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1 The Korean Journal of Microbiology (2011) Vol. 47, No. 3, pp Copyright c 2011, The Microbiological Society of Korea 오염된음료에서분리된세균에대한한약재의항균성 유영은 1 김옥아 1 김상찬 1,2 박성민 1 * 1 대구한의대학교한방생명자원연구센터, 2 대구한의대학교한의학과 The Antibacterial Effects of Oriental Medicinal Herbs on Bacteria Isolated from Contaminated Beverages Young Eun Yu 2, Ok Ah Kim 2, Sang Chan Kim 1,2, and Sung Min Park 2 * 1 College of Oriental medicine, 2 The Research Center for Biomedical Resources of Oriental Medicine, Daegu Haany University, Daegu , Republic of Korea (Received August 25, 2011 / Accepted September 28, 2011) The use of synthetic additives for preservation and enhancement of the market quality of food products has been emerging as a societal issue in terms of safety as well as changes in consumption patterns. Various research related to natural additives is being conducted to address these issues. This study examined the antibacterial effects of 79 types of medicinal herbs used as oriental remedies on bacteria isolated from beverages of damaged marketable quality. The antibacterial effects of methanol extracts on 13 Bacillus sp. and three Paenibacillus sp. were evaluated. We found that 43 of the herbal medicines analyzed had antibacterial effects on the isolated bacteria. Of those, eight were selected, and their antibacterial effects were further examined using water, ethanol, methanol, and ethyl acetate as solvents. The results revealed that Prunus mume, Rhus javanica, and Coptis japonica had excellent antibacterial effects against the isolated bacteria. In particular, they exerted greater antibacterial effects when water and ethanol were used as solvents. This result indicates the possibility of developing natural additives using these substances. Since P. mume in particular, has not been sufficiently studied compared to other herbal medicines, it presents an opportunity for additional investigation and the possibility for development as a new product in the future. Keywords: antibacterial effect, natural additives, oriental medicinal herb 최근식품산업의추세는식생활의간편화와세대구성원의소수화로각종가공식품이나인스턴트식품의섭취가늘어나고있으며각종식품제조업체에서는다양한가공식품의제조에있어식품의가치를상승시키고소비자의구매욕구를충족시키기위하여다양한첨가제를사용하고있다 (5, 18). 