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Korean J. Microbiol. Biotechnol. Vol. 40, No. 2, 144 151 (2012) http://dx.doi.org/10.4014/kjmb.1203.03005 pissn 1598-642X eissn 2234-7305 35 종해조류추출물의병원성세균및 Candida sp. 진균에대한항균활성평가 김미선 권경진 이민진 안선미 손호용 * 안동대학교식품영양학과 Received : March 14, 2012 / Revised : April 9, 2012 / Accepted : April 17, 2012 Evaluation of the Antimicrobial Activities of 35 Seaweed Extracts against Pathogenic Bacteria and Candida sp.. Kim, Mi-Sun, Kyung-Jin Kwon, Min-Jin Lee, Seon-Mi Ahn, and Ho-Yong Sohn*. Department of Food and Nutrition, Andong National University, Andong 760-749, Korea In the course of this study aimed at the development of functional food ingredients from seaweeds, the in vitro antimicrobial activities of methanol extracts prepared from 35 different seaweeds (17 phaeophyta, 11 rhodophyta and 7 chlorophyta) were determined against food-borne diseases and pathogenic microorganisms including multi-drug resistant (MDR) Pseudomonas sp. and Candida sp. Based on disc-diffusion assays at 500 g/disc concentration of the methanol extracts, Ishige okamurai, I. foliacea, Sargassum confusum, and S. yamade exhibited strong antibacterial activities in a broad-spectrum, except against Pseudomonas aeruginosa. In addition to the latter four seaweeds, Ecklonia stolonifera, E. cava and Eisenia bicyclis also demonstrated antifungal activity against C. albicans. Among these 8 selected seaweeds, I. okamurai, I. foliacea, and S. yamade exhibited strong hemolytic activity (55-93%) at 500 g/ml against human RBC. Organic solvent sequential fractions using hexane, ethylacetate and butanol, and water residues were prepared from the 8 selected seaweeds and their anti-candida sp. activities were further determined. The ethylacetate and butanol fraction of I. okamurai, and the hexane fraction of I. foliacea demonstrated antifungal activity against MDR-pathogenic Candida sp. Although the solvent fractions had no activity against MDR-Pseudomonas sp., our results suggest that seaweeds, especially Ishige okamurai, I. foliacea, S. confusum, and S. yamade could be developed as broad-spectrum antimicrobial ingredients. Keywords: Seaweeds, antimicrobial activity, multi-drug resistance, pathogenic candida sp, ishige okamurai, ishige foliacea 서 인류는건강한삶과식량확보를위해미생물에의한인체감염및식품오염에끊임없이대항해왔다. 실제생명공학과정밀화학의발전으로인해다양한의약용, 식품용, 농업용화학항균제가개발되어현대사회산업전분야에광범위하게이용되고있다. 그러나미생물들은고빈도사용항균제에대해스스로내성을획득하고, 나아가유사구조의항균제들에대해교차내성을나타내므로 [11, 25], 신규항균제개발은지속적으로이루어져야만한다. 또한지구온난화에따라미생물성식중독및병원미생물감염위험성이증대되고있으며, 식품원재료의대량재배, 유통및저장시스템에서의미생물제어어려움을고려할때식품에직접이용가능 론 *Corresponding author Tel: +82-54-820-5491, Fax: +82-54-820-7804 E-mail: hysohn@andong.ac.kr 한항균제개발이필수적이며, 따라서안전성이확보된천연물유래의항균제개발이절실한상태이다 [26-28]. 