Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Isolation and Chemical Analysis of Potent Anti-Complementary Polysaccharides from Fruiting Bodies of the Fomes fomentarius

말굽버섯 자실체에서 분리한 항보체 활성 다당체의 화학적 분석

  • Park, Jung-Keun (Agricultural Biotechnology Department, National Academy of Agricultural Science, RDA) ;
  • Park, Kwe-Won (Department of Food Science & Biotechnology, Sunckyunkwan University) ;
  • Shin, Kwang-Soon (Department of Food Science & Biotechnology, Kyonggi University) ;
  • Lee, Chang-Muk (Agricultural Biotechnology Department, National Academy of Agricultural Science, RDA) ;
  • Seok, Soon-Ja (Agricultural Biology Department, National Academy of Agricultural Science, RDA) ;
  • Kim, Jeong-Bong (Agro-food Department, National Academy of Agricultural Science, RDA) ;
  • Koo, Bon-Sung (Agricultural Biotechnology Department, National Academy of Agricultural Science, RDA) ;
  • Han, Bum-Soo (Agricultural Biotechnology Department, National Academy of Agricultural Science, RDA) ;
  • Yoon, Sang-Hong (Agricultural Biotechnology Department, National Academy of Agricultural Science, RDA)
  • 박정근 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 박계원 (성균관대학교 식품생명공학과) ;
  • 신광순 (경기대학교 식품생물공학과) ;
  • 이창묵 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 석순자 (농촌진흥청 국립농업과학원 농업생물부) ;
  • 김정봉 (농촌진흥청 국립농업과학원 농식품자원부) ;
  • 구본성 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 한범수 (농촌진흥청 국립농업과학원 농업생명자원부) ;
  • 윤상홍 (농촌진흥청 국립농업과학원 농업생명자원부)
  • Received : 2012.11.01
  • Accepted : 2013.02.08
  • Published : 2013.06.28
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Abstract

The five anti-complementary polysaccharides (MFKF-NP, MFKF-AP1${\alpha}$, ${\beta}$, and MFKF-AP2${\alpha}$, ${\beta}$) were separated from hot water extracts of fruiting bodies of Fomes fomentarius by two subsequent column chromatography using DEAE-sepharose FF and Concanavalin A-sepharose 4B. The order of anti-complementary activity was MFKF-AP1${\beta}$ > MFKF-AP1${\alpha}$ > MFKF-AP2${\alpha}$ > MFKF-AP2${\beta}$ > MFKF-NP > Polysaccharide Krestine (PSK). Especially, MFKF-AP1${\beta}$ among those showed the most excellent anti-complementary activity (70% of ITCH50 value at $20{\mu}g/ml$). The monosaccharide composition analysis by gas chromatography indicates that MFKF-AP1${\alpha}$ and ${\beta}$ are a kind of homoxylan consisted mainly of xylose above 97%. Molecular weight of MFKF-AP1${\beta}$, major anti-complementary polysaccharide, was estimated to be about 12,000 by high performance liquid chromatography (HPLC). After the incubation of the serum with MFKF-AP1${\beta}$ in the presence or absence of $Mg^{++}$ and $Ca^{++}$ ions, its anti-complementary activity was investigated. This result indicated that MFKF-AP1${\beta}$ seems to be activator both on the classical and the alternative pathway of complement activation.

말굽버섯의 열수추출물에서 얻은 조다당체인 MFKF-CP를 분리, 정제하기 위하여 DEAE-sepharose FF 및 ConcanavalinA-sepharose 4B를 이용한 두 차례의 연속적인 chromatography를 수행하였다. DEAE-sepharose FF에서는 3종류의 다당체(MFKF-NP, MFKF-AP1, MFKF-AP2)를 분리하였는데, 그 중에서 MFKF-AP1이 $50{\mu}g/ml$ 농도에서 70% 이상의 높은 항보체 활성을 보였다. 이어서 ConcanavalinA-sepharose 4B 칼럼을 이용해 MFKF-AP1과 MFKF-AP2 다당체로부터 각각 MFKF-AP1${\alpha}$, ${\beta}$와 MFKF-AP2${\alpha}$, ${\beta}$ 다당체를 분리하였다. 그 중에서 MFKF-AP1${\beta}$가 $20{\mu}g/ml$ 농도에서 70% 이상의 가장 우수한 항보체 활성을 나타내었고 그 활성 순서는 MFKF-AP1${\beta}$ > MFKF-AP1${\alpha}$ > MFKF-AP2${\alpha}$ > MFKFAP2${\beta}$ > MFKF-NP > PKS이었다. 또한, $Mg^{++}$과 $Ca^{++}$ 이온이 제거되거나 첨가된 상태에서의 항보체 활성 실험을 통해 말굽버섯의 주 항보체 다당체인 MFKF-AP1${\beta}$는 고전경로(classical pathway)와 대체경로(alternative pathway) 모두를 경유하여 활성을 나타내고 있음을 확인하였다. Gas chromatography에 의한 구성당 조성 분석에서는 중성다당체인 MFKF-NP를 제외하고 나머지 4종의 다당체가 xylose를 약 70-99%의 높은 비율로 함유하고 있으며 특히 주요 항보체 다당체인 MFKF-AP1${\beta}$는 xylose를 99% 함유하고 glucose(0.24%) 및 arabinose (0.66%)를 미량 함유하고 있는 매우 특이한 homoxylan이었다. 또한, MFKF-AP1${\beta}$의 분자량을 HPLC로 분석한 결과, 약 12,000 정도인 것으로 추정되었다.

Keywords

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