식품제조업체에서더욱더다양한가공식품을제조함에따라제조과정중품질보존, 영양향상및관능적가치증진등을목적으로사용하는각종식품첨가물의소비량도증가하고있는추세이며, 이에각종가공식품에서다양한목적으로사용되고있는식품첨가물의섭취도불가피하게되어소비자가알지못하는사이에매일여러종류의식품첨가물을섭취하고있으며, 식품첨가물의안전성평가와섭취량에대한관심도증가하고있다. * For correspondence. minshell@dhu.ac.kr; Tel.: ; Fax: 이중식품보존제는가공식품의저장기간의연장을목적으로대부분의가공식품에사용할수밖에없으나사용되고있는합성식품보존제의경우인체의안전성에대한문제가사회적으로쟁점화되고있으며이에대한소비자들의소비습관에도많은영향을주고있다 (18). 이러한소비자들의소비습관에맞추어많은연구자들과기업에서제품의보존을위한소재를천연물로부터연구하고있으나관련시장에서는보다많은제품과다양한천연물에대한연구가절실한상황이다. 우리나라의경우전통의학인한의학과민간요법에서주로많이사용되는한약재와식물등의천연물을대상으로미생물에대한항균효능연구가많이진행되고있으며최근의연구를살펴보면단삼 (1), 매실 (3, 11), 인진쑥및녹차추출물 (4), 홍국 (5), 양파껍질 (6), 울금, 강황및보라울금 (7), 자몽종자추출물 (8, 21), 천연폴리페놀계물질을이용한연구 (9), 목초액 (12), 작약추출물 (16), 용아초추출물 (17), 산사자 (20), 다

2 242 Yu et al. 수의한약재 (2, 21, 24) 와천연물 (22), 프로폴리스 (23) 등을이용한연구가보고되어있으며 xylan과 chitosan을이용한보존제의개발 (13) 과오존수를이용한식품보존방법의개발 (15) 등많은연구가진행되고있다. 이에본연구자들은천연보존제로사용이가능한후보소재를발굴하기위하여오염된식품으로부터분리한세균을대상으로항균성을조사하였으며그결과를공유하고자한다. 재료및방법미생물의분리및선발오염된시료는경상북도영천시소재음료전문기업에서제품의제조후보관, 유통과정에서용기의형태변화나침전물 의발생등이야기되어회수한인삼추출물관련제품을사용하였다. 제품의제조전, 후에 80 C에서용기살균과정을거치는것을감안하여시료를 Purifier Biological Safety Cabinet (Labconco, USA) 에서멸균된용기에넣어끓는물에서 30 분간중탕하고실온에서식힌후 Nutrient medium (0.3% beef extract, 0.5% peptone, ph 6.8, Becton, Dickinson and Company, USA) 에단계희석도말하고 30, 37 C에서 48시간배양하여분리하였다. 염기서열분석및동정 SolGent TM Genomic DNA Prep Kit (SolGent, Korea) 을이용하여분리균주의 chromosomal DNA를분리하여 sequencing Table 1. List of oriental medicine herbs for this study Oriental Medicine Korean Name Use Part Oriental Medicine Korean Name Use Part Acanthopanax sessiliflorum Ogapi Root, Stem, Bark Lindera strichnifolia O-yag Root Agastache rugosa Gwaghyang Aerial part Lonicera japonica Geum-eunhwa Flower bud Ailanthus altissima Jeobaegpi Bark Lycium chinense Gugija Fruit Alpinia officinarum Golyanggang Rhizome Lycopus coreanus Taeglan Aerial part Amomum cadamomum Baegdugu Fruit Magnolia denudata Sin-ihwa Flower bud Amomum villosum Sa-in Fruit Melia japonica Golyeonpi Bark Aralia continentalis Doghwal Root Mentha piperascens Bagha Aerial part Areca catechu Daebogpi Pericarp Morus alba Sangbaegpi Bark Arisaema amurense Cheonnamseong Tuberous root Morus alba Sang-yeob Leaf Artemisia argyi Ae-yeob Leaf, Stem Nelumbo nucifera Yeonja-yug Seed Asiasarum mandshuricum Sesin Root, Rhizome Oldenlandia diffusa Baeghwasaseolcho All Brassica juncea Baeggaeja Seed Paeonia suffruticosa Mogdanpi Bark Bupleurum falcatum Siho Root Persicaria tinctoria Cheongdae Leaf Carpesium abrotanoides Hagseul Fruit Phellodendron amurense Hwangbaeg Bark Cassia tora Gyeolmyeongja Seed Polygala tenuifolia Wonji Root Chelidonium majus Baeggulchae Aerial