Extraction, purification and properties of anti-complementary polysaccharide from Arecae Pericarpium

대복피로부터 항보체 활성다당의 추출, 정제 및 그 특성

  • Published : 1992.08.30

Abstract

To examine the characteristics of anti-complementary compounds from Arecae Pericarpium (the pericarps of Areca catechu) which showed the highest activity during our screening procedures, the extraction and purification were performed. AC-1 fraction from Arecae Pericarpium was purified by hot water extraction, methanol reflux, ethanol precipitation, dialysis and lyophilization. This compound had total sugar 48.2%, uronic acid 14.6% and protein 36.8%. Rhamnose, arabinose, mannose and galactose were found in sugar components. By cetavlon (cetyltrimethylammonium bromide) treatment AC-1 was fractionated to AC-2, AC-3 and AC-4. Among them, AC-2 showed the highest activity and yield. By periodate oxidation, AC-2 was deactivated, but had no change in activity by pronase digestion. Moreover active fractions, AC-2-IIIa and AC-2-IIIc isolated from AC-2 by two successive column chromatography using DEAE-Toyopearl $650C(Cl^-form)$ and Sephadex G-100. AC-2-IIIa was mainly made up of rhamnose, mannose, galactose and glucose, and AC-2-IIIc, mannose, galactose and glucose. These both polysaccharides were identified as homogeneous by gel filtration of Sepharose CL-4B and electrophoresis, and molecular weights of them were 120,000 and 15,000, respectively.

선별과정 중 가장 높은 항보체 활성을 보였던 대복피를 대상으로 활성물질의 특성을 조사하기 위하여 추출 및 정제를 행하였다. 대복피를 열수추출, methanol 환류 및 ethanol 침전을 행한 후, 투석 및 동결건조를 거쳐 활성획분 AC-1을 얻었는 바, 이들은 총당 48.2%, 산성당 14.6%, 단백질 36.8%로 구성되어 있었고 주구성당으로 rhamnose, arabinose, mannose 및 galactose를 함유하고 있었다. 이들은 다시 cetavlon(cetyltrimethylammonium bromide) 처리에 의해 AC-2, AC-3, AC-4로 분획되었으며 특히 AC-2의 경우 활성과 수율면에서 우수 하였다. 이들은 periodate 산화에 의해 활성의 감소를 일으킨 반면 pronase 소화에 의해서는 활성의 변화가 없는 특성을 보여 주었다. 더우기 AC-2는 DEAE-Toyopearl 650C($Cl^-$형) 및 Sephadex G-100을 이용한 연속적인 column chromatography를 통하여 활성이 우수한 AC-2-IIIa와 AC-2-IIIc로 분획되었으며 전자의 경우 rhamnose, mannose, galactose 및 glucose가, 후자의 경우 mannose, galactose 및 glucose가 주구성당이었다. 이들 활성다당들은 gel 여과 및 전기영동 결과, 순수한 물질임이 확인되었으며 이들의 분자량은 각각 120,000 및 15,000 이었다.

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