Microbiology and Biotechnology Letters (한국미생물·생명공학회지)

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Conversion of Ginsenoside $Rb_1$ by Ginseng Soil Bacterium Cellulosimicrobium sp. Gsoil 235 According to Various Culture Broths

인삼 토양 미생물 Cellulosimicrobium sp. Gsoil 235의 배지조성에 따른 Ginsenoside $Rb_1$ 전환

  • Na, Ju-Ryun (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Yu-Jin (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Se-Hwa (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Ho-Bin (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Shim, Ju-Sun (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Kim, Se-Young (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University) ;
  • Yang, Deok-Chun (Korean Ginseng Center Most Valuable Products and Ginseng Genetic Resource Bank, Kyung Hee University)
  • 나주련 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김유진 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김세화 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김호빈 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 심주선 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 김세영 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행) ;
  • 양덕춘 (경희대학교 고려인삼명품화사업단 및 인삼유전자원소재은행)
  • Published : 2009.03.28
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Abstract

Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having $\beta$-glucosidase (to transform ginsenoside $Rb_1$) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-$886^T$. The strain, Gsoil 235, could transform ginsenoside $Rb_1$ into Rd, $Rg_3$ and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for, $\beta$-glucosidase was nutrient broth (In 48 hours, almost ginsenoside $Rb_1$ could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).

인삼 근권에 존재하는 토양 미생물중 esculin agar법을 이용하여 $\beta$-glucosidase를 생산하는 균주를 분리하고, 다시 ginsenoside $Rb_1$을 선택적으로 분해하는 균주 Gsoil 235를 선발 및 동정하였다. 16S rRNA 염기서열을 sequencing한 후, genebank에서 가장 가까운 type strain을 결정하여 유연 관계를 분석한 결과 Cellulosimicrobium 속의 funkei ATCC BAA-$886^T$(AY501364)와 99.7% 일치하는 균주임을 확인하였다. TSB, LB, NB등 3종류의 배지에서 균의 생장은 접종 후 12-24 시간에서 가장 잘 자라며, TSB>LB>NB의 순으로 잘 자라는 것을 알 수 있었다. ginsenoside $Rb_1$과 8, 24, 48시간 동안 반응시킨 후 TLC로 분석한 결과 NB>LB>TSB순으로 $Rb_1$ 분해 활성이 뛰어나 배지의 생장과 대조적인 결과를 얻었다. 반응시간이 증가할수록 Rd를 포함한 minor ginsenoside의 생성이 증가하였으며, 특히 다른 배지에 비해 균주 생장속도가 상대적으로 낮은 NB는 48시간 후 $Rb_1$을 거의 분해하여 강한 효소 활성을 확인할 수 있었다.

Keywords

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