현재까지개발된대부분의생물소재유래항균제는토양방선균, 식물및곤충을포함한육상생물기원이었으나 [15, 26-28], 최근에는해양생물자원으로부터신규항균제를개발하려는시도가활발히진행되고있다 [2, 13, 18-21]. 이는지구표면적의 70% 를차지하는해양에는지구생물종의 80% 이상의다양한생물종이서식하고있으며, 다양한해양생물들은육상생물과는전혀다른생활환경으로인해특이한생리활성물질을보유하고있기때문이다 [4,16]. 따라서광범위한해양생물자원의유용생리활성평가에따른생물소재개발연구가지속적으로필요하며, 특히한국의경우세계 4 위의생산량을가지는해조류 [4] 에대한보다적극적인생리활성연구및산업적이용연구가필요하다. 해조류의경우 fucoidans 와같은다당류의항혈전 [22], 항산화 [2, 12, 23], 항암 [3], nitrite 및중금속제거활성 [6, 24], 고지혈증및고혈압예방 [8, 17], 항당뇨 [16], 항돌연변이 [14],

BIOACTIVITY OF SEAWEEDS 145 멜라린생성억제 [5, 9], elastase 저해 [5] 및항염증활성 [9] 등다양한유용활성이보고되어있으며, 최근에는해조류의항균활성에대한연구가증가되고있다. 현재까지해조류의항균활성은서실, 모로우붉은실, 다시마, 갈파래, 청각, 감태, 곰피, 미역, 패, 넓패, 몽우리청각등각각의해조류를대상으로어병세균, 항충치균, 황색포도상구균및대장균에한정하여보고한바있다 [2, 7, 10, 13, 14, 21]. 그러나, 다양한해조류를대상으로광범위한병원성세균및항생제내성균주에대한항균활성탐색평가는아직미미한상태이다. 본연구에서는 35 종의해조류추출물을이용하여병원성세균및항생제내성 Pseudomonas aeruginosa 및 Candida sp. 에대해 in vitro 항균활성을평가하였으며, 그결과패및넓패에서우수한항세균활성과항생제다제내성 Candida sp. 의제어가능성을확인하였기에이에보고하고자한다. 재료및방법 실험재료및시료의조제본실험에사용된해조류들은갈조류 17 종 ( 불레기말, 쇠미역, 감태, 곰피, 대황, 톳, 넓패, 패, 다시마, 알송이모자반, 모자반, 짝잎모자반, 괭생이모자반, 지충이, 야마다모자반, 고리매및미역 ), 홍조류 11 종 ( 돌가시리, 진두발, 도박, 참도박, 개도박, 얼룩도박, 방사무늬김, 작은구슬산호말, 붉은까막살, 마디잘록이및코도니 ) 및녹조류 7 종 ( 매생이, 청각, 잎파래, 납작파래, 구멍갈파래, 잘피및포도송이해초 ) 의 35 종으로 Table 1 에나타내었다. 이중코토니는인도네시아에서, 포도송이해초는베트남에서구입하여사용하였으며, 그외 33 종은국내부산, 경남, 전남등지에서구입하여사용하였다 [1]. 각각의해조류메탄올추출물은이미보고한바와동일하게제조하였다 [1]. 즉, 준비된해조류들은이물질을제거하고수돗물로세척한후실온에서음건하고이를믹서기로분쇄한후건조분말해조류를제조하였으며, 건조분말 100 g 당 1 리터의메탄올을가한후상온에서 2 일추출하고, 이를 4 회반복한후추출액을모아 50 o C 에서감압농축하여추출물을제조하였다. 조제된추출물은 dimethylsulfoxide(dmso) 를이용하여 100 mg/ml 농도로조정하여항균활성을평가하였다. 한편항균활성이우수한패, 넓패, 알쏭이모자반, 야마다모자반감태, 곰피, 대황및톳의 8 종의경우, 메탄올추출물로부터 n-hexane, ethylacetate, butanol 을이용하여순차적유기용매분획물과이후의물잔류물을조제한후, 각각의분획물을대상으로항생제내성 P. aeruginosa 및 Candida sp. 에대한항균활성을평가하였다. 사용균주본실험에사용한균주는그람음성세균 4종 (Pseudomonas aeruginosa, Escherichia coli, E. coli H7:O157, Salmonella typhimurium), 그람양성세균 2종 (Staphylococcus aureus, Table 1. The list of seaweeds used in this study. Scientific name Korean name Phaeophyta Colpomenia sinuosa Bulregi-mal Costaria costata Saemiyeuk Ecklonia cava Gamtae Ecklonia stolonifera Gompi Eisenia bicyclis (Kjellman) Setchell Daehwang Hixikia fusiforme Okamura Tot Ishige foliacea Nueppae Ishige okamurai Pae Laminaria japonica Areschong Dasima Sargassum confusum Alsongi-Mojaban Sargassum fulvellum Mojaban Sargassum hemiphyllum Jakip-Mojaban Sargassum horneri Gengsengi-Mojaban Sargassum thunbergii Jichungei Sargassum yamade Yamada-Mojaban Scytosiphon lomentaria Gorimae Undaria pinnatifida Miyeuk Rhodophyta Chondrus ocellatus Holmes Jindubal Corallina pilulifera Jaeungusel-Sanhomal Gigartina tenella Harvey Dolgasari Grateloupia elliptica Holmes Cham-Dovak Halymeniopsis dilatata Eulruk-Dovak Kappaphycus alvarezii Kotoni Lomentaria catenata Harvey Madi-Jalroki Pachymeniopsis elliptica Yamada Dovak Pachymeniopsis lanceolata Gae-Dovak