part Polygonum aviculare Pyeonchug All Cibotium barometz Gucheog Rhizome Poncirus trifoliata Jisil Fruit Cinnamomum cassia Gyeji Branch Prunus mume Omae Fruit Citrus unshiu Jinpi Bark Pulsatilla koreana Baegdu-ong Root Clematis mandshurica Wilyeongseon Root Pyrrosia lingua Seog-wi Leaf Cnidium officinale Cheongung Rhizome Rehmannia purpurea Saengjihwang Root Codonopsis pilosula Mansam Root Rhus javanica Obaeja Cocoon Coptis japonica Hwanglyeon Rhizome Rosa rugosa Maegoehwa Flower bud Corydalis ternata Hyeonhosaeg Tuber Salvia miltiorrhiza Dansam Root Curcuma longa Ganghwang Root Saposhnikovia divaricata Bangpung Root, Rhizome Curcuma zedoaria Achul Rhizome Saururus chinensis Sambaegcho Aerial part Cynanchum atratum Baegmi Root Schizonepeta tenuifolia Hyeonggae Flower stalk Cyperus rotundus Hyangbuja Rhizome Scutellaria baicalensis Hwanggeum Root Dictamnus dasycarpus Baegseonpi Bark Smilax china Toboglyeong Root Drynaria fortunei Golswaebo Rhizome Sophora japonica Goehwa Flower bud Eclipta prostrata Hanlyeoncho All Spatholobus suberectus Gyehyeoldeung Stem Equisetum hyemale Mogjeog Aerial part Stemona japonica Baegbugeun Tuberous root Foeniculum vulgare Hoehyang Fruit Tamarix juniperina Jeonglyu Branch, Leaf Ganoderma lucidum Yeongji Fruit body Thuja orientalis Baegja-in Seed Gentiana macrophylla Jingyo Root Tribulus terrestris Jillyeoja Fruit Gingko biloba Baeggwa Seed Uncaria sinensis Jogudeung Branch Glycine max Daeduhwanggwon Germination Vitex rotundifolia Manhyeongja Fruit Isatis indigotica Daecheong-yeob Leaf Xanthium strumarium Chang-ija Fruit Juncus effusus Deungsimcho Stem Zingiber officinale Saenggang Rhizome Leonurus sibirieus Chung-wija Fruit

3 식품보존제의개발을위한한약재의항균성조사 243 에사용하였다. Initial denaturation 94 C, 3분, denaturation 94 C, 1분, annealing 55 C, 1분, extension 72 C, 3분의조건으로 32 cycle을실시한후, final extension을 72 C에서 5분간유지하여증폭을종결하였다. 16S rrna의증폭을위하여 E. coli 16S rrna 부분의 conserved sequence를기초로하여합성된 universal primer인 27F (5 -AGAGTTTGATCATGGGT CAG-3 ) 와 1492R (5 -GGATACCTTGTTACGACTT-3 ) 를 primer로사용하였다. 16S rrna의 PCR 산물은 SolGent TM PCR Purification Kit (SolGent, Korea) 로정제하였고정제된 PCR 산물은 ABI PRISM 3700 DNA Analyzer를이용하여염기서열을분석한후, BLASTN 프로그램을이용하여 GenBank의 ribosomal RNA sequence와비교하여동정하였으며대전광역시에위치한 ( 주 ) 솔젠트에의뢰하여분석하였다 (19). G+C 함량등보다구체적인동정과정을진행하지않았기에 16S rrna의분석결과는대략적인균주를파악하기위해서만사용하였다. 한약재의선발오염된식품으로부터분리한세균에대한항균성을나타내는한약재를조사하기위하여본초학을바탕으로문헌조사를하였으며이를바탕으로한약재를선발하여수성구상동소재의대원약업사에서생약제제로포장되어판매하는제품을구입하여사용하였다 (Table 1). 