피부 병원균에 대한 톱니모자반 추출물의 항균 시너지 효과

김윤혜(Yun Hye Kim),김지훈(Ji-Hoon Kim),김덕훈(Deok-Hoon Kim),김송희(Song-Hee Kim),김형락(Hyeung-Rak Kim),김영목(Young-Mog Kim) 한국식품과학회 2016 한국식품과학회지 Vol.48 No.3

S. aureus, S. epidermidis, P. aeruginosa, P. acnes와 C. albicans는 사람의 피부에서 발생하는 질병과 밀접한 관련을 가지는 대표적인 병원성 미생물로 알려져 있다. 본 연구에서는 해조류 중에서도 항균 활성에 대한 연구가 미미한 모자반을 대상으로 피부 병원균에 대한 항균 효과를 조사하였다. 국내의 연해에 자생하는 7종의 모자반 추출물 중에서 disc diffusion assay와 MIC assay를 통해 가장 뛰어난 항균 효과를 나타낸 톱니모자반을 후속 연구를 위한 후보 물질로 선정하고 연구를 진행하였다. 톱니모자반 추출물의 유기용매 분획층 중에서, 노말-헥세인 분획층이 S. aureus, S. epidermidis, P. aeruginosa, P. acnes 및 C. albicans에 대한 MIC값이 32-256 μg/mL로 가장 뛰어난 항균 활성을 나타내었다. 이에 피부 병원균들에 대한 치료제로 사용되고 있지만 내성균의 출현으로 효능이 거의 없는 항생제들인 테트라사이클린, 에리트로마이신, 린코마이신과 플루코나졸과 항균 활성이 뛰어난 것으로 나타난 톱니모자반의 노말-헥세인 분획층과의 병용 사용에 의한 항균 시너지 효과를 조사하였다. 그 결과, 톱니 모자반 노말-헥세인 분획층과 이들 항생제와의 병용 사용에 의해 피부 병원균에 대한 항생제와 톱니모자반 노말-헥세인 분획층의 MIC값이 4-32배 감소되었고, 톱니모자반 노말-헥세인 분획층과 이들 항생제와의 병용 시 median FIC값이 0.26-0.55로 항균 시너지 효과를 나타내었다. 즉, 항생제와 톱니모자반 노말-헥세인 분획층과의 병용 사용은 피부 병원균에 대한 이들 항생제의 감수성을 회복시키는데 크게 기여하는 것으로 나타났다. The object of this study was to develop an alternative way to treat human skin pathogens using marine algae. During this study, we observed that the ethanolic extract of the edible brown algae [Sargassum serratifolium (C. Agardh) C. Agardh] exhibited potential antimicrobial activity against pathogenic commensal bacteria related with acne vulgaris (Propionibacterium acnes, Staphylococcus epidermidis, Staphylococcus aureus and Pseudomonas aeruginosa), and Candida albicans which causes cutaneous candidiasis. Among the solvent-soluble fractions from the ethanolic extract, a hexane-soluble fraction showed the strongest antimicrobial activity against all tested human skin pathogens with MIC values ranging from 32 to 512 μg/mL. In addition, the hexane fraction exhibited a synergistic antimicrobial activity with commercial antibiotics used in the treatment of acne vulgaris or cutaneous candidiasis. Thus, this study suggests that S. serratifolium extract could be a potential source of natural antimicrobial agents or a pharmaceutical component against human skin pathogens.

LPS로 유도된 RAW 264.7 대식세포에 대한 애기외톨개 모자반 (Myagropsis yendoi) 에틸아세테이트 분획물의 항염증 효과

김보운 ( Bowoon Kim ),김재일 ( Jae Il Kim ),김형락 ( Hyeung Rak Kim ),변대석 ( Dae Seok Byun ) 한국수산과학회 2014 한국수산과학회지 Vol.47 No.5