Porphyra yezoensis Ueda Bansamuni-Kim Zanardinula cornea Bulkeun-Gamaksal Chlorophyta Capsosiphon fulvescens Maesengi Caulerpa racemosa Podosongi-Heacho Codium fragile (Suringar) Hariot Chungkak Enteromorpha compressa Napjak-Parae Enteromorpha linza Ip-Parae Ulva pertusa Gumunggal-Parae Zostera marina Jalpi Listeria monocytogenes) 및진균 3 종 (Candida albicans, Saccharomyces cerevisiae, Aspergillus niger) 을사용하였다 (Table 2). 그람음성세균중 P. aeruginosa 는항생제내성균주를포함한 7 종류, 진균중 Candida albicans 는항생제내성균주를포함한 7 종류를사용하였으며, 각각의미생물들은한국미생물자원센터 (KCTC), 한국농용미생물보존센터 (KACC), 일본 Institute for Fermentation, Osaka (IFO) 및한국항생제내성균주은행 (CCARM: Culture Collection of Antimicrobial Resistant Microbes) 에서분양받아사용하였다. P. aeruginosa CCARM 2010, 2020, 2030, 2200 및 2210 균주는 cefoperazone, gentamicin, norgloxacin, piperacillin,

146 KIM et al. Table 2. The microorganisms used in this study. Gram negative bacteria Gram positive bacteria Fungi Escherichia coli KCTC 1682 Escherichia coli H7:O157 ATCC 43895 Salmonella typhimurium KCTC 1926 Pseudomonas aeruginosa KACC 10186 Pseudomonas aeruginosa KACC 10259 Pseudomonas aeruginosa KACC 10260 Pseudomonas aeruginosa CCARM 2010 Pseudomonas aeruginosa CCARM 2200 Pseudomonas aeruginosa CCARM 2210 Pseudomonas aeruginosa PAO1 Staphylococcus aureus KCTC 1916 Listeria monocytogenes KACC 10550 Candida albicans KCTC 7270 Candida albicans KCTC 7965 Candida albicans KACC 30003 Candida albicans KACC 30004 Candida albicans KACC 30062 Candida albicans CCARM 14020 Candida albicans CCARM 14021 Candida kruseis CCARM 14017 Candida gillermordi CCARM 14018 Candida tropicalis CCARM 14019 Candida geochares KACC 30061 Candida saitoana KCTC 41238 Saccharomyces cerevisiae IF0 0233 Aspergillus niger KCCM 60143 amikacin, aztreoman, ceftazidime, cefotaxime 및 imipenem 의항생제중에서적어도 2종이상의항생제에대해내성을가지는다제내성균이며, 사람으로부터분리된균주이다. 한편 C. albicans CCARM 14020 및 14021, Candida kruseis CCARM 14017, Candida gillermordi CCARM 14018, Candida tropicalis CCARM 14019는 amphotericin B, itaconazole, flucytosine 및 fluconazole에대해다제내성을나타내는균주이다. 이들임상분리균주가상기의 β-lactam계 (ceftazidime, cefotaxime 등 ), aminoglycoside계 (gentamicin, amikacin 등 ), 세포막파괴제 (amphotericin B), nucleotide 유사체 (flucytosine) 등의항생제에내성을나타내는기작에대해서는현재까지알려진바없다. 항균활성평가항세균및항진균활성평가는기존의보고한 discdiffusion 방법 [26] 과동일하게이용하였으며, 세균의경우 Nutrient agar(difco Co., USA), 진균의경우 Sabouraud dextrose agar(difco Co. USA) 배지상에서다양한시료 5 µl 를멸균 disc-paper(whatsman No. 2, 지름 6.5 mm) 에가한후세균의경우 24 시간, 진균의경우 48 시간배양후, 생육억제환의크기를측정하여평가하였다 [26]. 특히 Asepergillus niger 에대한항균활성은 Potato dextrose broth(difco Co., USA) 를이용하여균사체생육억제활성을평가하였다. 대조구로는항세균제인 ampicillin 및항진균제 인 miconazole(sigma Co., USA) 를각각 1 µg/disc 농도로사용하였으며, 3 회이상평가후생육억제환지름의대표결과를 mm 단위로나타내었다 [27]. 