추출물의제조분리세균에대한항균성을조사하기위하여한약재 79종을선발하였으며일차조사는메탄올을추출용매로사용하고, 이차조사는물, 에탄올, 메탄올및에틸아세테이트를사용하여추출물을제조하여사용하였다. 추출물의제조에있어에탄올, 메탄올및에틸아세테이트를사용한경우한약재에 10배량의추출용매를가하고상온에서 150 rpm, 24시간동안진탕하고여과 (Filter paper No. 2, Whatman, Japan) 한후대형회전농축기 (Rotary evaporator, Switzerland) 로감압농축하고각각의추출용매로다시현탁한후 0.2 μm filter (Minisart RC 25, Sartorius, Germany) 로제균하여사용하였다. 열수추출물의경우에는 10배량의 1차정제수를가하여약탕기로 96 C에서 2시간정도끓여추출하였으며다른추출물과동일한방법으로준비하여사용하였다. 추출물의항균성조사항균성을조사하기위하여분리한세균을 24시간배양한후원심분리 (13,000 rpm, 2 min) 하고멸균수로현탁시키는과정을 3번반복하여준비하였으며 1.5 ± cells/ml 로균수를조정하여사용하였다. 선발한 79종의한약재중항균성을나타내는한약재를선별하기위하여먼저메탄올추출물을이용하였으며그결과를바탕으로한약재를선별한후용매별항균성을조사하였다. 추출물의농도는회수율을적용하였으며한약재의추출에 사용한용매로각각의추출물을녹여사용하였다. 즉오매메 탄올추출물의경우 22.1% 의회수율을나타내어 g의 추출물을 1 ml의메탄올에녹여샘플을제조하였다. 두번의조사는추출물을 6 mm paper disc (Advantec, Japan) 를사용한한천평판확산법 (Agar diffusion method) 을이용하 였으며한약재의추출에사용한용매로각각의추출물을녹여 사용하였다. 분리세균의생육이억제되면서형성되는생육저지환 (Clear zone) 의크기로항균성의유ㆍ무와정도를확인하였으며 3번 반복실험을한후평균값과표준편차를구하였으며편차의정 도가미비하여평균값만을표기하였다. 결과및고찰 분리세균의동정및선발한약재의회수율 오염된음료로부터분리된세균은 Table 2와같이 13종의 Bacillus sp. 와 3종의 Paenibacillus sp. 로판명되었고상동성 도높게조사되었으나앞서언급한것처럼명확한동정을진 행하지않아속명에번호를붙여사용하였다. 분리세균들은 고체배지와액체배지의배양을통하여 37 C에서양호하게생 육하는것을확인하였으며배양조건을 37 C로통일하여적 용하였다 ( 결과미제시 ). 추출용매로메탄올을사용한경우의회수율은백부근이 59.6% 로가장높게나타났으며백두옹외 6종이 20% 이상 30% 미만을, 금은화외 30종이 10% 이상 20% 미만을, 박하 외 37종이 1% 이상 10% 미만을나타내었으며청대와결명자 의경우 0.1% 의매우낮은회수율을나타내는것으로조사되 었으며선발한한약재의 50% 정도가 10% 미만의낮은회수 율을나타내었다. 나등 (14) 이결명자를물로추출하였을때 20% 이상의회수율을나타낸결과와비교할때본연구자들 Table 2. Isolated bacteria from contaminated drink No. Genus Species Max identity (%) 1 Bacillus amyloliquefaciens 98 2 Bacillus subtilis 98 3 Paenibacillus chibensis 98 4 Bacillus circulans 95 5 Paenibacillus soli 99 6 Bacillus circulans 98 7 Bacillus amyloliquefaciens 99 8 Bacillus circulans 99 9 Bacillus ginsengihumi Bacillus amyloliquefaciens Paenibacillus chibensis Bacillus amyloliquefaciens Bacillus mojavensis Bacillus ginsengihumi Bacillus circulans Bacillus benzoevorans 97

4 244 Yu et al. 이확인한회수율 0.1% 와는많은차이를나타내었으며이는결명자의경우추출용매에대한영향을많이받는것을확인할수있었다. 추출물의회수율은회수된추출물의중량을추출시사용한한약재의중량으로나눈후백분율로계산하였으며소수점첫째자리까지기록하였다. 한약재메탄올추출물의항균성항균성을조사하기위하여미리배양해둔분리세균의수를 1.5 ± cells/ml로조정한후 100 μl를첨가하여고체배지를제조한후각각의메탄올추출물 20 μl를 paper disc에점적하여상온에서건조시키고배지에얹어 37 C에서 24시간배양하여조사한항균효과의결과는 Table 3과같이조사되었다. 선발한 79종의한약재중진피, 황련, 등심초, 목단피, 청대, 오매, 오배자, 매괴화, 단삼과괴화의메탄올추출물은모든공시균주에대하여항균성을나타내는것으로조사되었으며고량강, 학슬, 상백피, 황백, 황금과질려자의경우에도 15종에대하여항균성을나타내는것으로조사되었다. 