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Isolation of Immunomodulatory Antitumor Active Polysaccharide (RGAP) from Red Ginseng By-Product and Its Physico-chemical Properties

홍삼추출잔사로부터 항암면역조절 활성을 보여주는 홍삼산성다당체(RGAP)의 분리 및 이화학적 특성

  • Published : 2003.07.01

Abstract

This study was carried out in order to develop the method for isolation of red ginseng acidic polysaccharide (RGAP) haying immunomodulating antitumor activity from red ginseng by-product. The red ginseng by-product was obtained from red ginseng residues produced in processing of red ginseng ethanol extract. The yield of RGAP isolated by ultrafiltration was 20.9%. The active substance (GFP) was purified by DEAE-sepharose column chromatography RGAP induced nitric oxide (NO) exhibited tumoricidal activities against P8l5 (mastocytoma) tumor cells. Acid-hydrolyzed RGAP fragments were shown four to five spots. These sopts showed the same R$_{f}$ values with sugars designated as rhamnose, glucose, glactose and glucuronic acid. Some physico-chemical properties of RGAP were investigated. pH and dry reduction content at 105$^{\circ}C$ were 4.74 and 4.72%, respectively. Crude protein, ash and Pb contents were 3.30%, 4.74% and 2.30 ppm. These results suggest that we will be able to produce RGAP from red ginseng by-product by ultrafiltration in a large scale.e.

홍삼으로부터 홍삼알콜 추출물을 제조하고 부산물로 생성된 홍삼추출잔사로부터 한외 여과를 이용하여 홍삼산성다당체(RGAP, red ginseng acidic polysaccharide)를 대량으로 분리할 수 있는 방법을 개발하였다. 본 방법으로 홍삼추출잔사로부터 분리된 RGAP의 수율은 20.9%이었고, RGAP로부터 지표성분인 활성성분(GFP)의 분리수율은 39.4%이었다. 분리된 RGAP는 macrophage에 의한 NO(nitric oxide)를 생성하여 암세포를 사멸시키는 것으로 나타났다. RGAP를 BALB/c 마우스에 복강투여한 후 분리된 복강 macrophage를 P815 암세포(mastocytoma)와 함께 배양하였을 때 배지내의 NO 농도는 대조군에서 4.3 $\mu\textrm{m}$이었으나, RGAP 100, 300 mg/kg 투여군에서는 각각 9.8 및 25.3$\mu\textrm{m}$로 증가하는 경향을 나타내었다. 아울러 암세포인 P8l5세포의 사멸율도 정상대조군일 경우 7.3%이었으나 RGAP 100, 300 mg/kg 투여군에서는 각각 22.7 및 31.8%로 증가하는 경향을 나타내었다. RGAP을 2M의 TFA로 10$0^{\circ}C$에서 1시간 가수분해한 후 당 표준품과 TLC로 비교한 결과 당 표준품과 Rf치 및 발색 색깔이 일치하는 4개의 spot, 즉 rhamnose(Rf 0.78, 노란색 ), glucose(Rf 0.61, 녹색), galactose(Rf 0.56, 녹색), glucuronic acid(Rf 0.36, 황갈색)를 관찰할 수 있었다. 가수분해된 RGAP 분해산물을 Gl(glucose)부터 G7(maltoheptaose)까지의 당류와 함께 TLC를 행한 결과 진한 색상을 나타내는 spot은 Rf 0.56~0.61로 G2(maltose, Rf 0.46) 이상의 단당류(monosaccharide)인 것으로 나타났다. RGAP의 일부 이화학적 특성을 조사한 결과 pH는 4.74, 건조감량은 4.72%, 조단백질 함량은 3.30%, 회분은 4.74%이었고 중금속인 Pb는 2.30 ppm이었다.

Keywords

References

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