인간적혈구용혈활성평가해조류추출물의실제적이용가능성평가의일환으로인간적혈구 (4% v/v) 를이용하여용혈활성을평가하였다 [27]. PBS 로 3 회수세한인간적혈구 100 µl 를 96-well microplate (Greiner Bio-one, flat bottom, Germany) 에가하고시료용액 100 µl 를가한다음 37 o C 에서 30 분간반응시켰다. 이후, 반응액을 10 분간, 1,500 rpm 으로원심분리 (HA-1000-3, Hanil Science Ind. Ltd. Korea) 하여상등액 100 µl 를새로운 microtiter plate 로옮긴후용혈에따른헤모글로빈유출정도를 microplate reader 기 (Sunrise-Basic, Tecan, Austria) 를이용하여 414 nm 에서측정하였다. 시료의용매대조구로는 DMSO(2%), 실험대조구로는 triton X-100 (0.1%) 을사용하였다. 용혈활성은다음의수식을이용하여계산하였다. (%) hemolysis = [(Abs. S Abs. C)/(Abs. T Abs. C)] 100. Abs. S: 시료첨가구의흡광도, Abs. C: DMSO 첨가구의흡광도, Abs. T: triton X-100 첨가구의흡광도 결과및고찰 35 종해조류의메탄올추출물을조제하여이들의항균활성을평가한결과는 Table 3 에나타내었다. 대조구로사용된항세균제 ampicillin 은 1 µg/disc 농도에서 P. aeruginosa, E. coli H7:O157 을제외한세균에대해우수한생육억제효과를나타내었으며, 항진균제 miconazole 은동일농도에서 S. cerevisiae, C. albicans 및 A. niger 에대해양호한항진균활성을나타내었다. 조제된해조류추출물들은 500 µg/disc 농도에서다양한병원성및식품부패균에대해각기다른항균스펙트럼을나타내었다. 먼저 P. aeruginosa 에대해항세균활성을나타내는해조류는없었다. E. coli 에대해서는넓패, 패, 다시마, 알쏭이모자반, 모자반, 짝잎모자반, 야마다모자반, 매생이및납작파래가우수한항균활성을나타내었고, 이중패에서생육억제환이 19.1 mm 로가장크게나타났다. 한편 E. coli H7:O157 에대해서는톳, 넓패, 패, 알쏭이모자반, 모자반, 짝잎모자반, 괭생이모자반, 지충이, 포도송이해초, 잎파래및구멍갈파래가항균성을나타내었고, 그중패와괭생이모자반에서 15.4 mm 이상의강한항균활성을보였다. 그러나 E. coli 에대한항균활성과일치하지는않았다. S. typhimurium 을대상으로항균력을검사한결과넓패, 패, 알쏭이모자반, 도박, 방사무늬김, 납작파래및잎파래에서항균활성을나타내었고, 잎파래의경우매우강력한생육억제력 (20.3 mm) 을나타내었다. S. aureus 에대해서는톳, 넓패,

Table 3. Antimicrobial and hemolytic activities of 35 different seaweeds extracts. Seaweeds Growth inhibition zone (mm) Gram negative Gram positive Fungi PA 1 EC EC 0157 ST SA LM CA SC AN BIOACTIVITY OF SEAWEEDS 147 Hemolytic activity against hrbc (%) 4 Phaeophyta Colpomenia sinuosa - 2 - - - - - - - - 2.0±1.3 Costaria costata - - - - - 8.0 - - - 3.0±2.9 Ecklonia cava - - - - - 9.0 7.0 - - 8.6±1.8 Ecklonia stolonifera - - - - - 9.0 7.0 - - 5.3±0.1 Eisenia bicyclis (Kjellman) Setchell - - - - - 9.0 7.0 - - 4.1±0.9 Hixikia fusiforme Okamura - - 11.7-12.6 9.5 7.5 - - 9.1±1.4 Ishige foliacea - 13.8 13.8 14.3 12.9 13.0 7.0 8.0-93.0±0.3 Ishige okamurai - 19.1 15.4 16.3 14.0 11.0 7.5 8.0-92.6±0.1 Laminaria japonica Areschong - 11.1 - - - 10.1 - - - 3.7±4.4 Sargassum confusum - 12.5 13.1 10.9 10.6 7.5 - - 0.2±0.7 Sargassum fulvellum - 10.1 15-11.7 - - - 5.7±5.4 Sargassum hemiphyllum - 16.6 14.8-16.3 - - - 1.2±1.8 Sargassum horneri - - 15.5-10.3 - - - 1.7±1.9 Sargassum thunbergii - - 12.9-12.7 10.1 - - - 0.5±0.8 Sargassum yamade - 15.6 - - 13.9 13.0 7.0 - - 55.9±5.1 Scytosiphon lomentaria - - - - - 9.5 - - - 9.5±0.2 Undaria pinnatifida - - - - - 7.5 - - - 4.7±1.8 Rhodophyta Chondrus ocellatus Holmes - - - - - - - - 1.5±1.8 Corallina pilulifera - - - - - 8.0 - - - 0.9±0.6 Gigartina tenella Harvey - - - - - - - - 1.8±0.9 Grateloupia elliptica Holmes - - - - - 8.0 - - - 0.7±0.4 Halymeniopsis dilatata - - - - - - - - - 0.6±0.1 Kappaphycus alvarezii - - - - - - - - - 