Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Analysis of ${\beta}-(1{\to}3)(1{\to}6)-Glucan$ Produced by Aureobasidium pullulans IMS-822

Aureobasidium pullulans IMS-822가 생산하는 ${\beta}-(1{\to}3)(1{\to}6)-Glucan$의 특성 분석

  • Lee, Seog-June (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Ahn, Keug-Hyun (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Chan-Sun (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Yoon, Byung-Dae (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Min-Soo (Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • 이석준 (한국생명공학연구원 생물산업기술연구센터) ;
  • 안극현 (한국생명공학연구원 생물산업기술연구센터) ;
  • 박찬선 (한국생명공학연구원 생물산업기술연구센터) ;
  • 윤병대 (한국생명공학연구원 생물산업기술연구센터) ;
  • 김민수 (한국생명공학연구원 생물산업기술연구센터)
  • Published : 2009.03.31
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Abstract

The exo-polysaccharide producing microorganism, Aureobasidium pullulans IMS-822, was isolated and identified from soil. The viscosity-average molecular weight of exo-polysaccharide was calculated as $8.9{\times}10^5$ by Mark-Houwink equation. The sugar component of exo-polysaccharide was determined as glucose by HPLC analysis. The IR spectra indicated that the exo-polysaccharide has an absorption peak at 890 $cm^{-1}$ for the ${\beta}-configuration$ of D-glucan. The $^{13}C$ NMR signal at ${\delta}$ 86.62 ppm arose from the substituted C-3 of glucose. The signal at ${\delta}$ 72.11 ppm was assigned to C-6 of branched ${\beta}-(1{\to}3)-D-glucosyl$ residues. Viscosity and Congo red reaction indicated that {\beta}-(1{\to}3)(1{\to}6)-glucan$ produced by A. pullulans IMS-822 has a highly ordered hydrogen-bond dependent conformation in aqueous solution, which collapses in strong alkaline solution.

국내 토양시료로부터 점액성 고분자물질을 생산하는 효모 균주를 분리 및 동정하여 Aureobasidium pullulans IMS-822라 명명하였다. A. pullulans IMS-822가 생산하는 외분비성 폴리머를 정제 및 동결 건조하여 구조분석을 수행하였다. HPLC를 이용한 구성당 성분분석에서 포도당 단일성분으로 구성되어 있음을 확인하였다. FT-IR을 이용한 구조분석에서는 890 $cm^{-1}$ 부근에서의 ${\beta}-configuration$을 확인하였으며, 3300 $cm^{-1}$ 부근으로 shift 된 완만한 -OH stretching은 폴리머를 구성하고 있는 분자 사이에 강한 수소결합이 작용하고 있는 것으로 추측되었다. $^{13}C-NMR$ 분석결과는 A. pullulans IMS-822가 생산하는 외분비형 다당류인 (1,3)-linked ${\beta}-D-glucosyl$ unit의 6개의 탄소, ${\delta}$ 105.05 ppm (C1), 75.82 ppm (C2), 86.62 ppm (C3), 70.61 ppm (C4), 78.13 ppm (C5), 63.22 ppm (C6) 및 ${\beta}-(1,3)D-glucosyl$ 잔기에 분기사슬 되어 있는 ${\delta}$ 72.11 ppm의 C-6의 signal을 확인할 수 있었다. A. pullulans IMS-822가 생산하는 ${\beta}-(1{\to}3)(1{\to}6)-glucan$의 알칼리 수용액(0.5%, w/v)에 24.0 ${\mu}M$의 Congo red 첨가하여 30분간 반응시킨 후 $400{\sim}700$ nm의 범위에서 최대흡수파장의 변화를 조사한 결과 NaOH의 농도가 증가함에 따라 최대흡수파장은 장파장으로 이동하여 498 nm에서 515 nm까지 이동하였다. 최대흡수파장은 0.4 M NaOH 농도에서 점차로 감소하여 506 nm 부근에서 안정화되는 것으로 나타남으로써 순차적 구조를 가지는 것으로 확인되었다.

Keywords

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