• 대한화장품학회지
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• 제35권1호
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• pp.73-78
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• 2009

닭의장풀(Commelina communis L. Dayflower) 추출물의 미백효능을 확인하기 위하여 B16 melanoma 세포를 이용하여 melanin 생성에 관련된 다양한 실험을 실시하였다. 닭의장풀 추출물은 버섯 tyrosinase 활성 실험에서 저해 효과를 보이지 않았으나, melanin 생성 저해효과를 나타내었다. B16-F10 멜라노마 세포를 이용한 활성시험 결과에서, 닭의장풀 추출물은 1,000 ${\mu}g/mL$의 농도에서 약 32%의 멜라닌 생성을 억제하였으며, 세포내 티로시나제 활성 억제 능도 우수하여 닭의장풀 추출물 250 ${\mu}/mL$ 이상의 농도에서 50% 이상의 저해 효과를 보였다. 추출물의 생성 기작에 대한 영향을 조사한 결과, melanin 합성의 key protein 인 tyrosinase 발현의 우수한 저해 능력을 보였고, tyrosinase related protein-1 (TRP-1)의 발현 억제와 tyrosinase related protein-2 (TRP-2)의 N-glycosylatin 억제를 통해 melanin 합성이 억제되는 것으로 분석되었다. 따라서 닭의장풀 추출물은 melanin 합성에 필수적인 효소(tyrosinase, TRP-1)의 발현 저해 및 TRP-2의 N-glycosylation 억제를 통해 미백 효과를 나타내는 것으로 확인되었으며, 이에 따라 본 추출물은 melanin 합성의 효소 경로를 저해하는 미백 소재로 활용할 수 있을 것으로 사료된다.

• Bulletin of the Korean Chemical Society
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• 제28권5호
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• pp.758-762
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• 2007

A new alkyldithiocarbonate (xanthate), as sodium salts, C2H5OCS2Na, was synthesized by the reaction between CS2 with ethyl alcohol in the presence of NaOH. The new xanthate was characterized by 1H NMR, IR and elemental analysis. Then, the new synthesized compound was examined for functional study of cresolase activity of Mushroom Tyrosinase (MT) from a commercial source of Agricus bisporus in 10 mM phosphate buffer pH 6.8, at three temperatures of 10, 20 and 33℃ using UV spectrophotemetry. 4-[(4-methylphenyl)- azo]-phenol (MePAPh) was used as a synthetic substrate for the enzyme for cresolase reaction. The results show that ethyl xanthate can activate or inhibit the cresolase activity of mushroom tyrosinase depending to the concentration of ethyl xanthate. It was concluded that the enzyme has two distinct sites for ethyl xanthate. The first one is a high-affinity activation site and the other is a low-affinity inhibition site. Activation of the enzyme in the low concentration of ethyl xanthate arises from increasing the affinity of binding for the substrate as well as increasing the enzyme catalytic constant. The affinity of ligand binding in the activation site is decreased by increasing of the temperature, which is the opposite result for the inhibition site. Hence, the nature of the interaction of ethyl xanthate is different in two distinct sites. The binding process for cresolase inhibition is only entropy driven, meanwhile the binding process for cresolase activation is not only entropy driven but also enthalpy driven means that hydrophobic interaction is more important in the inhibition site.

• Natural Product Sciences
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• 제11권2호
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• pp.89-91
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• 2005

Effect of the neolignans, honokiol (1) and magnolol (2), isolated from Magnoliae Cortex on mushroom tyrosinase activity was investigated in vitro using L-tyrosine as a substrate. Honokiol (1) inhibited tyrosinase activity significantly in a concentration-dependent manner, on the other hand, magnolol (2) did not show tyrosinase inhibitory effect. Honokiol exhibited tyrosinase inhibitory effect with $IC_{50}$ value of $67.9\;{\mu}M$, and proved to act as a non-competitive inhibitor by the analysis of Lineweaver-Burk plot.

