• Applied Biological Chemistry
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• 제22권4호
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• pp.191-197
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• 1979

Km값 및 분광학적성질(分光學的性質)를 조사하고 아울러 이 효소(酵素)의 결정화법(結晶化法)을 검토하여 다음과 같은 결과를 얻었다. 1. 효소(酵素)의 정제(精製)과정에서 유안분별침전(硫安分別沈澱)을 한 조효소(粗酵素)를 pH6.0 및 7.0의 인산염(燐酸鹽)완충액으로 48시간 이상의 충분한 투석(透析)을 시키고 DEAE-Sephadex column chromatography에서는 11mg protein/ml DEAE-Sephadex 정도의 DEAE-Sephadex를 사용하는 것이 효과적이였다. 2. 효소(酵素)의 정제과정(精製過程)에서 이미 알려진 protamin 황산(黃酸) 처리와 Sephadex G-150의 gel여과는 생략하여도 무방하였다. 3. 효소(酵素)의 결정화(結晶化)에서는 단백질을 20mg/ml의 농도가 되게끔 2-mercaptoethanol를 함유하는 0.01M 인산(燐酸)카리 완충액에 녹여 고체유안(固體硫安) 첨가(添加)법으로 결정(結晶)시키는 것이 가장 큰 육각봉상(六角棒狀)의 결정(結晶)이 얻어졌다. 4. Pyridoxal phosphate와 결합(結合)한 holo형 ${\beta}-tyrosinase$는 340nm와 430nm의 파장(波長)에서 각각 최대흡수(最大吸收)를 나타내었다. 5. ${\beta}-tyrosinase$의 L-tyrosine에 대한 Km값은 $2.31{\times}10^{-4}M$이였으며 SDS-polyacrylamide 전기영동법(電氣泳動法)에 의한 이 효소(酵素)의 subunit의 분자량(分子量)은 약 5,000이였다.

• 생약학회지
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• 제32권1호통권124호
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• pp.68-71
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• 2001

For the use of tannins in the whitening-effect cosmetics, inhibitory effect against tyrosinase activity was determined. Three condensed tannins including gallocatechin, gallocatechin 3',4'-di-O-gallate and epicatechin 3-O-gallate and three hydrolyzable tannins, 1,2,6-tri-O-galloyl-${\beta}$-D-glucose, 2,3-(S)-HHDP-D-glucose and pedunculagin showed 15-29% mild inhibitory effects against tyrosinase activity.

• Preventive Nutrition and Food Science
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• 제12권2호
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• pp.119-121
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• 2007

The tyrosinase inhibition activity and mutagenicity as assessed by the Ames test on phenolic antioxidants (5-Caffeoyl quinic acid, 3,4-Dihydroxy cinnamic acid, Quercetin 3-O-${\beta}$-D-glucopyranose, Kaempferol 3-O-${\beta}$- D-glucopyranose) and the ethyl acetate fraction isolated from mulberry leaves were examined. The ethyl acetate fraction and chlorogenic acid exhibited weaker tyrosinase inhibitory activities than kojic acid. In addition, the ethyl acetate fraction from mulberry leaves, containing phenolic antioxidants, showed no mutagenicity by the Ames test.

• Natural Product Sciences
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• 제22권2호
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• pp.111-116
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• 2016

Phytochemical investigation of the stems of Pueraria lobata (Wild) Ohwi (Leguminosae), led to the isolation of eighteen known compounds: ${\beta}$-amyrone (1), (+)-pinoresinol (2), (+)-syringaresinol (3) $(+)-syringaresinol-O-{\beta}-{\small{D}}-glucoside$ (4), (+)-lariciresinol (5), (-)-tuberosin (6), naringenin (7), liquiritigenin (8), isoliquiritigenin (9) genistein (10), daidzein (11) daidzin (12) daidzein 4',7-diglucoside (13) 2,4,4'-trihydroxy deoxybenzoin (14), S-(+)-1-hydroxy-3-(4-hydroxyphenyl)-1-(4-hydroxy-2-methoxy-phenyl)propan-2-one (15), methyl $2-O-{\beta}-{\small{D}}-glucopyranosylbenzoate$ (16), pyromeconic acid $3-O-{\beta}-{\small{D}}-glucopyranoside$ 6'- (O-4''-hydroxy-3-methoxybenzoate) (17), and allantion (18). The chemical structures of these compounds were elucidated from spectroscopic data and by comparison of those data with previously published results. The effects of isolated compounds on mushroom tyrosinase enzymatic activity were screened. The results indicated that, chloroform extract of P. lobata stems turned out to be having tyrosinase inhibitory effect, and only compounds 5, 8, 9, and 11 showed enzyme inhibitory activity, with $IC_{50}$ values of $21.49{\pm}4.44$, $25.24{\pm}6.79$, $4.85{\pm}2.29$, and $17.50{\pm}1.29{\mu}M$, respectively, in comparison with these of positive control, kojic acid ($IC_{50}\;12.28{\pm}2.72{\mu}M$). The results suggest that P. lobata stems extract as well as its chemical components may represent as potential candidates for tyrosinase inhibitors.

