Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Antiviral Activity of Hot-Water Extract and Its Ethanol Precipitate of Phellinus pini Fruiting Body

낙엽진흙버섯 자실체 유래 열수 추출물과 메탄을 침전물의 항바이러스활성

  • Lee, Sae-Mi (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Kim, Sung-Min (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Lee, Yoon-Hee (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Kim, Woo-Jung (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Na, Ye-Seul (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Kim, Hyun-Guell (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Nam, Jae-Hwan (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea) ;
  • Shin, Hyun-Dong (Dept. of Environmental Science and Ecological Engineering, Korea University) ;
  • Kwon, Doo-Han (Korea Research Institute for Bioscience and Biotechnology) ;
  • Park, Yong-Il (Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea)
  • 이새미 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 김성민 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 이윤희 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 김우중 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 나예슬 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 김현걸 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 남재환 (가톨릭대학교 생명공학과 & 생물소재공학연구센터) ;
  • 신현동 (고려대학교 환경생태공학과) ;
  • 권두한 (한국생명공학연구원) ;
  • 박용일 (가톨릭대학교 생명공학과 & 생물소재공학연구센터)
  • Published : 2009.03.28
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Abstract

The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an $EC_{50}$ (50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the $CC_{50}$ (50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.

알래스카산 낙엽진흙버섯[Porodaedalea pini (Brot.) Murrill] (syn. Phellinus pini)의 자실체 조분쇄물로부터 $100^{\circ}C$에서 4시간 추출하여 건량 수율 20.5%의 건조 분말 열수추출물을 제조하여, HeLa 세포에서 CVB3에 대한 항바이러스 활성을 조사한 결과 plaque 형성을 현저히 억제하였다. 또한, 다른 여러 가지 버섯 추출물에 비해 neuraminidase 활성 저해능이 가장 높았다. 열수 추출물로부터 75% 에탄올 침전으로 건량 수율 28.3%의 저분자 상등액 건조물(ES)과 수율 43.3%의 고분자 침전물 건조물(EP)을 얻었다. 상등액인 ES에서는 항바이러스 활성이 없었지만, 침전물인 EP는 HeLa 세포에서 CVB3바이러스에 대해 농도 의존적으로 plaque 형성을 현저히 억제하였고, $EC_{50}$은 0.45 mg/mL 이었으며, HeLa 세포에 대한 세포독성 $CC_{50}$은 2.25mg/mL 이었다. 또한, EP는 neureminidase 활성을 농도 의존적으로 저해하였으며, 1.7mg/mL에서 약 75%의 효소활성 억제효과를 보였다. 이러한 결과는 낙엽진흙버섯 자실체의 별수추출물로부터 얻어진 에탄을 침전물 EP가 CVB3 뿐만 아니라, influenza virus(Flu)등에 대해서도 광범위하게 항바이러스 활성을 나타낼 가능성을 보여 주었다. EP는 다당류로서 glucose의 함량이 79.8%로 가장 높았고 galactose, xylose, mannose와 fucose를 소량 포함한 수용성 heteroglycan의 일종으로, 소량(12.7%, w/w)의 단백질 또는 작은 펩타이드를 함유한 당단백의 일종일 것으로 사료된다.

Keywords

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