Applied Biological Chemistry

The Korean Society for Applied Biological Chemistry (한국응용생명화학회)
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Characteristics and purification of proteoglycan from Phellinus igniarius

Phellinus igniarius로부터 분리한 단백다당류의 분리 및 특성

  • Kim, Seon-Hee (Dept. of food science and technology, Yeungnam University) ;
  • Jung, In-Chang (Dept. of Hotel culinary art, Tonghae Junior College) ;
  • Kwon, Yong-Il (Hyupsungnongsan Co., Ltd) ;
  • Kim, So-Yeun (Dept. of food science and technology, Yeungnam University) ;
  • Lee, Jong-Suk (Dept. of food science and technology, Yeungnam University) ;
  • Lee, Hang-Woo (Dept. of Biology, Yeungnam University) ;
  • Lee, Jae-Sung (Dept. of food science and technology, Yeungnam University)
  • Published : 2000.02.29
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Abstract

The proteoglycan, intracellular and extracellular, extracted from the liquid culture of Phellinus igniarius were purified and characterized. The mycelial productivity was proved to be better in shaking culture compared to standing culture. The productivity of intracellular proteoglycan of Phellinus igniarius appeared to be similar in two culturing methods. The standing culture of Phellinus igniarius produced 6 times as much extracellular proteoglycan compared to shaking culture. The proteoglycan were purified to a single peak by ion exchange chromatography(DEAE-cellulose) followed by gel filtration(Sepharose 2B). PIEPDG contained 79.0% total sugar and 7.2% protein. PIEPAG contained 56.7% total sugar and 40.8% protein. PIIPDG contained 64.8% total sugar and 17.4% protein. PIIPAG contained 56.9% total sugar and 41.5%n protein. The molecular weights of all the fractions were estimated to be above 100,000, from 134KDa of PIEPDG to 560 KDa of PIEPAG. The results of sugar analysis by HPLC showed that PIEPDG contains glucose only. The sugar part of PIIPDG and PIIPAG were consisted of glucose and inositol. The PIEPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

Phellinus igniarius의 배양방법별 균사체 및 단백다당류 생산수율비교 실험에서는 균사체내 다당류의 경우 모두 진탕배양이 효과적이었으나 균사체외 다당류의 경우 다당류 생산에서는 정치배양이 효과적이었다. 다당류의 정제는 조단백다당류를 이용하여 DEAE-cellulose column에 의한 1차 정제를 행하였고, 최종으로 Sepharose 2B를 이용한 2차 정제를 실시하여 최종적으로 균사체내 단백다당류의 탈이온수 분획물(PIIPDG)과 알칼리분획물(PIIPAG), 균사체외 단백다당류의 탈이온수 분획물(PIEPDG) 알칼리 분획물(PIEPAG)을 얻었다. 이때의 정제 수율은 1차 정제에서 40%의 회수율을 나타내었으며 2차 정제에서는 $38{\sim}61%$의 높은 회수율을 보였다. PIEPDG는 총당 79.0%, 총단백질 7.2%, PIEPAG는 총당 56.7%, 총단백질 40.8%, PIIPDG는 총당 64.8%, 총단백질 17.4%, PIIPAG는 총당 56.9%, 총단백질 41.5%으로 측정되었다. 각 단백다당류의 분자량은 PIEPDG 166KDa에서 PIEPAG 565KDa까지 모두 10만이 넘는 거대분자로 나타났다. 각 분획물의 단당류 조성을 볼때, PIEPDG는 glucose, PIIPDG와 PIIPAG는 glucose, inositol, PIEPAG는 glucose, rfructose, inositol이 검출되었다.

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