• 대한화장품학회지
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• 제35권3호
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• pp.243-250
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• 2009

천연 미백 소재 개발을 위하여 복령피 추출물과 복령피 추출물에서 분리한 활성 물질의 멜라닌 생성에 연관된 생리 활성을 분석하였다. 복령피 추출물은 $100{\mu}g$/mL 이하에서 세포 독성이 없는 것으로 확인 되었으며 free radical 소거능(DPPH)과 superoxide radicals 소거능 결과는 각각 $IC_{50}=19.4{\pm}2.21{\mu}g$/mL, $IC_{50}=103{\pm}3.33{\mu}g$/mL을 나타내었다. $50{\mu}g$/mL 농도에서 B16 melanoma 세포 내 tyrosinase의 활성을 34 % 저해 하였으며, 세포 내 멜라닌 생합성 저해 효과는 $50{\mu}g$/mL 농도의 복령피 추출물을 72 h 동안 처리한 세포에서 51 % 억제율을 보이는 것으로 나타났다. 이러한 복령피 추출물의 활성 물질을 분리하여 $^1H$-NMR, $^{13}C$-NMR, Mass analysis 등의 기기 분석을 실시한 결과 triterpene류의 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid로 동정되었고 $100{\mu}g$/mL 이하에서 세포 독성이 없는 것으로 확인 되었다. 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid의 free radical 소거능과 superoxide radicals 소거능 결과는 각각 $IC_{50}=4.3{\pm}0.15{\mu}g$/mL, $IC_{50}=54{\pm}1.67{\mu}g$/mL을 나타내었다. $10{\mu}g$/mL 농도에서 세포내 tyrosinase의 활성을 43 % 저해 하였으며, 멜라닌 저해 효과를 확인한 결과 $IC_{50}=3.6{\mu}g$/mL으로 나타났다. Western blot을 이용하여 tyrosinase, tyrosinase related protein-1 (TRP-1), TRP-2 단백질의 발현 감소를 확인하였다. 복령피 추출물과 3-$\beta$-hydroxylanosta-7,9(11),24-trien-4-oic acid는 우수한 미백 효능을 갖는 화장품 소재로의 개발 가능성이 클 것으로 기대된다.

• Preventive Nutrition and Food Science
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• 제2권4호
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• pp.285-290
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• 1997

Previously, a 50% aqueous methanol extract of Galla rhois was shown to be the most potent tyrosinase inhibition activity with an {TEX}$IC_{50}${/TEX}(the concentration causing 50% inhibition of tyrosinase activity) of 0.2mg/ml of 205 crude drug extracts. To isolate tyrosinase inhibitors, the methanol extract was evaporated to a small volume in vacuo, and then partitioned stepwise with benzene and ethyl acetate(EtOAc). the EtOAc fraction was solubilized in 10% MeOH solution, and then fractionated successively by Diaion HP-20 and Sephadex LH-20 column chromatography, and preparative HPLC. Three phenolic compounds were isolated, and characterized as gallic acid(GA), methyl gallate(MG) and 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose(PGG) by UV, IR, {TEX}${1}^H${/TEX}-&{TEX}${13}^C${/TEX}-NMR, and FAB-MS spectroscopy, PGG({TEX}$IC_{50}${/TEX}=50$\mu\textrm{g}$/ml) showed a considerable inhibitory effect against mushroom tyrosinase, while GA({TEX}$IC_{50}${/TEX}=1.6mg/ml) and MG({TEX}$IC_{50}${/TEX}=234$\mu\textrm{g}$/ml) did not show an appreciable effect. Meanwhile, MG inhibited greatly melanogenesis in a murine melanocyte cell line, Mel-Ab. MG and PGG showed typical noncompetitive inhibition patterns against mushroom tyrosinase. These results suggest that PGG and MG may be potentially useful as either anti-browning or anti-melanogenic agents in foods and cosmetics.