0.9±0.9 Lomentaria catenata Harvey - - - - - - - - - 0.3±0.9 Pachymeniopsis elliptica Yamada - - - 9.0 - - - - - 0.3±0.2 Pachymeniopsis lanceolata - - - - - - - - - 0.3±0.5 Porphyra yezoensis Ueda - - - 9.0 - - - - - 0.9±0.4 Zanardinula cornea - - - - - - - - - 0.9±0.9 Chlorophyta Capsosiphon fulvescens - 9.8 - - - - - - - 0.1±0.2 Caulerpa racemosa - - 12.7 - - 12.5 - - - 20.3±0.1 Codium fragile (Suringar) Hariot - - - - - 9.0 - - - 1.1±0.2 Enteromorpha compressa - 9.2 13.3 10.1-10.0 - - - 28.8±3.8 Enteromorpha linza - - 17.4 20.3-10.0 - - - 3.7±7.2 Ulva pertusa - - - - - 9.0 - - - 3.7±4.3 Zostera marina - - - - - - -- - - 0.4±0.9 Positive control Ampicillin (1 µg/disc) - 11.0-10.0 21.0 19.0 - - - ND 3 Miconazole (1 µg/disc) - - - - - - 18.0 22.0 9.5 ND The concentrations of seaweed extracts used were 500 µg/disc, respectively. The hemolytic activity against hrbc was expressed as mean±standard deviation (n=3) 1 PA: Pseudomonas aeruginosa KACC 10186, EC: Escherichia coli, EC0157: Escherichia coli H7:O157, ST: Salmonella typhimurium, BS: Bacillus subtilis, SA: Staphylococcus aureus, LM: Listeria monocytogenes, CA: Candida albicans KCTC 7270, SC: Saccharomyces cerevisiae, and AN: Aspergillus niger. 2 -: No activity, 3 ND: Not determined, and 4 hrbc: human red blood cell.

148 KIM et al. 패, 알쏭이모자반, 모자반, 짝잎모자반, 괭생이모자반, 지충이, 야마다모자반추출물에서항균성을나타내었고, 그중짝잎모자반에서 16.3 mm 이상의강한생육억제활성을보였다. L. monocytogenes 에대해서는쇠미역, 감태, 곰피, 대황, 톳, 넓패, 패, 다시마, 지충이, 야마다모자반, 고리매, 미역, 작은구슬산호말, 참도박, 포도송이해초, 청각, 납작파래, 잎파래및구멍갈파래의 19 종의해조류가 7.5 mm 이상의생육억제환을나타내는항균활성을나타내었다. 전체적으로패와넓패가다양한세균에대해우수한항균활성을나타내었으며, 특히패의 S. aureus 에대한생육저해결과사진은 Fig. 1 에나타내었다. 한편캔디다증피부감염증의원인균인 C. albicans 에대해서는패, 넓패, 곰피, 대황, 감태, 톳, 알송이모자반및야마다모자반에서항진균활성이나타났으나, 그활성은생육억제환의크기가 7.0~7.5 mm 크기로미약하였다. 제빵및양조에사용되는 S. cerevisiae 에대해서는패와넓패에서만항진균활성이나타났으며, A. niger 에대해항곰팡이활성을나타내는해조류는없었다. 전반적으로홍조류보다는갈조류및녹조류추출물에서우수한항균활성이나타났으며, 특히넓패및패에서항진균활성이나타났다. 해조류추출물의실제적이용가능성과급성독성평가의일환으로각각의추출물을대상으로인간적혈구용혈활성을평가하였다. 500 µg/ml 농도에서 20% 이상의용혈활성을나타낸시료는넓패, 패, 야마다모자반, 포도송이해초및납작파래의 5 종이었다 (Table 3). 특히넓패, 패및야마다모자반은각각 93.01, 92.63 및 55.90% 의높은용혈활성을나 Fig. 1. Photography of cell growth inhibition in Ishige okamurai against Staphylococcus aurues. Symbols; M: methanol extract, H: hexane fraction, EA: ethylacetate fraction, B: butanol fraction, W: water residue of I. okamurai, and Amp: ampicillin as a positive control. Concentrations of samples of I. okamurai and ampicillin used were 250 µg/disc, and 1 µg/disc, respectively. The sizes of growth inhibition zone of M, H, EA, and ampicillin were 11.0, 14.3, 14.5, and 21.0 mm, respectively. 타내어이들추출물을식품에고농도로사용하는경우부작용이나타날가능성을예측할수있었다. 일반적으로항세균활성보다항진균활성을나타내는천연물의선별이보다필요하며, 특히항생제내성진균에대해항균활성을나타내는천연물은거의알려져있지않기때문에 C. albicans 에대해항진균활성을나타내는 8 종해조류는추가적으로순차적유기용매분획물을조제하여다양한항생제내성 C. albicans 및 Candida sp. 