[논문]Aspergillus usamii KCTC 6954에 의한 ginsenoside Rb1로 부터 의약용 소재인 compound K로의 생물학적 전환

조미나; 정지은; 윤현주; 장경훈; 지희숙; 김기태; 백현동

doi:10.4014/kjmb.1407.07010

Aspergillus usamii KCTC 6954에 의한 ginsenoside Rb1로 부터 의약용 소재인 compound K로의 생물학적 전환
Bioconversion of Ginsenoside Rb1 to the Pharmaceutical Ginsenoside Compound K using Aspergillus usamii KCTC 6954 원문보기

한국미생물·생명공학회지 = Korean journal of microbiology and biotechnology, v.42 no.4, 2014년, pp.347 - 353

조미나 (건국대학교 축산식품공학과) , 정지은 (건국대학교 축산식품공학과) , 윤현주 (건국대학교 축산식품공학과) , 장경훈 (CJ 제일제당 식품연구소) , 지희숙 ((주)일화 중앙연구소) , 김기태 (건국대학교 생명분자정보학센터) , 백현동 (건국대학교 축산식품공학과)

초록
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본 연구는 인삼의 주요성분인 ginsenoside Rb1으로부터 보다 높은 생리기능성을 갖는 것으로 알려져 있는 compound K를 생산하기 위하여 Aspergillus usamii KCTC 6954에서 유래된 ${\beta}$-glucosidase를 사용하여 생물전환을 실시하였다. 15일 동안의 배양 중, 효소활 성 측정은 ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside를 기질로 하여 분해 생성되는 ${\rho}$-nitrophenol (${\rho}NP$)을 비색계로 측정함으로써 실시되었다. 그 결과로서, 균주의 성장 속도는 접종 후 6일 후 최대로 나타났으며 이때의 ${\beta}$-glucosidas 활성도는 $175.93{\mu}M\;ml^{-1}min^{-1}$로 나타났다. 또 한 효소 반응의 최적 조건은 pH 6.0 이내에서는 $60^{\circ}C$인 것으로 나타났다. 배양 중 ginsenosides 분석 결과, 배양 9일 후에는 Rb1는 Rd 로 전환되고 15 days 후에는 compound K로 순차적으로 전환되는 것으로 나타났다. 효소반응에 있어서는 Rb1는 1시간 이내에 ginsenoside Rd로 전환되었고 8시간 이후에 최종산물인 compound K가 측정되었다. 본 연구결과로부터 Rb1으로부터 주요 생물학적 전환 경로는 $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K로 나타났으며 이는 차후 Rd나 compound K와 같이 강한 생리기능성을 갖지만 자연에 미 량 존재하는 물질의 대량생산에 응용될 수 있을 것으로 기대된다.

Abstract ▼ AI-Helper

${\beta}$-Glucosidase from Aspergillus usamii KCTC 6954 was used to convert ginsenoside Rb1 to compound K, which has a high bio-functional activity. The enzymatic activities during culturing for 15 days were determined using ${\rho}$-nitrophenyl-${\beta}$-glucopyranoside. The growth rate of the strain and the enzymatic activity were maximized after 6 days (IU; $175.93{\mu}M\;ml^{-1}\;min^{-1}$). The activities were maximized at $60^{\circ}C$ in pH 6.0. During culturing, Rb1 was converted to Rd after 9 d and then finally converted to compound K at 15 d. In the enzymatic reaction, Rb1 was converted to the ginsenoside Rd within 1 h of reaction time and compound K could be detected after 8 h. As a result, this study demonstrates that $Rb1{\rightarrow}Rd{\rightarrow}F2{\rightarrow}$compound K is the main metabolic pathway catalyzed by ${\beta}$-glucosidase and that ${\beta}$-glucosidase is a feasible option for the development of specific bioconversion processes to obtain minor ginsenosides such as Rd and compound K.

주제어

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문제 정의

Therefore, the objective of this study is to investigate the enzymatic conversion of ginsenosides Rb1 to compound K by using enzymes, particularly β-glucosidase, produced by using Aspergillus usamii KCTC 6954 as basic information for industrial production.

대상 데이터

A. usamii KCTC 6954 was purchased from the Korean Collection for Type Cultures (KCTC; Daejeon, Korea). The fungus was grown on potato dextrose agar at 30℃ for 4 days and the stock culture was maintained at 4℃.
All experiments were performed in triplicate. The data was analyzed by using SPSS 18 (Chicago, IL, USA).

데이터처리

The mean values were determined by one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test (p < 0.05).

이론/모형

Enzyme activity of culture broth was evaluated by a colorimetric method using ρNPG as the substrate [19].

성능/효과

After 9 days of culturing, β-glucosidase activity was a little decreased, not significantly (p < 0.05), at this high level for the duration of the experiment.
In conclusion, the results of HPLC demonstrated that ginsenoside Rb1→Rd→F2→compound K (Fig. 5) is the main metabolic pathway catalyzed by A. usamii KCTC 6954 enzymes indicating that this result is similar to the pathway in Actinosynnema mirum [5].

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