• Title/Summary/Keyword: ${\beta}$-tyrosinase

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Inhibitory Effects of Methanol Extract of Kaempferia galanga on melanogenesis in B16/F10 Melanoma Cells (B16/F10 흑색종양세포에서 삼내자 메탄올 추출물의 멜라닌 생성에 미치는 억제효과)

  • Yoon, Jung-Won;Han, Jung-Min;Yoon, Hwa-Jung;Ko, Woo-Shin
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.1-18
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    • 2013
  • Objective: Recently the demands for the effective and safe depigmentative and anti-aging agents of the skin have increased due to the medical, pharmaceutical and cosmetic reasons. The purpose of this study is to investigate the MKG(Methanol Extract of Kaempferia galanga) and their dermal bioactivity properties related to cosmeceuticals such as depigmentation. Methods: We assessed inhibitory effects of MKG on melanin production in B16/F10 melanoma cells, on mushroom tyrosinase activity, effects of MKG on the expression tyrosinase, TRP-1, TRP-2, GSK-$3{\beta}$, CREB, MITF in B16/F10 melanoma cells without cytotoxicity range. Cell viability was measured by MTT assay and tyrosinase activity was assessed using by DOPA staining, western-blot analysis. We measured inhibition of melanin synthesis and tyrosinase activity by down-regulation of melanogenic enzyme expressions in ${\alpha}$-MSH induced melanogenesis B16/F10 melanoma cells. Results: MKG inhibited tyrosinase-activity, total melanin contents and dendrite out-growth. MKG inhibited melanogenesis by down-regulation of tyorsinase, TRP-1, TRP-2, CREB, and MITF in B16/F10 cells. The treatment with MKG at the 12.5, $25{\mu}g/ml$ level significantly inhibited the melanin synthesis induced ${\alpha}$-MSH in B16/F10 melanoma cells compared with untreated control. Conclusion: These results suggest that MKG inhibit melanin biosynthesis which is involved in hyper-pigmentation. So MKG is considered to be used as a whitening components reducing cytotoxicity.

Production of 3,4-dihydroxyphenyl-L-alanine by Using the ${\beta}$-Tyrosinase of Citrobacter freundii Overexpressed in Recombinant Escherichia coli. (재조합 대장균에서 과발현된 Citorbacter Freundii KCTC2006 유래의 ${\beta}$-Tyrosinase를 이용한 3,4-Dihydroxyphenyl-L-alannine의 생산)

  • Lee, Seung-Goo;Ro, Hyeon-Su;Hong, Seung-Pyo;Lee, Kyu-Jong;Wang, Ji-Won;Tae, Dong-Nyeon;Uhm, Ki-Nam;Bang, Sang-Gu;Kim, Young-Jun;Sung, Moon-Hee
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.44-49
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    • 1996
  • By using the ${\beta}$-tyrosinase of Citrobacter freundii KCT2006, which was cloned and overexpressed in Escherichia coli, 3,4-dihydroxy phenyl-L-alanine (L-DOPA) was synthesized efficiently from pyrocatechol, sodium pyruvate, and ammonium acetate. Optimal temperature and pH for the reaction were determined to be about 18$^{\circ}C$ and 8.5, respectively. The effects of substrate concentrations were also examined at different concentrations of ammonium acetate, sodium pyruvate, and pyrocatechol. Ammoniumacetate and sodium pyruvate increased the reaction rate until the concentrations reached to 300mM and 50mM, respectively. Although pyrocatechol showed the optimal concentration at 20mM, it was controlled between 20mM and 50mM to avoid the depletion of substrate during the enzymatic synthesis. Meanwhile the synthetic rate was improved about 20% when ethanol was included in the reaction solution. Based on above results, a reaction medium for the productin of L-DOPA was prepared and incubated with 1 unit/ml of ${\beta}$-tyrosinase. Pyrocatechol and sodium pyruvate was added to the reaction solutin intermittently to avoid the substrate depletion during the enzymatic reaction. After 24 hour of reaction, 31.6g/l of L-DOPA was accumulated in the reaction solution as soluble and precipitated ones and the conversion yield was about 85.2%.