에대해항균력을평가하였다 (Table 4). 대조구로사용된 miconazole(1 µg/ disc) 은다양한항생제내성균주에대해 7.0~28.0 mm 의생육억제환을나타내었으나, C. albicans KCTC 7965, C. albicans CCARM 14020 및 CCARM 14021, C. krusei CCARM 14017, C. tropicalis CCARM 14019 에대해서는항균활성이나타나지않았다. 8 종해조류추출물및이들의각각의분획물들은 500 µg/disc 농도에서생육억제환을평가한결과, 강력한활성을나타내지는않았다. 먼저곰피의경우유기용매분획물에서는거의활성이나타나지않았고, 물잔류물에서만항진균활성이나타났다. 이는곰피의항진균활성성분이수용성물질임을의미하며, 차후곰피물잔류물에대한항진균활성물질탐색이필요하다고판단된다. 대황및감태의경우에는 butanol 분획물에서일부활성이나타났다. 한편패의경우, 다양한분획물에서항균활성이나타났으며, 특히 ethylacetate 분획및 butanol 분획은 C. albicans 6 종, Candida sp. 2 종및 S. cerevisiae 균주에대해항균력을나타내었다. 반면넓패의경우 hexane 분획물에서항균활성이나타났다. 톳의경우 butanol 분획및물잔류물에서주로활성이나타났으며, 알쏭이모자반의경우 hexene 분획을제외한다양한분획에서우수한활성이나타났다. 마지막으로야마다모자반의경우 butanol 분획및물잔류물에서항생제내성균주에대해항진균활성을나타내었다. 이러한결과는상기의 8 종해조류, 특히패및넓패로부터 C. albicans 및항생제내성 Candida sp 에대한신규의항균제개발가능성을제시하고있다. 한편상기 8 종해조류메탄올추출물및이들의각각의분획물을대상으로항생제내성균주를포함한다양한 P.aeruginosa 균주에대한항균력을평가하였으나, 시험균주모두에서항균활성이나타나지않았다 (result not shown). 본연구결과는해조류, 특히패, 넓패, 알송이모자반, 야마다모자반이다양한세균및항생제내성진균의제어에이용될수있는유용생물자원으로의가능성을제시하고있으며, 향후각각의해조류로부터실제적이용가능성검토를위한활성물질의규명및항균기작연구가필요하다. 요 해양생물유래의기능성물질개발연구의일환으로, 35 종해조류로부터메탄올추출물을조제하여항생제내성균 약

BIOACTIVITY OF SEAWEEDS 149 Table 4. Antifungal activity of the sequential organic solvent fractions and water residues of 8 different seaweeds against pathogenic Candida species and Sacchromyces cerevisiae. Extract and fractions of seaweeds Ecklonia stolonifera Eisenia bicyclis (Kjellman) Setchell Ecklonia cava Ishige okamurai Ishige foliacea Hixikia fusiforme Okamura Sargassum confusum Sargassum yamade Growth inhibition zone (mm) Candida albicans Candida sp. 1 1 2 3 4 5 6 7 8 9 10 11 12 M 3 7.0-4 - - - - - - - - - - - EA - - - - - - - - - - - - - B - - - - - - - - - - 8.0 - - W 7.0 - - 8.0 7.0-7.0 - - - 8.0 7.0 8.0 M 7.0 - - - - - - - - - 7.0 - - EA - - - - - - - - - - - - - B 7.0 7.0 7.0 7.0 - - 7.0 - - - - 7.0 7.0 W - - - - - - - - - - - - 7.0 M 7.0 - - - - - - - - - - - - EA - - - - - - - - - - - - B - - - - 7.0 - - - - 7.0 - - 7.0 W - - - - - - - - - - - - - M 7.5 - - - 7.5 - - - - - 8.0-8.0 H 8.0 - - - - 7.0 - - - - - - 7.0 EA 7.5-8.0 7.0 7.0-7.5 - - - 7.0 7.0 8.0 B 8.0 - - 8.0 8.0 7.0 8.0 - - - 8.0 9.0 - W - - - - - - 7.0 - - - - 7.0 - M 7.0-7.0 8.0 7.0 - - 8.0 - - 8.0-8.0 H - - 7.0 7.0 7.0 7.0 7.0 8.0 - - 7.0 7.0 - EA - - - - 7.0 - - - - - - - 7.0 B - - - - - - - - - - 7.5 - - W - - - - - - 7.5 - - - - 7.0 - M 7.5 - - - - - - - - - - - - EA - - 7.0 7.0 - - - - - - - - - B 7.5 - - 7.0 7.0 - - - - 7.0-7.5 7.0 W - - - - 7.0 - - - - 7.0 7.5 7.5 7.0 M 7.5 - - - - - - - - - - - - EA 7.5-8.0 - - 7.5 - - - - 7.5 8.0 7.5 B - - - 7.0 7.0 7.0 - - - 7.5-7.5 7.5 W - 7.0-7.0 7.0-7.5 - - - - - - M 7.0 - - - - - - - - - - - - H - - - - 7.5 - - - - - - 7.5 - EA - - 7.0 - - 7.0 - - - - - - - B 7.0 - - - 7.0 - - - - - - 8.5 7.0 W - - - - 7.0 7.0 - - - 7.0-7.5 7.0 Miconazole (1 µg/disc) 20.0-20.0 8.0 18.0 - - 28.0 7.0 11.0 - - 26.0 The concentrations of seaweed extracts and their fractions used were 500 µg/disc, respectively. 