Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
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Purified Polysaccharide Activating the Complement System from Leaves of Diospyos kaki L.

감잎(Diospyos kaki L.)으로부터 정제한 보체계 활성화 다당류

  • Jung, Yung-Joo (Graduate School of Biotechnology, Korea University) ;
  • Chun, Hyug (Graduate School of Biotechnology, Korea University) ;
  • Kim, Kyung-Im (Graduate School of Biotechnology, Korea University) ;
  • An, Jeung-Hee (Graduate School of Biotechnology, Korea University) ;
  • Shin, Dong-Hoon (Graduate School of Biotechnology, Korea University) ;
  • Hong, Bum-Shik (Graduate School of Biotechnology, Korea University) ;
  • Cho, Hong-Yon (Graduate School of Biotechnology, Korea University) ;
  • Yang, Han-Chul (Graduate School of Biotechnology, Korea University)
  • 정영주 (고려대학교 생명공학원) ;
  • 전혁 (고려대학교 생명공학원) ;
  • 김경임 (고려대학교 생명공학원) ;
  • 안정희 (고려대학교 생명공학원) ;
  • 신동훈 (고려대학교 생명공학원) ;
  • 홍범식 (고려대학교 생명공학원) ;
  • 조홍연 (고려대학교 생명공학원) ;
  • 양한철 (고려대학교 생명공학원)
  • Published : 2002.10.01
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Abstract

Cold and hot water fractions of Diospyros kaki were screened to determine its anti-complementary activity. Flour of Diospyros kaki leaf (250 g) was boiled at $100^{\circ}C$ for 3 h and passed through a membrane of 10 kDa molecular weight (DK-0). DK-0 was precipitated with ethanol and refluxed with methanol to obtain the crude polysaccharide (DKC). DKC-1 was isolated by ion exchange chromatography on DEAE-Toyopearl 650C, and DKC-1c was purified from DKC-1 by size exclusion chromatography on Bio gel P-60. The anti-complementary activities of DKC-1c at $1000\;{\mu}g/mL$ were 85.4 and 61.1% via whole and alternative pathways, respectively. DKC-1c was determined as a neutral polysaccharide composed of glucose (29.0 mol.%), arabinose (24.3 mol.%), and galactose (16.2 mol.%) with the molecular weight of 66.6 kDa. Results of agarose gel immunoelectrophoresis revealed DKC-1c, as a complement activator, cleaved C3 into C3a and C3b via both pathways.

감잎으로부터 항보체 활성물질을 분리정제하기 위해 감잎 (250 g)을 $100^{\circ}C$에서 3시간 동안 열수 추출하고 분자량 10 kDa membrane을 사용하여 농축한 후 ethanol 침전과 methanol 추출을 통해 조다당류(DKC)를 얻었다. 조다당류의 정제는 DEAE-Toyopearl 650C와 Bio-gel P60을 사용하여 실시하였다. 정제된 DKC-1c는 $1000\;{\mu}g/mL$의 농도에서 고전경로를 통해서는 85.4% 활성화시켰고 부경로에서는 65.1% 활성화시켰다. 정제 다당류 DKC-1c는 분자량은 66.6 kDa이고 정제도가 높은 중성다당류로써 주요 구성당은 glucose(29.0 mol.%), arabinose(24.3 mol.%), galactose(16.2 mol.%) 순으로 검출되었다. 면역전기영동을 통하여 확인한 결과 DKC-1c는 C3를 부경로에서도 C3a와 C3b로 활성화시키는 complement activator임이 확인되었다.

Keywords

References

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