그러나대복피, 백개자, 결명자, 골쇄보, 연자육과생지황의경우공시균주에대한항균성을전혀나타내지않는것으로조사되었다. 각각의공시균주에대한메탄올추출물의항균성은 Bacillus sp. CBB1과 Bacillus sp. CBB2의경우오매추출물이각각 19와 20 mm로가장우수하였으며 Paenibacillus sp. CBB3의경우에도오매추출물이 25 mm로가장우수하였고세신, 황련과오배자추출물도 20 mm 이상의생육저지환을형성하였다. Bacillus sp. CBB4의경우애엽과오매추출물이가장우수하였고 (23 mm) 황련, 오배자와질려자추출물도 20 mm 이상의생육저지환을형성하였다. Paenibacillus sp. CBB5의경우오배자추출물이 25 mm 로가장우수하였으며세신, 학슬, 계지, 회향, 등심초, 택란과오매추출물도 20 mm 이상의생육저지환을형성하였다. Bacillus sp. CBB6의경우오매추출물이 22 mm로가장우수하였으며오배자추출물도 21 mm의생육저지환을형성하였다. Bacillus sp. CBB7의경우황련추출물이 22mm로가장우수하였으며황백추출물은 21 mm, 지실추출물은 20 mm 의생육저지환을형성하였고 Bacillus sp. CBB8의경우오매추출물만 20 mm 이상의생육저지환을형성하였다. Bacillus sp. CBB9의경우애엽, 황련과회향추출물이 22 mm로가장우수하였으며황백, 지실과질려자추출물도 20 mm 이상의생육저지환을형성하였다. Bacillus sp. CBB10의경우오매추출물이 15 mm로가장우수하였으나공시균주중추출물에의한생육의저해가미비한것으로나타났으며 Paenibacillus sp. CBB11의경우오매추출물이 23 mm로가장우수하였으며황련과계혈등추출물에서도 20 mm 이상의생육저지환을형성하였다. Bacillus sp. CBB12와 CBB13의경우오매와오배자추출 물이 15 mm로가장우수한생육저지환을생성하였으나 Bacillus sp. CBB10의경우처럼항균성은다른한약재추출물과비교할때낮은것으로조사되었다. Bacillus sp. CBB14의경우황련추출물이 26 mm로가장우수하였으며오가피, 애엽, 학슬, 향부자, 회향, 택란, 상엽, 황백, 석위, 오배자와괴화추출물에서도 20 mm 이상의생육저지환을형성하여조사한결과중가장많은한약재에의하여생육의저해를받는것으로나타났다. Bacillus sp. CBB15의경우황련추출물이 23 mm로가장우수한것으로조사되었으며애엽, 목적, 지실과석위추출물에서도 20 mm 이상의생육저지환을형성하였으며 Bacillus sp. CBB16의경우황련추출물이 22 mm로가장우수하였고오배자추출물도 20 mm 이상의생육저지환을형성하였다. 선발한한약재중 43종이 50% 이상의공시균주에대한항균성을나타내는것으로조사되어한약재의선발을위한문헌적조사결과가어느정도양호하였음을확인할수있었다. 용매에따른추출물의항균성메탄올추출물에의한항균성을바탕으로공시균주에대하여넓은항균스펙트럼과상대적으로우수한항균성을가지는한약재 8종을선별하고물, 에탄올, 메탄올과에틸아세테이트를추출용매로하여각각의추출물을제조한후항균성을조사하였다 (Table 4.). 한약재의회수율의경우우선물을추출용매로사용한열수추출의경우오배자가 71.2% 로가장높은회수율을나타내었으며청대가 1.3% 로가장낮은회수율을나타내었다. 에탄올을사용한경우에는오매가 24.8% 로가장높은회수율을나타내었으며청대가 0.7% 로가장낮았고, 메탄올을사용한경우오매가 28.9% 로가장높은회수율을나타내었으며청대가 1.0% 로가장낮았으며에틸아세테이트의경우오배자가 13.0% 로가장높은회수율을나타내었으며황련이 0.2% 로가장낮은회수율을나타내었다. 열수추출물의경우오매, 오배자와황련의경우에만공시균주에대하여전반적인항균성을나타내었으며오매가가장양호한항균성을나타내는것으로조사되었다. 나머지 5종의한약재의경우항균성이미비하거나항균성을나타내지못하였으며특히등심초와지실의경우모든공시균주에대하여항균성을나타내지않았다. 에탄올추출물의경우괴화, 단삼, 오매와오배자의경우에모든공시균주에대한항균성을나타내었으며오매가가장양호한항균성을나타내는것으로판단되었다. 메탄올추출물의경우 8종의한약재가대부분의공시균주에대하여항균성을가지는것으로조사되었으며앞선조사의회수율과생육저지환의크기에서약간의차이는발생하였으나이는추출물을준비하는과정에서발생할수있는것이라판단되었다. 에틸아세테이트추출물의경우괴화에서대부분의공시균주에대한항균성이조사되었으나다른추출물과비교할때항균성은상대적으로미비한것으로판단하였다. 그러나낮은

