The Korean Journal of Mycology (한국균학회지)

The Korean Society of Mycology (한국균학회)
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Isolation and purification of protein-bound polysaccharides from the sawdust mycelia of Agrocybe cylindracea

톱밥배양한 버들송이의 균사체로부터 단백다당류의 분리 및 정제

  • Ha, Hyo-Cheol (Department of Food Science and Technology, Yeungnam University) ;
  • Park, Shin (Department of Agricultural Chemistry, Taegu University) ;
  • Park, Kyung-Sook (Department of Food Science and Technology, Yeungnam University) ;
  • Lee, Chun-Woo (Department of Food Science and Technology, Yeungnam University) ;
  • Jung, In-Chang (Department of Food Science and Technology, Yeungnam University) ;
  • Kim, Seon-Hee (Department of Food Science and Technology, Yeungnam University) ;
  • Kwon, Yong-Il (Department of Food Science and Technology, Yeungnam University) ;
  • Lee, Jae-Sung (Department of Food Science and Technology, Yeungnam University)
  • 하효철 (영남대학교 식품가공학과) ;
  • 박신 (대구대학교 농화학과) ;
  • 박경숙 (영남대학교 식품가공학과) ;
  • 이춘우 (영남대학교 식품가공학과) ;
  • 정인창 (영남대학교 식품가공학과) ;
  • 김선희 (영남대학교 식품가공학과) ;
  • 권용일 (영남대학교 식품가공학과) ;
  • 이재성 (영남대학교 식품가공학과)
  • Published : 1995.06.30
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Abstract

The characteristics of protein-bound polysaccharides (PBP) which were isolated and purified from the sawdust mycelia of Agrocybe cylindracea were investigated. The yield of crude protein-bound polysaccharides (Fr.CB) extracted with boiling water and precipitated with 95% ethanol, was 0.74% based on the original sawdust mycelia. The Fr.CB was purified by the membrane filtration, ion exchange chromatography and gel filtration. The Fr.B fraction of which the molecular weight is over 300 KDa, was isolated from the Fr.CB using membrane filtration, and the yield was 38.6% based on the Fr.CB. This result indicates that high molecular protein-bound polysaccharides are the dominent components of the Fr.CB. Two fractions (Fr.B-1, Fr.B-2) were also isolated from the Fr.B using ion exchange chromatography, and the yields were 17.3% (Fr.B-1) and 10.3% (Fr. B-2), respectively. The Fr.B-1 was concentrated and gel-filtrated, and the single peak, thought to be nearly pure protein-bound polysaccharides, was obtained. The yield of final fraction $(Fr.B-1-{\beta})$ was 42.5% based on the Fr.B-1. The molecular weight of $Fr.B-1-{\beta}$ was nearly 710 KDa. The monosaccharides' composition of $Fr.B-1-{\beta}$ was analized by HPLC, and glucose was the dominent component, and fucose and galactose were also detected. The result of amino acid analysis was that glutamic acid and analysis were detected to a significant level, and cysteine was not detected.

Agrocybe cylindracea의 톱밥배양 균사체로부터 항암효과가 있는 것으로 알려진 단백다당류를 분리, 정제하였으며 그 특성을 조사하였다. Agrocybe cylindracea 균사체로부터 열수추출한 조단백다당류(Fr.CB)의 수율은 ethanol 농도가 95%일 때 2.974 g으로서 원재료인 톱밥균사체를 기준으로 0.74%였다. 이 조단백다당류(Fr.CB)를 박막여과, ion exchange chromatography, 그리고 gel filtration에 의해 정제하였다. 조단백다당류 Fr.CB를 박막여과하여 분획한 결과 분자량 30만 이상의 분획(Fr.B)이 38.6%를 차지하여 고분자 단백다당류가 주성분임을 알 수 있었다. Fr.B를 ion exchange chromatography로 분리한 결과 2개의 분획이 17.4%(Fr.B-1), 10.3%(Fr.B-2)의 수율로 얻어졌다. 이들 분획을 농축한 다음 gel filtration한 결과 거의 순수한 단일 단백다당류의 peak를 얻을 수 있었다. Fr.B-1을 분리하여 얻은 $Fr.B-1-{\beta}$ 분획의 수율은 Fr.B-1을 기준으로 42.5%였다. 항암효과의 가능성이 가장 높은 것으로 판단되는 최종 정제된 분획인 $Fr.B-1-{\beta}$의 분자량은 710 KDa 부근이었으며 단당류의 조성을 HPLC로 분석한 결과, glucose가 주성분이었고 그외 fucose, galactose도 검출되었다. 또한 아미노산 조성을 분석한 결과 glutamic acid, alanine이 비교적 많이 검출되었고 cysteine은 검출되지 않았다.

Keywords

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