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

  • 제45권1호
  • /
  • Pages.52-60
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  • 2016
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  • 1226-3311(pISSN)
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  • 2288-5978(eISSN)
한국식품영양과학회 (The Korean Society of Food Science and Nutrition)
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감잎으로부터 면역 활성 다당의 선택 분리를 위한 간편 방법

Convenient Method for Selective Isolation of Immuno-Stimulating Polysaccharides from Persimmon Leaves

  • 투고 : 2015.10.15
  • 심사 : 2015.11.27
  • 발행 : 2016.01.31
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초록

다당이 갖는 각종 생물 활성은 그 구조의 특성 및 분자량 분포에 의해 달라지므로 특정 다당을 분리하기 위한 정제과정은 다당 연구에 있어 필수적으로 요청된다. 본 연구에서는 서로 다른 특성을 소유한 다당을 분획하기 위한 간편하고 신속한 분리법을 개발하기 위해 한국산 감잎으로부터 조제한 다당 혼합물을 이용, 본 분리법을 최적화하였다. 감잎은 pectinase 처리 후 에탄올 침전법을 통해 조다당 획분인 PLE로 조제되었으며, PLE는 재차 농도별로 연속 희석된 에탄올 용액(EtOH : DIW=4:1, 2:1, 1.5:1, 1:1 및 0.5:1)을 이용하여 총 10개 획분(5개 침전물 획분: PLE-4~PLE-0.5, 5개 상등액 획분: PLE-4S~PLE-0.5S)으로 분획하였다. HPLC 분석 결과 PLE-4, PLE-2 획분은 저분자와 고분자 획분이 혼합된 다당이, PLE-1.5~0.5 획분에는 고분자 다당이 주로 검출되었다. 또한 PLE-4~PLE-1 획분은 구성당 분석 결과 RG(rhamnogalacturonan)-I과 RG-II 다당의 지표 구성당인 총 13종의 서로 다른 당으로 구성되어 있음이 확인되었으며, ${\beta}$-arabino-3,6-galactan 잔기를 함유하고 있는 것으로 확인되었다. 하지만 PLE-0.5 획분에서는 RG-II 및 ${\beta}$-arabino-3,6-galactan 잔기를 함유하고 있지 않았다. 한편 PLE-1.5S~PLE-1S 획분을 처리한 마우스 복강 대식세포에서는 농도 의존적인 IL-6의 생산 증가가 관찰된 반면, 저분자 다당으로 구성된 PLE-4S 및 PLE-2S 획분에서는 활성이 매우 낮음이 확인되었다. 이상의 결과로부터 본 분리 방법이 다양한 특성을 갖는 다당의 혼합물로부터 생물 활성을 갖는 RG류를 신속하고 간편하게 분리하는 데 있어 유용한 방법임을 확인할 수 있었다.

The biological activity of polysaccharide is greatly influenced by polysaccharide structure and molecular distribution. Here, we developed a rapid and convenient isolation method for fractionating polysaccharides with different characteristics and optimized it using a polysaccharide mixture from Korean persimmon leaves. A crude polysaccharide mixture, persimmon leaves-enzyme (PLE) fraction, was isolated from persimmon leaves digested with pectinase and ethanol precipitation. The PLE fraction was further fractionated with a serially diluted ethanol solution (ethanol : deionized water=4:1, 2:1, 1.5:1, 1:1, and 0.5:1) to produce 10 subfractions (five precipitate fractions labeled from PLE-4 to PLE-0.5 and five supernatant fractions labeled from PLE-4S to PLE-0.5S). HPLC analysis indicated that PLE-4 and -2 consisted of diverse polysaccharides, whereas PLE-1.5, -1, and -0.5 contained high molecular weight (MW) polysaccharides. The fractions from PLE-4 to PLE-1 were mostly composed of 13 different characteristic sugars in rhamnogalacturonan (RG) I and II, and the sugars contained an arabino-${\beta}$-3,6-galactan moiety. However, PLE-0.5 did not contain RG-II or ${\beta}$-arabino-3,6-galactan. Treatment of macrophages with fractions PLE-1.5S and PLE-1S led to a $10{\mu}g/mL$ increase in interleukin (IL)-6 production, whereas treatment with PLE-4S and PLE-2S fractions composed of low MW polysaccharides resulted in reduced levels of IL-6. These results indicate that this isolation method may be useful for the rapid and convenient fractionation of bioactive RGs from polysaccharide mixtures with various properties.

키워드

참고문헌

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피인용 문헌

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