5 식품보존제의개발을위한한약재의항균성조사 245 Table 3. Antibacterial activity of oriental medicine herbs extracted with methanol No. Name Recovery rate (%) A. sessiliflorum A. rugosa A. altissima A. officinarum A. cadamomum A. villosum A. continentalis A. catechu A. amurense A. argyi A. mandshuricum B. juncea B. falcatum C. abrotanoides C. tora C. majus C. barometz C. cassia C. unshiu C. mandshurica C. officinale C. pilosula C. japonica C. ternata C. longa C. zedoaria C. atratum C. rotundus D. dasycarpus D. fortunei E. prostrata E. hyemale F. vulgare G. lucidum G. macrophylla G. biloba G. max I. indigotica J. effusus L. sibirieus L. strichnifolia L. japonica L. chinense L. coreanus M. denudata M. japonica M. piperascens M. alba M. alba N. nucifera O. diffusa P. suffruticosa P. tinctoria P. amurense

6 246 Yu et al. Table 3. Continued No. Name Recovery rate (%) P. tenuifolia P. aviculare P. trifoliata P. mume P. koreana P. lingua R. purpurea R. javanica R. rugosa S. miltiorrhiza S. divaricata S. chinensis S. tenuifolia S. baicalensis S. china S. japonica S. suberectus S. japonica T. juniperina T. orientalis T. terrestris U. sinensis V. rotundifolia X. strumarium Z. officinale : < 10 mm, ++ : < 20 mm, +++ : < 30 mm 1. Bacillus sp. CBB1, 2. Bacillus sp. CBB2, 3. Paenibacillus sp. CBB3, 4. Bacillus sp. CBB4, 5. Paenibacillus sp. CBB5, 6. Bacillus sp. CBB6, 7. Bacillus sp. CBB7, 8. Bacillus sp. CBB8, 9. Bacillus sp. CBB9, 10. Bacillus sp. CBB10, 11. Paenibacillus sp. CBB11, 12. Bacillus sp. CBB12, 13. Bacillus sp. CBB13, 14. Bacillus sp. CBB14, 15. Bacillus sp. CBB15, 16. Bacillus sp. CBB16. 회수율에도불구하고다른추출물과유사한정도의항균성을나타내는경우도있음으로볼때추출물의회수율과기대한항균성과의상관관계는미비한것으로판단되었다. 황련의경우이등 (10) 도식품부패균에대한항균성을보고하였으며오배자의경우에도어병세균에대하여가장우수한항균성을나타내었다고보고되어있어다른연구자들의연구결과와본연구의결과를바탕으로오매, 오배자와황련은음료의보존을위한소재로사용할수있을것으로판단하였다. 특히오매의경우다른한약재와비교할때미생물에대한항균성연구가미진한것으로판단되었으며천연식품보존제개발을위한소재로서기존에알려진오배자와황련보다다양한적용이가능할것으로판단하였다. 향후본연구자들은다양한식품오염미생물을대상으로연구를진행하여한약재를기반으로한천연식품보존제의개발가능성을조사하고자한다. 적요식품의상품성과보존을위하여사용하는합성첨가물이소비자들의소비습관의변화와첨가물의안전성에대한사회적쟁점화로천연의소재를활용한다양한연구와조사가진 행되고있다. 본연구에서도천연물중한의학의소재로사용되는생약제제 79종을사용하여식품중상품성이훼손된음료에서분리한세균에대한항균성을조사하였다. 13종의 Bacillus sp. 와 3종의 Paenibacillus sp. 에대한메탄올추출물의항균성을조사한결과 43종의생약제제가분리한세균에대한항균성을나타내었다. 이중 8종의한약재를선별하고이들을대상으로물, 에탄올, 메탄올과에틸아세테이트를추출용매로하여항균성을조사한결과오매, 오배자와황련이분리된세균에대한항균성이우수한것으로조사되었으며특히물과에탄올을사용한경우에우수한항균성을나타내어이결과를바탕으로이들소재를활용한천연보존료의개발가능성을확인할수있었다. 특히다른생약제제와비교할때오매에대한연구가상대적으로미진한것으로판단되어향후지속적인연구를통한제품의개발도가능할것으로판단하였다. 감사의말본연구는지식경제부의지역혁신센터사업을수행하고있는대구한의대학교한방생명자원연구센터 (B ) 의지원에의하여이루어졌으며이에감사드립니다.

