정제된 펩타이드는 matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) 분석을 통해 분자량이 3330.549 Da이며, edman 분해법을 통해 14개의 N-말단 아미노산 서열을 확보하였다. 분석한 N-말단 서열은 M. californianus의 sperm-specific protein Phi-1과 protamine-like PL-III protein과 각각 93%와 87%의 유사도를 나타냈으며, M. edulis의 sperm-specific protein Phi-1과 87% 일치함을 확인하였다. 또한 openreading frame (ORF)은 306 bp의 길이에 101개의 아미노산을 코딩하고 있음을 밝혔으며, 이는 M. californianus의 sperm-specific protein Phi-1와 93.5% 유사하였다. 분자량과 아미노산 서열에 근거하여 31개 아미노산으로 구성된 펩타이드를 합성하였으며 이는 그람 양성균인 B. subtilis, S. mutans, S. aureus와 그람 음성균인 E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa 그리고 진균류인 C. albicans에 항균 활성을 보였다. 합성한 펩타이드는 항생제 내성균주인 S. aureus CCARM 0203와 S. aureus CCARM 0204에 항균 활성을 보였다. 합성 항균 펩타이드는 넙치 혈장에 대한 용혈현상은 없었고, 세포독성을 확인한 결과 HUVEC cell line에 전혀 독성을 보이지 않았다. 본 연구결과, 참담치의 혈구로부터 분리 및 정제한 sperm-specific protein 유래 항균 펩타이드는 다양한 균주에 항균 활성을 보였고 낮은 세포독성을 가졌으며, 이러한 특성은 본 실험에서 분리한 항균 펩타이드가 항생제 대체재로서 개발 가능성을 제시하고 있다. 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 C18 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.

참담치(Mytilus coruscus) 혈구(hemocyte) 유래 항균 펩타이드 mytilin B의 정제 및 특성 분석

이민정(Min Jeong Lee),오륜경(Ryunkyoung Oh),김영옥(Young-Ok Kim),남보혜(Bo-Hye Nam),공희정(Hee Jeong Kong),김주원(Joo-Won Kim),박중연(Jung Youn Park),서정길(Jung-Kil Seo),김동균(Dong-Gyun Kim) 한국생명과학회 2018 생명과학회지 Vol.28 No.11

참담치(Mytilus coruscus)의 혈구 유래의 항균 펩타이드를 역상 column들을 사용한 reversed-phase high-performance liquid chromatography (RP-HPLC)로 분리 및 정제하였다. 정제된 펩타이드는 matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS)를 통해 분자량이 4041.866 Da으로 밝혀졌으며 Edman degradation법을 통해 25개의 N-말단 서열을 확보하였다. 이는 참담치의 mytilin B precursor 와 100%, mytilin 8 precursor, mytilin 4 precursor와 96% 일치하였다. 또한 103개의 아미노산 서열을 코딩하고 있는 312 bp의 open-reading frame (ORF)을 밝혔으며 이는 참담치의 mytilin B precursor와 100% 일치하였다. 밝혀진 분자량과 아미노산 서열을 바탕으로 C-말단 alanine 잔기의 유무에 따라 2개의 펩타이드를 합성하였으며 이는 mytilin B1과 B2라고 명명하였다. 이들은 그람 양성 균주 Bacillus cereus, Streptococcus parauberis [minimal effective concentrations, MECs 41.6-89.7 μg/ml], 그람 음성 균주 Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, Vibrio ichthyoenteri [MECs 7.4-39.5 μg/ml] 그리고 진균류인 Candida albicans [MECs 26.0-31.8 μg/ml]에 항균활성을 나타냈다. 본 연구 결과, 참담치 혈구유래 mytilin B1과 mytilin B2는 넓은 항균 스펙트럼을 가지고 열과 염분에 대한 안정성이 높으며 용혈현상과 세포독성은 나타나지 않았다. 이러한 특성은 기능성 사료첨가제 및 항생제 대체제로써 충분히 안정적인 역할을 할 뿐만 아니라 추후 mytilin의 구조적 중요성과 참담치의 면역학적 측면에서 다양한 자료를 제시할 것으로 사료된다. We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by C18 reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDITOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs 41.6-89.7 μg/ml), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs 7.4-39.5 μg/ml), and the fungus Candida albicans (MECs 26.0-31.8 μg/ml). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.