Antioxidant and tyrosinase inhibitory activity of white beech mushroom (Hypsizygus marmoreus) extracts

흰색 느티만가닥버섯 추출물의 항산화 활성 및 tyrosinase 저해 효과

Abstract

The objective of this study was to evaluate antioxidant effect and tyrosinase inhibitory activity of white beech mushroom (Hypsizygus marmoreus) extracts. The white beech mushroom was extracted into hot water and methanol. Total polyphenol content was highest in the hot water extract ($8.4{\pm}3.27mg\;GAE/g$) compared to the methanol extract ($7.3{\pm}2.85mg\;GAE/g$). The flavonoids contents in hot water and methanol extracts were $3.8{\pm}3.81ug/mg$ and $2.5{\pm}1.95ug/mg$, respectively. The tyrosinase inhibitory activity of extract was increased in a dose dependent manner and tyrosinase inhibitory activity of extract (hot water extract, 69.72%; methanol extract, 52.67% at 40 mg/ml) was lower than those of positive control 2% arbutin (96%). The DPPH radical scavenging activity of the hot water and methanol extract was 80% and 74%, respectively. Hot water extract ($63.34{\pm}1.00uM\;TE/g$) were more effective in ORAC (oxygen radical absorbance capacity) value than methanol extract ($46.33{\pm}0.48uM\;TE/g$). The toxicity of hot water and methanol extracts was investigated using WST-1 (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate) assay on the B16BL6 melanoma cells.

본 연구에서는 기능성 식의약품 및 화장품 소재로써 흰색 느티만가닥버섯(Hypsizygus marmoreus)의 이용 가능성을 조사하기 위해서 흰색 느티만가닥버섯 열수 추출물과 메탄올 추출물의 항산화 활성 및 tyrosinase 저해 효과를 비교하였다. 열수 추출물과 메탄올 추출물의 총 폴리페놀 함량은 각각 $8.4{\pm}3.27mg\;GAE/g$과 $7.3{\pm}2.85mg\;GAE/g$이었고 플라보노이드 함량은 각각 $4.8{\pm}3.81ug/mg$과 $2.5{\pm}1.95ug/mg$이었으며 총 폴리페놀과 플라보노이드 함량 모두 메탄올 추출물보다 열수 추출물에서 높게 나타났다. Tyrosinase 저해 활성은 추출물의 농도에 따라 증가하는 경향을 나타내었으나 양성 대조구로 사용한 2% 알부틴(arbutin)비해 40 mg/ml의 높은 농도에서도 열수 추출물은 69.72%, 메탄올 추출물은 52.67%의 낮은 저해 활성을 나타내었다. 항산화 활성은 DPPH에 의한 라디칼소거 활성을 측정하여 확인하였으며 열수 추출물과 메탄올 추출물의 DPPH 라디칼 소거능은 40 mg/ml의 농도에서도 각각 80%와 74%로 낮게 나타났다. 추출물의 세포독성은 WST-1 assay (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulphonate)를 이용하여 추출물의 처리농도에 따른 B16BL6 melanoma cell의 세포생존율로 확인하였으며 열수 추출물과 메탄올 추출물을 각각 0-40 mg/ml의 농도로 처리하였을 때 B16BL6 melanoma cell은 90% 이상의 생존율을 나타내었으므로 열수 추출물과 메탄올 추출을 모두 B16BL6 melanoma cell에 독성을 나타내지 않는 것으로 판단된다.

Fig. 1. Total polyphenolic and flavonoids contents of hot water and methanol extract of white beech mushroom (Hypsizygus marmoreus). Values are expressed as mean±SD (n=3), Values with different superscript letters are significantly different at p<0.05 by Duncan’s multiple range test.

Fig. 2. Tyrosinase inhibitory activity of hot water and methanol extract of white beech mushroom (Hypsizygus marmoreus). Values are expressed as mean±SD (n=3), Values with different superscript letters are significantly different at p<0.05 by Duncan’s multiple range test.

Fig. 3. DPPH-radical scavenging activity of hot water and methanol extract of white beech mushroom (Hypsizygus marmoreus). Values are expressed as mean±SD (n=3), Values with different superscript letters are significantly different at p<0.05 by Duncan’s multiple range test.

Fig. 4. Oxygen radical absorbance capacity (ORAC) of hot water and methanol extract of white beech mushroom (Hypsizygus marmoreus). Values are expressed as mean±SD (n=5), values with different superscript letters are significantly different at p<0.05 by Duncan’s multiple range test.

Fig. 5. Effects of white beech mushroom (Hypsizygus marmoreus) extracts on cell proliferation in B16BL6 mouse melanoma cell. The B16BL6 mouse melanoma cell was incubated for 24 hr in DMEM media with different concentration of hot water and methanol extract of white beech mushroom (Hypsizygus marmoreus). The cell proliferation was determined using WST-1 assay. Values are expressed as mean±SD (n=3), Values with different superscript letters are significantly different at p<0.05 by Duncan’s multiple range test.