• Natural Product Sciences
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• 제8권4호
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• pp.183-185
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• 2002

To investigate skin whitening natural substances, the effects on melanogenesis by measuring the tyrosinase activity and the melanin contents of three hydrolysable tannins, $1,2,6-tri-O-galloyl-{\beta}-D-glucose$ (1), 2,3-(S)-HHDP-D-glucose (2) and pedunculagin (3) in B16 melanoma cells were examined. $1,2,6-Tri-O-galloyl-{\beta}-D-glucose$ (1), 2,3-(S)-HHDP-D-glucose (2) and pedunculagin (3) inhibited tyrosinase activity in B16 melanoma cells in a dose-dependent manner.

• Preventive Nutrition and Food Science
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• 제7권4호
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• pp.447-450
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• 2002

Previously, a methanol extract from seeds of Paeonia lactiflora was shown to have a potent inhibitory activities against tyrosinase and soybean lipoxygenase (SLO). Seven stilbenes, trans-resveratrol-4-Ο-$\beta$-D-glucoside, trans resveratrol, trans-$\varepsilon$-viniferin, cis-$\varepsilon$-viniferin, gnetin H, suffruticosol A and B were isolated from the seeds as active principles for inhibition of the enzymatic activity. Among them, the resveratrol trimer, gnetin H exhibited the most potent inhibitory activities against tyrosinase and SLO, respectively. Additionally, the resveratrol dimers, trans-$\varepsilon$-viniferin and cis-$\varepsilon$-viniferin exhibited significant inhibitory activity against the two oxidative enzymes. Meanwhile, three other stilbene derivatives, such as trans-resveratrol, suffruticosol A and suffruticosol B had also weak inhibition activity. The least inhibitory activity was observed in transresveratrol-4-Ο-$\beta$-D-glucoside. These results suggest that resveratrol dimers and trimer in the seeds of Paeonia lactiflora are potentially useful therapeutic agents against pathological disorders such as hyperpigmentation and inflammation.

• 대한한방부인과학회지
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• 제22권2호
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• pp.43-59
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• 2009

Purpose: This study was performed to determine the inhibitory effect of Persimmon Leaves extract (PL) on melanin synthesis in B16F10 melanoma cells B16F10. Methods: The inhibitory effects of PL on melanin synthesis were determined by in vitro assay. To elucidate inhibitory effects of PL on melanin synthesis, we determined the melanin release and melanin production in B16F10. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, PKA, PKC${\beta}$, ERK-1, ERK-2, AKT-1, MITF in B16F10. Results: 1. PL inhibited melanin release, melanin production in B16F10. 2. PL inhibited tyrosinase activity in vitro and in B16F10. 3. PL suppressed the expression of tyrosinase, TRP-1, TRP-2 in B16F10. 4. PL suppressed the expression of PKA, PKC${\beta}$ in B16F10. 5. PL increased the expression of ERK-1, ERK-2, AKT-1 in B16F10. 6. PL suppressed the expression of MITF in B16F10. Conclusion: From these results, it may be concluded that PL is possesed of the antimelanogenetic effects.

• 대한한방부인과학회지
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• 제22권4호
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• pp.1-18
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• 2009

Purpose: This study was performed to elucidate the inhibitory effect of Mibaeksan (MB) on melanin synthesis in B16F10 mouse melanoma cell. Methods: To demonstrate the inhibitory effects of MB on melanin synthesis, we measured the amount of released and produced melanin in B16F10 melanoma cell. Also, we evaluated tyrosinase-activity in vitro as well as in B16F10 melanoma cell. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2, MMP-2, PKA, $PKC{\beta}$, ERK-1 ERK-2, AKT-1 and MITF in B16F10 melanoma cells. Results: 1. MB decreased the release and production of melanin in B16F10 melanoma cells. 2. MB decreased tyrosinase activity in vitro and in B16F10 melanoma cells. 3. MB decreased the expression of tyrosinase, TRP-1, TRP-2, PKA, $PKC{\beta}$ and MMP-2 in B16F10 melanoma cells. 4. MB increased the expression of ERK-1, ERK-2 and AKT-1 in B16F10 melanoma cells. 5. MB decreased the expression of MITF in B16F10 melanoma cells. Conclusion: From these results, it may be concluded that MB has the antimelanogenetic effects.