1 1: C. albicans KCTC 7270, 2: C. albicans KCTC 7965, 3: C. albicans KACC 30003, 4: C. albicans KACC 30004, 5: C. albicans KACC 30062, 6: C. albicans CCARM 14020, 7: C. albicans CCARM 14021. 8: C. geochares KACC 30061, 9; C. saitoana KACC 41238, 10: C. gillermordi CCARM 14018, 11: C. krusei CCARM 14017, and 12: C. tropicalis CCARM 14019. 2 SC: Saccharomyces cerevisiae. 3 M: Methanol extract, H: hexane fraction, EA: ethylacetate fraction, B: butanol fraction, and W: water residue, 4 -: No activity. SC 2

150 KIM et al. 주를포함한다양한세균및진균에대한항균력을평가하였다. 그결과, 패, 넓패, 알쏭이모자반및야마다모자반에서광범위한세균에대해우수한항세균활성을확인하였으며, 상기 4 종과함께곰피, 대황, 감태및톳의 8 종에서 Candida albicans 생육억제력을확인하였다. 이중패, 넓패, 야마다모자반추출물은 500 µg/ml 농도에서 55-93% 의인간적혈구용혈활성을나타내었다. 상기 8 종해조류의메탄올추출물을 n-hexane, ethylacetate, butanol 을이용하여순차적용매분획물과물잔류물을조제하여다양한 Candida sp. 에대한항균력을평가한결과, 패및넓패로부터 C. albicans 및항생제내성 Candida sp. 에대한신규의항균제개발가능성을확인하였다. 본연구결과는해조류특히패, 넓패, 알송이모자반, 야마다모자반이다양한세균및항생제내성진균의제어를위한생물자원으로개발가능함을제시하고있다. Acknowledgement This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0072444). REFERENCES 1. Ahn, S. M., Y. K. Hong, G. S. Kwon, and H. Y. Sohn. 2010. Evaluation of in vitro anticoagulation activity of 35 different seaweed extracts. J. Life Sci. 20: 1640-1647. 2. Bae, S. J. 2004. Studies on the antioxidative and antimicrobial effects of Chondria crassicaulis. J. Life Sci. 14: 411-416. 3. Bae, S. J. 2004. Anticarcinogenic effects of Sargassum fulvellum fractions on several human cancer cell lines in vitro. J. Korean. Soc. Food Sci. Nutr. 33: 480-486. 4. Cha, M. H. and Y. K. Kim. 2008. Analysis of consumption values of a seaweed functional food. Kor. J. Food Culture 23: 462-468. 5. Cho, E. K. and Y. J. Choi. 2010. Physiological activities of hot water extracts from Ecklonia cava Kjellman. J. Life Sci. 20: 1675-1682. 6. Choi, I. W., S. U. Kim, D. C. Seo, B. H. Kang, B. K. Sohn, Y. S. Rim, J. S. Heo, and J. S. Cho. 2005. Biosorption of heavy metals biomass of seaweeds, Laminaria species, Ecklonia stolonifera, Gelidium amansii, and Undaria pinnatifida. Kor. J. Environ. Agri. 24: 370-378. 7. Jin, H. J. and D. H. Jin. 2007. Screening of seaweed extracts for algicidal substances using a photosensitization effect. J. Kor. Fish Soc. 40: 122-127. 8. Joo, D. S., J. K. Lee, Y. S. Choi, S. Y. Cho, Y. K. Je, and J. W. Choi. 2003. Effect of sea tangle oligosaccharide drink on serum and hepatic lipids in rats fed a hyperlipidemic diet. J. Kor. Soc. Food Sci. Nutr. 32: 1364-1369. 