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Isolation of Stenotrophomonas rhizopilae Strain GFC09 with Ginsenoside Converting Activity and Anti-wrinkle Effects of Converted Ginsenosides

사포닌 전환 활성 Stenotrophomonas rhizopilae Strain GFC09 균주의 분리 동정 및 전환 사포닌의 주름 개선 효과

  • 민진우 ((주)지에프씨 생명과학연구원) ;
  • 김혜진 (네오바이오연구소 기업부설연구소) ;
  • 주광식 (네오바이오연구소 기업부설연구소) ;
  • 강희철 ((주)지에프씨 생명과학연구원)
  • Received : 2015.09.21
  • Accepted : 2015.09.27
  • Published : 2015.12.30

Abstract

Ginsenosides (ginseng saponin) as the one of important pharmaceutical compounds of ginseng and is responsible for the pharmacological and biological activities. These ginsenoside produces diverse small molecules ginsenoside which have more pharmacological activities including anti-wrinkle, anti-cancer and anti-oxidant effects. In the present study, we isolated bacteria using esculin agar, to produce ${\beta}$-glucosidase, and we focused on the bio-transformation of ginsenoside. Phylogenetic tree analysis was performed by comparing the 16S rRNA sequences; we identified the strain as Stenotrophomonas rhizopilae strain GFC09. In order to determine the optimal conditions for enzyme activity, the crude enzyme was incubated with 1 mM ginsenoside $Rb_1$. Bioconversion of ginsenoside $Rb_1$ were analyzed using TLC and HPLC. The crude enzyme hydrolyzed the ginsenoside $Rb_1$ along the following pathway: LB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$ into compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. The structure of the hydrolyzed metabolites were identified by NMR. The activity screening tests showed that the conversion product induced the production of type I procollagen in a dose-dependent manner. These results suggested that hydrolyzed ginseng product containing the ginsenoside $F_2$ and compound K could be useful as an active ingredient for wrinkle-care cosmetics.

진세노사이드(인삼 사포닌)는 인삼의 대표적 약리성분 중의 하나로 생물학적 활성을 가진 배당체 화합물이다. 이들 사포닌은 가수분해 되어 저분자화 되었을 때, 항주름 및 항산화, 항암 등에 높은 약리효능효과를 나타낸다. 본 연구에서는 인삼 esculin 배지를 활용하여 ${\beta}$-glucosidase 활성을 가진 균주를 분리하였고 인삼 사포닌 전환을 미생물을 이용하여 수행하였다. 본 균주들을 16S rRNA sequencing을 통하여 동정하여 본 결과 Stenotrophomonas rhizopilae strain GFC09로 확인되였다. 균주의 최적 활성 조건을 결정하기 위해 조효소 1 mM와 인삼사포닌 $Rb_1$과 함께 배양한 후 생물학적 전환을 TLC, HPLC를 사용하여 확인하였다. 조효소에 의한 인삼 사포닌 $Rb_1$의 전환 경로는 다음과 같다. LB: RbNeobio R&D center, Gyeonggi-do 16954, Korea${\rightarrow}$Rd${\rightarrow}$FNeobio R&D center, Gyeonggi-do 16954, Korea${\rightarrow}$compound K, TSB: $Rb_1{\rightarrow}Rd{\rightarrow}F_2$. 가수분해된 생성된 물질은 NMR로 구조 동정하였다. 전환 산물의 효능 분석결과, 콜라겐 생성을 농도 의존적으로 증가시키는 것이 관찰되었다. 이에 본 연구에서는 ginsenoside $F_2$와 compound K 함유 인삼 전환 산물의 주름 개선 소재로서 활용가능성을 확인하였다.

Keywords

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