An ethanolic extract from Myagropsis yendoi was fractionated using several solvents. Among these, an ethyl acetate fraction (Myagropsis yendoi ethyl acetate fraction: MYE) showed the highest anti-inflammatory activity based on inhibition of lipopolysaccharides (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells. We thus investi-gated the molecular mechanisms underlying MYE’s inhibitory effects. Pretreatment of cells with up to 30 μg/mL of MYE significantly inhibited NO production and inducible nitric oxide synthase expression in a dose-dependent man-ner (P<0.05). Similarly, MYE markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α, as well as their mRNA levels. While the nuclear translocation of nuclear factor-kappa B (NF-κB) was strongly suppressed by MYE, the activation of a nuclear factor erythroid 2-re-lated factor (Nrf2) was increased. Moreover, MYE significantly reduced the phosphorylation of JNK, p38 MAPK, and phosphatidylinositol 3-kinase/Akt in LPS-stimulated cells. These results indicate that MYE contains anti-inflam-matory compounds, and that it might be used as a dietary supplement for the prevention of inflammatory diseases.

멸치 육과 내장으로부터 분리한 Cathepsin L, Chymotrypsin 및 Trypsin의 단백질분해 특성

번재형 ( Jae Hyeung Pyeun ),허민수 ( Min Soo Heu ),조득문 ( Deuk Moon Cho ),김형락 ( Hyeung Rak Kim ) 한국수산학회 1995 한국수산과학회지 Vol.28 No.5

어류의 사후 초기의 변화를 육 및 장기조직중에 분포하는 단백질분해효소의 작용과 관련하여 검토할 목적으로 멸치의 육 및 장기에서 분리한 cathepsin L과 chymotrypsin 및 trypsin의 단백질 기질에 대한 특성과 근원섬유단백질에 대한 분해능을 전기영동적으로 분석하여 다음의 결론을 얻었다. 이들 세 효소의 casein에 대한 친화도는 유사하였고, 근원섬유단백질에 대한 친화도는 casein에 대한 친화도보다 높았다. 멸치와 방어의 근원섬유단백질에 대한 cathepsin L과 chymotrypsin의 활성은 trypsin보다 훨씬 높게 나타났다. 0~25%까지의 식염농도에서 세 효소의 단백질분해활성은 식염의 농도에 반비례하였으며, 식염의 공존상태에서 세 효소는 casein 보다 근원섬유단백질에 대하여 높은 활성을 나타내었다. 근원섬유단백질의 효소 분해시에 cathepsin L은 chymotrypsin과 trypsin에 비하여 염농도와 온도에 의한 영향이 적었다. 따라서, 멸치의 사후변화와 젓갈 숙성중의 자가소화는 trypsin보다는 cathepsin L과 chymotrypsin의 단백질분해활성이 더욱 깊이 관여할 것으로 판단된다. Proteolytic properties of enzymes from the muscle and viscera of anchovy have been examined. Cathepsin L, chymotrypsin, and trypsin showed similar Km values for casein. However, they had higher Km values for myofibrillar proteins than those for casein. The k(cat) of cathepsin L and chymotrypsin for myofibrillar proteins were higher than that of trypsin, and also cathepsin L and chymotrypsin caused higher hydrolysis in myofibrillar proteins of anchovy and yellowtail. In the presence of sodium chloride (0~25%), proteolytic activity for myofibrillar proteins from yellowtail was higher than that for casein. Proteolytic activity was decreased with the increase of sodium chloride concentration. Cathepsin L had been less affected by NaCl concentration and temperature on the hydrolysis of myofibrillar proteins than chymotrypsin and trypsin. Cathepsin L and chymotrypsin were more responsible to the autolysis of muscle proteins from fish than trypsin.

LPS로 유도된 RAW 264.7 대식세포에 대한 대황(Eisenia bicyclis) 헥산 분획물의 항염증 효과

김보운 ( Bowoon Kim ),최창근 ( Chang-geun Choi ),김재일 ( Jae-il Kim ),김형락 ( Hyeung-rak Kim ) 한국수산과학회(구 한국수산학회) 2021 한국수산과학회지 Vol.54 No.2