9. Kang, M. C., J. Y. Lee, R. K. Ko, H. B. Kim, S. H. Hong, and G. O. Kim. 2008. Melanin inhibitory effect and antiinflammatory effects of Dietyota coriacea extracts derived from adjacent sea of the Jeju island. Kor. J. Biotechnol. Bioeng. 23: 311-316. 10. Kang, S. Y., M. J. Oh, and J. A. Shin. 2005. Antimicrobial activity of Korean marine algae against fish pathogenic bacteria. J. Fish Pathol. 18: 147-156. 11. Kim, B. J., S. H. Oh, and S. K. Park. 2006. Cross resistance between rifampicin and rofabutin and its relationship with rpob gene mutations in clinically isolated MDR-TB strains. Tuberc. Respir. Dis. 60: 171-179. 12. Kim, B. M., J. Y. Jun, Y. B. Park, and I. H. Jeong. 2006. Antioxidative activity of methanolic extracts from seaweeds. J. Kor. Soc. Food Sci. Nutr. 35: 1097-1101. 13. Kim, J. H., D. S. Lee, C. W. Lim, H. Y. Park, and J. H. Park. 2002. Antibacterial activity of sea-mustard, Laminaria japonica extracts on the cariogenic bacteria, Streptococcus mutans. J. Kor. Fish Soc. 35: 191-195. 14. Kim, S. A., J. Kim, M. K. Woo, C. S. Kwak, and M. S. Lee. 2005. Antimutagenic and cytotoxic effects of ethanol extracts from five kinds of seaweeds. J. Kor. Soc. Food Sci Nutr. 34: 451-459. 15. Kum, E. J., S. J. Park, B. H. Lee, J. S. Kim, K. H. Son, and H. Y. Sohn. 2006. Antifungal activity of phenanthrene derivatives from aerial bulbils of Dioscorea batatas Decne. J. Life Sci. 16: 647-652. 16. Lee, E. H., J. Ham, H. R. Ahn, M. C. Kim, C. Y. Kim, C. H. Pan, B. H. Um, and S. H. Jung. 2009. Inhibitory effects of the compounds isolated from Sargassum yezoense on - glucosidase and oxidative stress. Kor. J. Pharmacogn. 40: 150-154. 17. Lee, H. O., D. S. Kim, J. R. Do, and Y. S. Ko. 1999. Angiotensin-I converting enzyme inhibitory activity of algae. J. Kor. Fish Soc. 32: 427-431. 18. Lee, H. S., J. H. Suh, and K. H. Suh. 2000. Preparation of antibacterial agent from seaweed extract and its antibacterial effect. J. Kor. Fish. Soc. 33: 32-37. 19. Lim, C. W. 2000. Structure and some properties of the antimicrobial compounds in the red alga, Symphylocladia latiuscula. J. Kor. Fish Soc. 33: 280-287. 20. Lim, E. J., K. R. Cho, J. Y. Kim, Y. H. Lee, J. N. Ho, Y. J. Kim, and H. Y. Cho. 2008. The anticoagulant and anticancer activities of Enteromorpha interstinalis extracts. Kor. J. Food Nutr. 21: 7-14. 21. Lim, J. H., K. S. Jung, J. S. Lee, E. S. Jung, D. K. Kim, Y. S. Kim, Y. W. Kim, and D. H. Park. 2008. The study on antimicrobial and antifungal activity of the wild seaweeds of Jeju island. J. Soc. Cosmet. Sci. Kor. 34: 201-207. 22. Nikapitiya, C., M. Zoysa, P. M. Ekanyake, H. J. Park, and J. H. Lee. 2006. Isolation and purification of anticoagulant polysaccharide compound from fermented edible brown seaweed, Laminaria ochotensis. J. Aquaculture 19: 33-39.

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