[논문]잎새버섯 균사체를 이용한 감초추출발효물의 플라보노이드 생성과 항염 활성 연구

배준태; 송민현; 김진화; 이근수; 표형배

doi:10.15230/scsk.2012.38.4.327

[국내논문] 잎새버섯 균사체를 이용한 감초추출발효물의 플라보노이드 생성과 항염 활성 연구
Production of Flavonoid Compounds and Anti-inflammatory Property of Fermented Licorice Extract with the Basidiomycete Grifola frondosa HB0071 원문보기

大韓化粧品學會誌 = Journal of the society of cosmetic scientists of Korea, v.38 no.4, 2012년, pp.327 - 338

배준태 (한불화장품(주) 기술연구소) , 송민현 (한불화장품(주) 기술연구소) , 김진화 (한불화장품(주) 기술연구소) , 이근수 (한불화장품(주) 기술연구소) , 표형배 (한불화장품(주) 기술연구소)

초록
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리퀴리티게닌과 이소리퀴리티게닌은 감초의 주요 플라보노이드 성분이다. 이들 플라보노이드는 수용성 감초 추출물과 ${\beta}$-glucosidase를 생성하는 잎새버섯 HB0071 균사체 발효배양을 통하여 생산하였다. 감초추출물 내 리퀴리티게닌과 이소리퀴리티게닌은 잎새버섯 발효배양 동안 현저히 증가하였다. 이 균주의 ${\beta}$-glucosidase의 활성은 배양 96시간을 기준으로 최고 91.5 mU/mL로 확인되었으며, 감초추출발효물로부터 생성된 리퀴리티게닌과 이소리퀴리티게닌의 함량은 HPLC 분석을 통하여 최대 $568.5{\mu}g/mL$과 $89.6{\mu}g/mL$로 확인되었다. 본 연구에서는 감초추출물의 잎새버섯 발효 전 후의 시료가 처리된 각질형성세포를 이용하여 자외선 UVB에 조사로 발현된 염증유발인자(COX-2)와 사이토카인(IL-$1{\beta}$, IL-6) 모두 감초추출발효물(FLEx)에서 농도의존적으로 발현이 억제되는 것을 확인하였다. 결론적으로 리퀴리티게닌과 이소리퀴리티게닌의 함량이 증가된 감초추출발효물은 자외선으로부터 손상된 피부 염증반응을 완화시켜줄 것으로 사료된다.

Abstract ▼ AI-Helper

Liquiritigenin and isoliquiritigenin are the major flavonoids present in licorice. These flavonoid compounds were prepared by submerged culture of Grifola frondosa (G. frondosa) HB0071 mycelia producing ${\beta}$-glucosidase in the aqueous extract of licorice. The contents of liquiritigenin and isoliquiritigenin were increased during the fermentation. This fungus produced a high ${\beta}$-glucosidase (activity of 91.5 mU/mL), thereby achieving high amounts of liquiritigenin and isoliquiritigenin ($568.5{\mu}g/mL$ and $89.6{\mu}g/mL$), respectively at 96 h. A reversed- phase high-performance liquid chromatography method was established for simultaneous determination of liquiritigenin and isoliquiritigenin in fermented licorice extract (FLEx). The anti-inflammatory activities were investigated by licorice extract (LEx) before and after fermentation with G. frondosa HB0071. The treatment of UVB-irradiated HaCaT keratinocytes with FLEx resulted in a dose-dependent decrease in the expression level of cyclooxygenase-2 (COX-2) mRNA. Furthermore, FLEx dose-dependently decreased mRNA of the pro-inflammatory cytokines of IL-$1{\beta}$ and IL-6 in UVB-irradiated HaCaT cells. These results suggest that FLEx may mitigate the effects of skin inflammation by reducing UVB-induced adverse skin reactions.

Keyword

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문제 정의

The objective of the work was to develop liquiritigenin and isoliquiritigenin production from a submerged culture of G. frondosa HB0071 using licorice extract as substrate. The effects of fermented licorice extract with G.

제안 방법

In order to isolate the region of liquiritigenin and isoliquiritigenin by flavonoid compounds with much more retention time, the elution was carried out with gradient mobile phase (A) aqueous acetic acid (6 mM) and (B) methanol, starting from 80 % A to 40 % A for 0 ∼ 110 min, 40 ∼ 0 % A for 10 min and 80 % A for 10 min.
5 mU/mL, respectively. To examine whether liquiritigenin and isoliquiritigenin in licorice extract were increase by the fermentation by G. frondosa, the detected major peaks in licorice extract and fermented licorice extract were analyzed by HPLC. Figure 3 shows the HPLC chromatogram at 327 nm of licorice extract (Figure 3A) fermented with G.
To investigate whether LEx and FLEx inhibit UVB-induced COX-2 expression, HaCaT cells were exposed to UVB (40 mJ/cm²). Thus, in the present study the effects of LEx and FLEx upon UVB irradiation were examined by measuring the steady-state COX-2 mRNA level in relation to GAPDH mRNA levels.

대상 데이터

All experiments were performed in triplicate. Data were presented as meanstandard error (SE).

데이터처리

Statistical comparison was conducted using Student’s t-test after ANOVA.

성능/효과

In this study, there were a greater amount of liquiritin and isoliquiritin than liquiritigenin and isoliquiritigenin in aqueous licorice extracts due to the poor water-solubility characteristics of flavonoid aglycones (data not shown). Interestingly, however, liquiritigenin and isoliquiritigenin contents increased after fermentation (Table 1).
In summary, our results demonstrated that fermented licorice extract with G. frondosa HB0071 was very effective for the improvement of flavonoid aglycones (liquiritigenin and isoliquiritigenin) contents and inhibitory effects on UVB-induced inflammation in HaCaT keratinocytes. We also investigated that exopolysaccharide (EPS) was produced from submerged culture of aqueous licorice extract with G.

후속연구

frondosa HB0071 mycelia, is a potential candidate to reduce UV-induced skin inflammation. In the future, combinations of other natural compounds may be considered for more efficient photoprotection, and further studies for elucidating overall biological functions of FLEx in vivo should be performed.

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