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

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
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Structural Analysis of Anti-metastatic Polysaccharides Isolated from Opuntia humifusa

천년초에서 분리한 항전이 다당의 구조 분석

  • Choi, Jung-Ho (Dept. of Food Science & Biotechnology, Kyonggi University) ;
  • Shin, Kwang-Soon (Dept. of Food Science & Biotechnology, Kyonggi University)
  • 최정호 (경기대학교 식품생물공학과) ;
  • 신광순 (경기대학교 식품생물공학과)
  • Received : 2010.12.10
  • Accepted : 2010.12.15
  • Published : 2011.02.28
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Abstract

To examine the new practical utilization of mucilages in Opuntia humifusa, polysaccharides were isolated from O. humifusa and their anti-metastatic activity and structural analysis were carried out. In experimental lung metastasis of B16BL6 melanoma cells, prophylactically intravenous (i.v.) administration of the crude polysaccharide (CNC-0) from O. humifusa significantly inhibited lung metastasis in a dose-dependant manner. The main polysaccharide, CNC-Ia was purified to homogeneity from CNC-0 by two successive column chromatographies using DEAE-Sepharose FF and Sephadex G-100 and its structure was characterized. Molecular mass of CNC-Ia was estimated to be 700 kDa and it mainly consisted of arabinose, galactose and xylose in addition to two minor sugars such as rhamnose and fucose. Methylation analysis indicated that CNC-Ia comprised at least 18 different glycosyl linkages such as terminal Araf, 5-linked Araf, 4-linked Galp and terminal Xylp in addition to three characteristic linkages such as full branched Araf, 3,4,6-branched Galp and full branched Galp. To analyze the fine structure of CNC-Ia, it was sequentially digested by exo-${\alpha}$-L-arabinofuranosidase and endo-${\beta}$-1,4-D-galactanase. These analyses suggested that CNC-Ia belongs to be a highly branched Type I arabinogalactan which has a ($1{\rightarrow}4$)-${\beta}$-galactan backbone with arabinosyl oligosaccharide side chains.

천년초에 존재하는 점질다당의 새로운 이용방안을 모색하기 위해 천년초로부터 다당을 분리하여 항전이 활성을 평가하고 구조 분석을 행하였다. B16BL6 종양세포를 이용한 폐암 전이모델에서 천년초 조다당 CNC-0는 농도 의존적으로 높은 항전이 활성을 나타냈다. 천년초 조다당 CNC-0는 DEAE-Sepharose FF 및 Sephadex G-75를 이용한 연속적 chromatography를 행하여 CNC-Ia으로 정제하고 이들의 구조적 특성을 검토하였다. CNC-Ia는 분자량 약 700 kDa의 다당체로 구성당 조성을 확인한 결과, arabinose, galactose 및 xylose를 높은 비율로 함유하고 있었으며 rhamnose와 fucose를 미량 함유하고 있었다. 본 당쇄의 결합양식을 규명하기 위해 methylation analysis를 행한 결과 CNC-Ia은 terminal Araf, 5-linked Araf, 4-linked Galp, terminal Xylp와를 포함한 총 18종의 결합으로 구성되어 있었으며 full branched Araf, 3,4,6-branched Galp 및 full branched Galp 와 같은 3종의 CNC-Ia 고유의 특징적 결합을 포함하고 있었다. 또한 CNC-Ia의 미세구조를 규명하기 위해 exo-${\alpha}$-Larabinofuranosidase와 endo-${\beta}$-1,4-D-galactanase를 이용한 연속 가수분해 및 해석도 행하였다. 이상의 결과로부터 천년초 유래 다당 CNC-Ia는 ${\beta}$-($1{\rightarrow}4$)-galactan 주쇄에 arabinose oligo당이 측쇄로 분지된 Type I arabinogalactan으로 판단되었으며 주쇄 및 측쇄 모두 고도로 분지된 특징이 있음을 알 수 있었다.

Keywords

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