• 한방안이비인후피부과학회지
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• 제16권2호
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• pp.96-118
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• 2003

The effect of Glycyrrhizae Radix water extract, known as depigmenting agent, on melanin biosynthesis was investigated in cellular level by using B16 mouse melanoma cells. The inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis was determined by mushroom tyrosinase assay traditionally using in vitro screening test. To determine whether Glycyrrhizae Radix water extract suppress melanin synthesis in cellular level, B16 mouse melanoma cells were cultured in the presence of different concentrations of Glycyrrhizae Radix water extract. Effects on cell proliferation, melanin biosynthesis, tyrosinase activity, DOPAchrome tautomerase activity, and expression level of mRNA for tyrosinase were examined. The maximum concentration of Glycyrrhizae Radix water extract that was not inhibitory to growth of the cells was 2 mgml. At that concentration, melanin synthesis was significantly inhibited without cytotoxicity after 5 days, compared with untreated cells. The treatment with Glycyrrhizae Radix water extract reduced tyrosinase and DOPAchrome tautomerase activity in a dose-dependent manner. However, the treatment with Glycyrrhizae Radix water extract did not affect significantly mRNA levels for tyrosinase. These results suggest that the inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis is correlated with the suppression of tyrosinase and DOPAchrome tautomerase activity more than altering mRNA levels of tyrosinase.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.988-994
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• 2010

The EtOAc fraction of Lespedeza cyrtobotrya showed mushroom tyrosinase inhibitory and radical scavenging activities. Four active compounds were isolated based on Sephadex LH-20 chromatography and HPLC, and the structures were elucidated, on the basis of their LC-MS and NMR spectral data, as 2-(2,4-dihydroxyphenyl)-6-hydroxybenzofuran (1), eriodictyol-7-O-glucopyranoside (2), haginin A (3), and dalbergioidin (4), respectively. Compound (1) showed mushroom tyrosinase inhibitory activity with an $IC_{50}$ value of $5.2\;{\mu}M$ and acted as a competitive inhibitor. Furthermore, $37.3\;{\mu}M$ of compound 1 reduced 50% of the melanin content on human melanoma (MNT-1) cells. The radical scavenging activities of compounds 1, 2, 3, and 4 were shown to have $IC_{50}$ values of 11.0, 24.5, 9.0, and $36.5\;{\mu}M$, respectively, in an ABTS system and $IC_{50}$ values of 42.7, 36.0, 37.7, and $61.7\;{\mu}M$, respectively, in a DPPH system. The mushroom tyrosinase inhibitory activity of the EtOAc fraction of Lespedeza cyrtobotrya was contributed by compounds 1, 3, and 4, and its radical scavenging activity was contributed by compounds 1-4.

• Journal of Microbiology and Biotechnology
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• 제29권8호
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• pp.1204-1211
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• 2019

Fungal exopolysaccharides are important natural products having diverse biological functions. In this study, exopolysaccharides from Fomitopsis castanea mycelia (FEPS) were prepared, and the highest mushroom tyrosinase inhibitory activity was found. FEPS were prepared from cultivation broth by ethanol precipitation method. The extraction yield and protein concentration of FEPS were 213.1 mg/l and 0.03%, respectively. FEPS inhibited mushroom tyrosinase with the half maximal inhibitory concentration ($IC_{50}$) of 16.5 mg/ml and dose-dependently inhibited cellular tyrosinase activity (63.9% at $50{\mu}g/ml$, and 83.3% at $100{\mu}g/ml$) in the cell-free extract of SK-MEL-5 human melanoma cell and ${\alpha}$-melanocyte-stimulating hormone (${\alpha}-MSH$)-stimulated melanin formation in intact SK-MEL-5 human melanoma cell. The $IC_{50}$ of FEPS against NO production from RAW264.7 macrophage cells was $42.8{\pm}0.64{\mu}g/ml$. By in vivo study using a zebrafish model, exposure of FEPS at $400{\mu}g/ml$ to dechorionated zebrafish embryos for 18 h decreased the pigment density, compared to that without FEPS-treated control.

• 대한화장품학회지
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• 제32권3호
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• pp.153-160
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• 2006

본 연구에서는 새로운 미백제를 개발하기 위하여 2',4'-dimethoxyflavone을 합성하였으며 멜라닌 형성 세포에서의 미백효과를 연구하였다 2',4'-Dimethoxyflavone은 프리 라디칼 소거활성(DPPH)이나 mushroom tyrosinase에 대한 직접적인 저해 효과는 없었다. 2',4'-Dimethoxyflavone을 48 h 처리한 B16 멜라노마 세포에서의 멜라닌 생합성 저해효과는 $5{\mu}g/mL$에서는 멜라닌생성이 27% 정도 감소되었다. 2',4'-Dimethoxyflavone은 농도에 비례해서 B16 멜라노마 세포내의 tyrosinase 활성을 저해하였으며, western blot을 이용하여 tyrosinase 단백질 발현도 감소시키는 것으로 확인되었다. 또한, RT-PCR을 이용하여 2',4'-dimethoxyflavone이 멜라닌 생성 과정에 관련하는 유전자 발현을 조사한 결과 tyrosinase와 tyrosinase 관련 단백질(TRP-1)의 mRNA발현을 억제 하였으며 TRP-2의 발현량에는 영향이 없었다. 이 결과로 2',4'-dimethoxyflavone은 멜라닌 형성세포에서 멜라닌 생성에 관여하는 타이로시나제와 TRP-1 유전자 발현 조절을 통하여 미백 효과를 나타내는 것으로 생각된다.

