Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Isolation and Identification of an Antibacterial Substance from Sea Mustard, Undaria pinnatifida, for Streptococcus mutans

미역 추출물로부터 충치 원인균, Streptococcus mutans에 대한 항균물질의 분리 및 동정

  • Yun, So-Mi (Division of Marine Life Science and Technology, Institute of Marine Industry, Gyeongsang National University) ;
  • Jang, Jun-Ho (Graduate School of Agricultural Science, Tohoku Univ.) ;
  • Lee, Jong-Soo (Division of Marine Life Science and Technology, Institute of Marine Industry, Gyeongsang National University)
  • 윤소미 (경상대학교 해양생명과학부, 해양산업연구소) ;
  • 장준호 (일본동북대학 대학원 농학연구과) ;
  • 이종수 (경상대학교 해양생명과학부, 해양산업연구소)
  • Published : 2007.02.28
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Abstract

An antibacterial substance to the Streptococcus mutans, a causative bacterium for decayed teeth, was isolated from the dried sea mustard, Undaria pinnatifida, and identified by GC and GC/MS. Acetone extract from the sea mustard (10.4 kg), was evaporated and partitioned to 4 fractions such as hexane, chloroform, butanol and water. The most active chloroform fraction were further purified through basic alumina, silicic acid and ODS column, successively, and finally, 3 antibacterial substances were isolated on the HPLC attached ODS column by using 95% MeOH and guided with UV detector (254 nm). Antibacterial substances (total 160mg, yield $1.5\times10^{-3}$%) had the same Rf value (0.42) on the TLC developed hexane diethyl ether acetic acid (80:30:1) and those methyl esters moved to 0.95. They were identified as the same unsaturated fatty acid, $C_{18:4,\;n-3}$ (3,6,9,12-octadecatetraenoic acid, stearidonic acid) compared relative retention times (15.5 min) with authentic fatty acid on the GC chromatogram. It was further confirmed unambiguously on the GC/MS giving molecular ion peak at m/z 290 which coincided with its methyl ester.

건조 미역 10.4 kg을 acetone으로 추출, 여과한 다음 액-액 분배, 각종 크로마토그라피를 통하여 충치원인균, Streptococcus mutans에 대한 항균물질의 한 성분을 분리, 정제하고(160 mg, 수율 $1.5\times10^{-3}$%) 물질을 동정한 결과는 다음과 같다. 건조 미역 30 g 상당량의 추출액을 기준으로 용매 획분별 항균활성을 조사하였을 때, $CHCl_3$층이 73.2%로 가장 강하였고, hexane층이 62.0%이었으며, BuOH층이나 물층은 거의 없었다. 알루미나 칼럼에서는 산성 획분인 1% $NH_4OH:MeOH(1:1)$용매 획분이 81.0%의 항균활성을 나타내었으며, 실리카 칼럼에서는 $CHCl_3:MeOH(95:5)$용매에서 가장 높은 95.5%의 항균성을 나타내었고, ODS칼럼에서는 85% MeOH에서 96.4%의 항균성을 나타내었다. 최종적으로 ODS칼럼에서 95% MeOH를 이동상으로 하여 3개의 물질 S1(10 mg), S2(90 mg), S3(60 mg)을 분리 정제하였다. TLC에서 각 성분은 동일한 Rf값 0.42를 나타내어 동일한 물질로 추정되었으며, 이들을 메칠 유도체화한 성분들은 Rf값이 0.95로 바뀌어 이들 물질이 carboxyl기를 가지는 지방산으로 추정되었다. GC분석에서 표품 지방산과 비교한 결과, 이들은 $C_{18:4,n-3}$ 지방산과 retention time이 일치하였다. 또한, 메칠 유도체의 mass spectrum 분석 결과, m/z 290에 분자 이온 peak가 관측되어 $C_{18:4,n-3}$ 지방산의 methyl 유도체의 분자량과 일치하여, 이 물질을 3,6,9,12-octadecatetraenoic acid(stearidonic acid, $C_{18:4,n-3}$) 지방산으로 동정하였다.

Keywords

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