7 식품보존제의개발을위한한약재의항균성조사 247 Table 4. Antibacterial activity of selected oriental medicine herbs extracted with 4 solvents No. Name Solvents Recovery rate (%) Water C. japonica EtOH MeOH Ethylacetate Water J. effusus EtOH MeOH Ethylacetate Water P. tinctoria EtOH MeOH Ethylacetate Water P. trifoliata EtOH MeOH Ethylacetate Water P. mume EtOH MeOH Ethylacetate Water R. javanica EtOH MeOH Ethylacetate S. miltiorrhiza 8 S. japonica Water EtOH MeOH Ethylacetate Water EtOH MeOH Ethylacetate 참고문헌 1. Choi, H.Y Antimicrobial activity of Salvia miltiorrhiza Bunge extract and Its effects on quality characteristics in sulgidduk. Korean J. Food & Nutr. 22, Choi, H.S., Y.C. Kim, J.S. Lee, M.R. Jo, C.H. Seo, and S.I. Park Antibacterial activity of hot-water and ethyl alcohol extracts of medicinal herbs on fish pathogenic bacteria. J. Fish Pathol. 17, Ha, M.H. and S.H. Cho Preservative effect of agricultural and marine products treated with Prunus mune extract. J. Agriculture & Life Sciences 39, Kim, D.W., J.H. Kim, G.H. Kang, H.K. Kang, S.B. Park, J.H. Park, H.T. Bang, M.J. Kim, J.C. Na, H.S. Chae, and et al Studies for antibiotics free chicken production using water extracts from Artemisia capillaries and Camellia sinensis. Korean J. Food Sci. Ani. Resour. 30, Kim, E.Y. and M.R. Rhyu Antimicrobial activities of Monascus koji extracts. Korean J. Food Sci. Technol. 40, Kim, J.E., A.R. Kim, M.J. Kim, and S.N. Park Antibacterial, antioxidative and antiaging effects of Allium cepa peel extracts. Appl. Chem. Eng. 22, Kim, H.J., J.W. Lee, and Y.D. Kim Antimicrobial activity and antioxidant effect of Curcuma longa, Curcuma aromatic and Curcuma zedoaria. Korean J. Food Preserv. 18, Kim, H.S., J.W. Park, I.B. Park, Y.J. Lee, J.M. Kim, and Y.C. Jo Optimization of antimicrobial activity against foodborne pathogens in grapefruit seed extract and a lactic acid mixture. Korean J. Food Preserv. 16, Kim, T.K., H.E. Shin, and Y.H. Lee Stabilization of polyphenolic antioxidants using inclusion complexation with cyclodextrin and their utilization as the fresh-food preservative. Korean J. Food Sci. Technol. 35, Lee, G.H., E.S. Doh, J.P. Chang, and G.J. Kil Antibacterial activity of Caesalpinia sappan and Coptis chinensis extracts against Bacillus cereusand Vibrio parahaemolyicus. Kor. J. Herbology 25, Lee, H.A., E.S. Nam, and H.I. Park Antimicrobial activity of Maesil (Prunus mume) juice against selected pathogenic microorganisms. Korean J. Food & Nutr. 16, Lee, S.S., B.J. Ahn, and S.T. Cho Antimicrobial activities of wood vinegar and application as natural fungicides and food preservatives. Mokchae Konghak 38, Li, X., X. Shi, M. Wang, and Y. Du Xylan chitosan conjugate A potential food preservative. Food Chemistry 126,

