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 of Polysaccharides Modulating Mouse’s Intestinal Immune System from Peels of Citrus unshiu

귤피로부터 분리한 마우스의 장관면역 활성 다당류의 검색

  • Yang, Hyun-Seuk (Food Safety Research Center, CJ Corporation) ;
  • Yu, Kwang-Won (Dept. Kimchi and Food Science, Chongju National College of Science and Technology) ;
  • Choi, Yang-Mun (Dept. Food Service Industry, Shinsung College)
  • 양현석 (CJ(주) 식품안전연구센터) ;
  • 유광원 (청주과학대학 김치식품과학과) ;
  • 최양문 (신성대학 호텔식품계열)
  • Published : 2004.11.01
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Abstract

Of solvent-extracts prepared from the 90 kinds of Korean traditional tea and rice gruel plants, cold-water extract from peels of Citrus unshiu (CUI-0) showed the most potent intestinal immune system modulating activity through Peyer’s patch whereas other extracts did not have the activity except for cold-water extracts of Laminaria japonica, Polygonatum japonicum, Poncirus trifoliata, and hot-water extracts of Gardenia jasminoides, Lycium chinense having intermediate activity. CUI-0 was further fractionated into MeOH-soluble fraction (CUI-1), MeOH insoluble and EtOH-soluble fraction (CUI-2), and crude polysaccharide fraction (CUI-3). Among these fractions, CUI-3 showed the most potent stimulating activity for the proliferation of bone marrow cells mediated by Peyer’s patch cells, and contained arabinose, galacturonic acid, galactose, glucose, glucuronic acid and rhamnose (molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) as the major sugars, and a small quantity of protein (9.4%). In treatments of CUI-3 with pronase and periodate (NaIO₄), the intestinal immune system modulating activity of CUI-3 was significantly reduced, and the activity of CUI-3 was affected by periodate oxidation particularly. The potently active carbohydrate-rich fraction, CUI-3IIb-3-2 was further purified by anion-exchange chromatography on DEAE-Sepharose FF, Sepharose CL-6B and Sephacryl S-200. CUI-3IIb-3-2 was eluted as a single peak on HPLC and its molecular weight was estimated to be 18,000 Da. CUI-3IIb-3-2 was consisted mainly of arabinose, galactose, rhamnose, galacturonic acid and glucuronic acid (molar ratio;1.00:0.54:0.28:1.45:0.63) in addition to a small amount of proteins (3.2%). In addition, CUI-3IIb-3-2 showed the activity only through Peyer’s patch cells, but this fraction did not directly stimulate proliferation of bone marrow cells. It may be concluded that intestinal immune system modulating activity of peels from C. unshiu is caused by pectic polysaccharides having a polygalacturonan moiety with neutral sugars such as arabinose and galactose.

90여종의 전통차 및 죽류용 식물로부터 조제된 다양한 용매추출물 중에서, 귤피(peels of C. unshiu)의 냉수추출물 (CUI-0)이 Peyer’s patch를 매개로 한 가장 높은 장관면역 활성을 나타내었으며 다시마(L. japonica), 둥글레(P. japonicum), 탱자(P. trifoliata)의 냉수추출물과 구기자(L. chinense) 및 치자(G. jasminoides)의 열수추출물을 제외한 나머지 식물의 용매추출물은 거의 활성을 가지지 못하였다. CUI-0는 MeOH-가용성 획분(CUI-1), MeOH-불가용성이면서 EtOH-가용성 획분(CUI-2)과 조다당 획분(CUI-3)으로 분획되었다. 이러한 획분들 중 CUI-3은 Peyer’s patch 세포를 매개로 하는 골수세포 증식의 자극활성이 가장 높았으며 arabinose, galacturonic acid, galactose, glucose, glucuronic acid와 rhmanose(molar ratio; 1.00:0.53:0.45:0.28:0.28:0.19) 등을 주요 구성당으로 함유하고 있었으며 소량의 단백질(9.4%)도 구성물질로 포함된 활성 획분임이 밝혀졌다. CUI-3의 장관면역 활성은 pronase 및 periodate 처리에 의해 감소되었으며 특히 periodate 산화는 CUI-3의 활성에 심각한 영향을 끼치는 것을 알 수 있었다. 활성 획분으로서 당 함량이 높은 CUI-3IIb-3-2는 DEAE-Sepharose FF, Sepharose CL-6B 및 Sephacryl S-200에 의해 귤피 냉수추출물의 조다당 획분으로부터 정제되었으며 HPLC에 의해 분자량 약 18,000 Da의 단일 peak임을 확인하였다. CUI-3IIb-3-2는 주로 arabinose, galactose, rhamnose, galacturonic acid와 glucuronic acid(molar ratio; 1.00:0.54:0.28:1.45:0.63) 등의 구성당으로 구성되어져 있었으며 소량의 단백질(3.2%)이 함유되어진 물질로 구성되어 있었다. 한편 CUI- 3IIb-3-2는 Peyer’s patch를 경유하였을 때만 골수세포 증식활성을 나타내었으며 활성물질 자체가 직접 골수세포 증식활성에 관여하지는 않는것으로 확인되었다. 이러한 결과로부터 귤피의 장관면역 활성은 측쇄에 arabinose와 galactose 등의 중성당이 결합된 polygalacturonan 구조를 갖는 펙틴계통의 다당류에 기인하고 있음을 보여주었다.

Keywords

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