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Tyrosinase Inhibitory Constituents of Morus bombycis Cortex

  • Kang, Kyo-Bin;Kim, Sang-Du;Kim, Tae-Bum;Jeong, Eun-Ju;Kim, Young-Choong;Sung, Jong-Hyuk;Sung, Sang-Hyun
    • Natural Product Sciences
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    • v.17 no.3
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    • pp.198-201
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    • 2011
  • Tyrosinase is one of the important enzymes in the mammalian melanin synthesis. In the process of melanin synthesis, tyrosine is oxidized to DOPA (3,4-dihydroxyphenylalanine), and DOPA is further oxidized to dopaquinone. Tyrosinase is an enzyme catalyzing this oxidation of tyrosine, so chemicals that inhibit the activity of tyrosinase can be used as skin whitening agents. In this study, we isolated five constituents from the 80% MeOH extract of Morus bombycis cortex by bioactivity-guided fractionation. We performed mushroom tyrosinase inhibition assay. As a result, 7,2',4'-trihydroxyflavanone (1), 2',4',2,4,-tetrahydroxychalcone (2), and oxyresveratrol (3) showed the more potent inhibitory effect compared to kojic acid, a well-known skin whitening agent with antityrosinase effect. Moracinoside M (4) and moracin M-3'-O-${\beta}$-D-glucopyranoside (5) also showed the moderate tyrosinase inhibitory activities.

Synthesis and de-pigmentation effect of phenolic glucoconjugates

  • Kim, Ki-Ho;Kim, Ki-Soo;Lee, Jae-Soeb;Ko, Kang-Il;Lee, Soo-Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.27 no.1
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    • pp.99-109
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    • 2001
  • Novel glucoconjugates phenolic moiety, 3-(methoxycarbonyl)-4-(hydroxyphenyl)-$\beta$-D-glucopyranoside(4), 3-(methoxyacetyl)-4-(hydroxyphenyl)-$\beta$-D-glucopyranoside(7), 4-(hydroxyphenyl)-$\beta$-D-ribofuranoside(11), were synthesized. In order to investigate their depigmentation effect, inhibitory activity against mushroom tyrosinase and inhibitory activity of melanin synthesis in B16 melanoma cell were evaluated in vitro. Compound 11 showed 92.0$\mu\textrm{g}$/㎖ of tyrosinase inhibitory activity whereas compound 4 and 7 showed very low activity not less than 300$\mu\textrm{g}$/㎖. Inhibitory activities of melanin synthesis in B16 melanoma cell of compound 4, 7, and 11 were 8.7, 15.1, and 36.0%, respectively, at the concentration of 100$\mu\textrm{g}$/㎖. Inhibitory activity of compound 11 was much higher than that of arbutin at the same concentration.

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Inhibitory Effect of Soyosangagamhwajae on Melanin Synthesis and its Action Mechanism in B16F10 Mouse Melanoma Cell (소요산가감화제(逍遙散加減化製)의 멜라닌 생성 억제와 작용기전에 관한 연구)

  • Kim, Eun-Seop;Lim, Hyun-Jung;Shin, Sun-Mi;Kim, Soo-Min;Lee, Jung-Eun;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.21 no.1
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    • pp.83-98
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    • 2008
  • Purpose: This study was performed to determine the inhibitory effect of Soyosangagamhwajae(SYG) on melanin synthesis in B16F10 mouse melanoma cell. Methods: The Inhibitory effects of Soyosangagamhwajae(SYG) on melanin synthesis were determined by in-vitro assay. To elucidate inhibitory effects of SYG on melanin synthesis, we determined the melanin release in B16F10 cell. And to investigate the action mechanism, we assessed the gene expression of tyrosinase, TRP-1, TRP-2. PKA, $PKC{\beta}$ in B16F10 cell. Results: 1. SYG significantly inhibited melanin-release in B16F10 cell. 2. SYG significantly inhibited mushroom tyrosinase activity in vitro. 3. SYG significantly suppressed the expression of tyrosinase in B16F10 cell. 4. SYG significantly suppressed the expression of TRP-1, TRP-2 in B16F10 cell. 5. SYG significantly suppressed the expression of PKA, $PKC{\beta}$ in B16F10 cell. Conclusion: From these results, it may be concluded that SYG has the antimelanogenetic effect.