Eisenia bicyclis is known to have secondary metabolites exhibiting various biological activities. In a preliminary study, the n-hexane fraction obtained from the ethanolic extract of E. bicyclis showed higher anti-inflammatory activity than the ethyl acetate and butyl alcohol fractions based on the inhibition of lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production in RAW 264.7 cells. Using this fraction (E. bicyclis hexane fraction, EHF), we investigated the molecular mechanisms underlying its anti-inflammatory effect in LPS-stimulated RAW 264.7 cells. Pretreatment of the cells with up to 50 μg/mL EHF significantly inhibited NO and prostaglandin E2 production as well as their responsible enzyme proteins and mRNAs, in a dose-dependent manner (P<0.05). Similarly, EHF markedly reduced the production of pro-inflammatory cytokines, such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α as well as their mRNA levels. Nuclear translocation of nuclear factor-kappa B (NF-κB) was strongly suppressed by EHF treatment. EHF significantly reduced the phosphorylation of mitogen-activated protein kinases and phosphatidylinositol 3-kinase/Akt in LPS-stimulated cells. Moreover, EHF reduced ear edema in phorbol myristate acetate (PMA)-induced mice. These results indicate that EHF contains potent anti-inflammatory compounds, which may be used as a dietary supplement for the prevention of inflammatory diseases.

식용 갈조류의 알긴산 조성

이동수(Dong Soo Lee),김형락(Hyeung Rak Kim),조득문(Deuk Moon Cho),남택정(Taek Jeong Nam),변재형(Jae Hyeung Pyeun) 한국수산과학회 1998 한국수산과학회지 Vol.31 No.1

Uronate compositions and molecular weights of alginates from the four kinds of brown algae, sea mustard(Undaria pinnatifida), sea tangle(Laminaria japonica), gulf weed(Sargassum fulvellum), and seaweed fusiforme(Hizikia fusiforme), in regard with the harvesting season were investigated. Sea mustard contained the highest amount of alginates in the four kinds of brown algae. _D-Mannuronic acid to _L-guluronic acid(M/G) ratio of the alginates was high in order of seaweed fusiforme, gulf weed, sea mustard, and sea tangle, and especially in water-soluble alginate. Molecular weights of the alginates were greater with the growing period ranging in 4,500-4,800kDa for sea tangle, 4,000-4,200kDa for sea mustard, 3,300-3,400kDa for seaweed fusiforme, and 3,000-3,200kDa for gulfweed. In water-soluble alginate of sea mustard, M/G ratio was much higher in sporophyll than in midrib and blade.

식용해조류중의 미량요소와 특수기능성 당질: 2. 산지와 채취시기별 식이성섬유질 함량의 변화와 해조다당류의 분포

김두상 ( Doo Sang Kim ),이동수 ( Dong Soo Lee ),조득문 ( Deuk Moon Cho ),김형락 ( Hyeung Rak Kim ),변재형 ( Jae Hyeung Pyeun ) 한국수산학회 1995 한국수산과학회지 Vol.28 No.3

우리 나라 대표적인 식용해조 (녹조류-참홑파래, 청각; 갈조류-미역, 톳, 모자반, 다시마; 홍조류-얼룩도박, 참김, 불등풀가사리) 9종을 대상으로 가용성 식이섬유질, 불용성 식이섬유질 및 총 식이섬유질의 함량을 주요 산지 (기장, 충무, 여수)와 채취 시기별로 측정·검토하고, 특히 갈조류와 홍조류에 대하여는 해조 다당류의 분포를 분석 비교하였다. 총 식이섬유질은 녹조류 중의 참홑파래가 25.4~38.1% (이하 건물중의 비율), 청각은 35.4~43.8%를, 갈조류 중의 미역은 34.2~48.8%, 톳은 37.5~47.8%, 모자반은 42.9~71.3%, 다시마는 37.1~45.1%를, 그리고, 홍조류 중의 참김은 31.3~40.5%, 얼룩도박은 51.5~60.4%, 불등풀가사리는 57.1~65.8%를 각각 함유하였다. 가용성 식이섬유질과 불용성 식이섬유질을 공통적으로 많이 함유한 해조류는 각각 불등풀가사리와 모자반이었다. 총 식이섬유질에 대한 가용성 식이섬유질의 함유 비율은 녹조류 중에서는 참홑파래 (43.7~64.8%), 갈조류 중에서는 미역 (17.5~31.3%), 그리고 홍조류 중에서는 불등풀가사리 (44.7~63.2%)가 높은 값을 보였다. 해조 다당류의 분포상의 특정을 보면, 갈조류는 알칼리 가용성 알긴산을 가장 많이 함유하였으며 (9.0~15.1%), 홍조류 중의 참김은 폴피란 (5.8%)을, 그리고 불등풀가사리와 얼룩도박은 한천 (20.0%)과 카라기난 (23.8%)을 각각 많이 함유하는 차이를 보였다. This report described on the soluble, insoluble, and total dietary fiber level depending on the harvesting seasons and habitats in 9 species of marine algae. Total dietary fiber contents were comprised 25.4~38.1% (dry basis) in green laver and 35.4~43.8% in sea staghorn of green algae, 34.2~48.8% in sea mustard, 37.5~47.8% in seaweed fusiforme, 42.9­~71.3% in gulf weed, and 37.1~45.1% in sea tangle of brown algae, and 31.3~40.5% in laver, 51.5~60.4% in seaweed dilatata, and 57.1~65.8% in seaweed furcata of red algae. Relatively high levels of both soluble and insoluble dietary fibers were found in seaweed furcata and gulf weed. The ratio of soluble dietary fiber to total dietary fiber was the highest in green laver (43.7~­64.8%), sea mustard (17.5~31.3%), and seaweed furcata (44.7~63.2%) in their respective groups. The highest level of algal polysaccharides was confirmed to be an alkali-soluble alginic acid (9.0~15.1%) in whole brown algae, porphyran (5.8%) in laver, agar (20.0%) in seaweed furcata, and carrageenan (23.8%) in seaweed dilatata of red algae.