• 대한화장품학회지
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• 제30권1호
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• pp.33-38
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• 2004

본 연구에서는 천마(Gastrodia elata)추출물의 분획과 butanol 분획층의 연속적인 silicagel column chromatography를 통하여 유효성 분인 4-hydroxybenzyl alcohol 1. bis(4-hydroxyphenyl) methane 2. gastrodin(4-$\beta$-D-glucopyranosyloxybenzyl alcohol) 3을 분리하였다. 4-Hydroxybenzyl alcohol 1과 gastrodin(4-$\beta$-D-glucopyranosyloxybenzyl alcohol) 3은 tyrosinase에 대한 저해작용은 없으나 B16 melanoma 세포의 melanin 새성을 억제한다는 것을 발견하였다. Bis(4-hydroxyphenyl)methane 2 ($IC_{50}$/ = 400 $\mu\textrm{g}$/mL)은 arbutin ($IC_{50}$/ = 400 $\mu\textrm{g}$/mL) 보다 약 $\frac{1}{4}$의 tyrosinase 활성 저해작용을 나타내었지만 B16 melanoma 세포의 melanin 생성 억제는 오히려 arbutin 보다 높게 나왔다. 또한 butanol 분획층 ($IC_{50}$/ = 46$\mu\textrm{g}$/mL)의 tyrosinase에 대한 활성 저해 작용이 arbutin ($IC_{50}$/ = 114 $\mu\textrm{g}$/mL)보다 높은 억제 작용을 나타내었고 B16 melanoma 세포의 melanin 생성 억제도 arbutin 보다 높게 나왔다. 특히 butanol 분획층으로부터 분리된 페놀성 혼합물 ($IC_{50}$/ = 2.37 $\mu\textrm{g}$/mL)은 arbutin 보다 약 50배 가까운 매운 높은 tyrosinase 활성 억제 작용을 나타내었다. 이러한 결과로부터 천마추출물로부터 분리된 유효성분들이 tyrosinase에 대한 저해작용 뿐만 아니라 B16 melanoma 세포의 melanin 생성을 억제하는 것을 알 수 있었다.

• 대한화장품학회지
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• 제22권2호
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• pp.193-200
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• 1996

In eastern Asian region, cosmetic preparations containing the molecues possessing the whitening activity are promising. In this study, 100 plant extracts were screened in order to elucidate the whitening effects using in vitro tyrosinase assay and DOPA autooxidation. Several plant extracts such as Morus alba, Glycyrrhiza glabra showed the inhibitory activity (>50%) against mushroom tyrosinase at the concentration 333$mu extrm{m}$/ml. And plant extracts including Morus alba and Bupleurum falcatum showed the inhibitory activity (>50%) at the concentration of 500($\mu\textrm{g}$/ml against DOPA autooxidation. The IC50 values of the several extracts were also found.

• Natural Product Sciences
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• 제29권2호
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• pp.59-66
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• 2023

The anti-melanogenic activity of 259 actinomycete strains was tested, and based on the results for the inhibition of mushroom tyrosinase activity and the reduction in melanin content, Micromonospora sp. JCS1 and JCS7 were selected as the strains with the highest anti-melanogenic potential. The activity-guided fractionation of extracts from JCS1 and JCS7 led to the isolation of the dipeptides cyclo(ʟ-Phenyl alanine (Phe)-ʟ-Proline (Pro)) (1) and cyclo(ʟ-Tryptophan (Trp)-ʟ-Proline (Pro)) (2). These two compounds were tested for their inhibition of mushroom tyrosinase by monitoring ʟ-DOPA levels and melanin production. Cyclo(ʟ-Phe-ʟ-Pro) (1) and cyclo(ʟ-Trp-ʟ-Pro) (2) were thus confirmed to have the potential for use in functional whitening cosmetics containing actinomycete-derived secondary metabolites.