8 248 Yu et al. 14. Na, G.M., H.S. Han, S.H. Ye, and H.K. Kim Extraction characteristics and antioxidative activity of Cassia tora L. extracts. Korean J. Food Culture 19, Park, J.M. and H.S. Shin Influence of ozonated water and washing method using ozonated water for controlling food-borne disease bacteria. Food Engineering Progress 14, Park, K.D. and S.H. Cho Antimicrobial characteristics of Paeonia lactiflora Pall. extract tested against food-putrefactive microorganisms. Korean J. Food Preserv. 17, Park, N.Y., K.N. Park, and S.H. Lee Antimicrobial activities and food preservative effects of Agrimoniae herba. J. Korean Soc. Food Sci. Nutr. 33, Park, S.K., E.S. Lee, and S.K. Park Estimation of daily dietary intake of food red colors food red No. 2, No. 3 and No. 40 J. Korean Soc. Food Sci. Nutr. 34, Park, S.M., J.S. Lee, C.D. Park, J.H. Lee, H.J. Jung, and T.S. Yu Selection and antifungal activity of antagonistic bacterium Bacillus subtilis KMU-13 against cucumber scab, Cladosporium cucumerinum KACC Korean J. Biotechnol. Bioeng. 21, Ryu, H.Y., S.M. Ahn, J.S. Kim, I.C. Jung, and H.Y. Sohn Antimicrobial activity of fruit of Crataegus pinnatifida Bunge against multidrug resistant pathogenic Pseudomonas aeruginosa and Candida sp. Kor. J. Microbiol. Biotechnol. 38, Ryu, H.Y., S.M. Ahn, Y.K. Shin, and H.Y. Sohn Antimicrobial and hemolytic activity of oriental medicinal herbs. Kor. J. Microbiol. Biotechnol. 38, Tak, S.B., D.H. Kim, S.K. Yoon, and Y.C. Lee Effects of natural preservatives and storage temperatures on quality and shelf-life of fresh pork meat. Korean J. Food Sci. Technol. 37, Tosi, E.A., E. Re, M.E. Ortega, and A.F. Cazzoli Food preservative based on propolis: Bacteriostatic activity of propolis polyphenols and flavonoids upon Escherichia coli. Food Chemistry 104, Yu, Y.E., E.Y. Park, D.H. Jung, S.H. Byun, S.C. Kim, and S.M. Park Antibacterial effect of oriental herbs on dental pathogens. Kor. J. Microbiol. 46,

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14_HM hwp The Korean Journal of Microbiology (2010) Vol. 46, No. 2, pp. 200-206 Copyright c 2010, The Microbiological Society of Korea 구강질환세균에대한한약재의항균효과 유영은 1 박은영 1 정대화 1 변성희 2 김상찬 1,2 박성민 1 * 1 대구한의대학교한방생명자원연구센터,