발효 , 세포배양 , 생물공정 단백질 분해효소 생산을 위한 Aspergillus oryzae PF 균주의 배지조성

김두상(Doo Sang Kim),김형락(Hyeung Rak Kim),남택정(Taek Jeong Nam),변재형(Jae Hyeung Pyeun) 한국미생물생명공학회 1999 한국미생물·생명공학회지 Vol.27 No.5

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PCR-DGGE를 이용한 해양미생물의 다양성 조사

김영진 ( Yeong Jin Kim ),조효진 ( Hyo Jin Cho ),유선녕 ( Sun Nyoung Yu ),김광연 ( Kwang Youn Kim ),김형락 ( Hyeung Rak Kim ),안순철 ( Soon Cheol Ahn ) 한국수산과학회 2007 한국수산과학회지 Vol.40 No.6

국내산 우뭇가사리로부터 미생물 배지용 한천의 pilot 규모 정제와 특성

김두상(Doo Sang Kim),김형락(Hyeung Rak Kim),김정한(Jeong Han Kim),변재형(Jae Hyeong Pyeun) 한국수산과학회 2000 한국수산과학회지 Vol.33 No.1

Agar for microbiological medium was prepared with pilot-scale for industrial application by the process of microfiltration (0.4 ㎛ pore size) in 40∼50℃, washing with 50℃ water, and treatment with 0.25 N NaOH at 70℃. Transparency, gel strength, viscosity, sulfate content, and syneresis ratio of agar prepared from Gelidium amansii was compared with commercial agar for microbiological medium. Gel strength and transparency were increased with processing, however, it`s viscosity, sulfate content, and syneresis ratio were reduced. The final agar product was superior to commercial agar for microbiological medium.

톱니모자반(Sargassum serratifolium) 추출물의 항산화 활성 증가를 위한 추출공정 최적화

김성희 ( Seong-hee Kim ),고지윤 ( Ji-yun Ko ),김형락 ( Hyeung-rak Kim ),이양봉 ( Yang-bong Lee ) 한국수산과학회(구 한국수산학회) 2021 한국수산과학회지 Vol.54 No.1

Sargassum serratifolium is a rich source of antioxidant meroterpenoids. The optimal extraction temperature and time for obtaining maximal antioxidant yield, antioxidant activity, and phenolic content from Sargassum serratifolium were determined using response surface methodology (RSM). The ranges of the independent variables for extraction temperature and time were 30-70°C and 12-36 h, respectively. With increasing temperature and time, the yield increased significantly, while DPPH (2,2-dipheny-1-picrylhydrazyl) radical-scavenging activity and total phenolic content decreased significantly. The optimal extraction temperature and time obtained by RSM were 54°C and 7 h, respectively, providing a yield of 8.2%, DPPH radical-scavenging activity of 60%, and total phenolic content of 163 mg GAE/g. The findings of this study provide useful information for the development of Sargassum serratifolium extraction processes for the food and cosmetic industries.