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Determination of tyrosinase inhibitory activity and betanin content changes in beetroot (Beta vulgaris) extracts fermented by EM

  • Yoo, Jong Hee;Kim, Hyun Ki;Yoon, Tae Wou;Mekapogu, Manjulatha;Ahn, Myung Suk;Kwon, Oh Keun;Bang, Keuk Soo;Kim, Yong Ju
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.110-110
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    • 2019
  • Beet (Beta vulgaris) is a crop similar to sugar beet, chard and leaf beets, and its origin is the Mediterranean coast of southern Europe and Central Asia. Among the components contained in beet, betalain, the main component of the root, has been reported to prevent lipid peroxidation induced by active oxygen and free radicals due to its high radical scavenging ability. Among these, the betalain, betanin (Betanidin 5-O-${\beta}$-glucoside) contains both phenolic and cyclic amine groups, all of which are highly electron-donating and act as antioxidants and has tyrosinase inhibitory activity. Betanin accounts for about 75-95% of the total pigment found in the beet. EM stands for effective microorganisms and is a collection of beneficial microorganisms. EM includes yeast, lactic acid bacteria, mycelia, photosynthetic bacteria, actinomycetes, etc. Human patch test according to CTFA guidelines was observed to be a safe source of no stimulation when 5% (v/v) of the EM fermentation liquid was applied to the human body. In addition, beneficial microorganisms are synergistic in the process of co-existence and cultivation and it has the effect of increasing antioxidant capacity and inhibiting corruption. This study confirms the difference in tyrosinase inhibitory activity and betanin content of beetroot extracts and EM fermented beetroot extracts. Hence, these results confirm that EM fermented beetroot extracts are highly beneficial for the human body.

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The Effect of Yukmijihwangtanghapyijihwangagambang on Melanin Synthesis and Related Gene Expressions in B16F10 Mouse Melanoma Cell (육미지황탕합이지환가감방(六味地黃湯合二至丸加減方)이 멜라닌 생성과 관련 유전자 발현에 미치는 영향)

  • Shin, Sun-Mi;Yoo, Dong-Youl
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.4
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    • pp.28-45
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    • 2009
  • Purpose: This study was performed to elucidate the inhibitory effect of Yukmijihwangtanghapyijihwangagambang (YM) on melanin synthesis in B16F10 melanoma cells. Methods: To demonstrate the inhibitory effects of YM on melanin synthesis, we measured the amount of released and produced melanin in B16F10 melanoma call. Also, we evaluated tyrosinase-activity in vitro as well as in B16F10 melanoma call. 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 and MITF in B16F10 melanoma call. Results: 1. YM decreased the release and production of melanin in B16F10 melanoma cells. 2. YM decreased tyrosinase activity in vitro and in B16F10 melanoma cells. 3. YM decreased the expression of tyrosinase, TRP-1, TRP-2 in B16F10 melanoma cells. 4. YM decreased the expression of PKA, $PKC{\beta}$ in B16F10 melanoma cells. 5. YM increased the expression of ERK-1, ERK-2 and AKT-1 in B16F10 melanoma cells. 6. YM decreased the expression of MITF in B16F10 melanoma cells. Conclusion: From these results, it may be concluded that YM has antimelanogenetic effects.

Melanogenesis Inhibitory Activities of Diarylheptanoids from Alnus hirsuta Turcz in B16 Mouse Melanoma Cell

  • Cho, Soo-Min;Kwon, Young-Min;Lee, Jae-Hee;Yon, Kyu-Hyeong;Lee, Min-Won
    • Archives of Pharmacal Research
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    • v.25 no.6
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    • pp.885-888
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    • 2002
  • Four diarylheptanoids, (5R)-1,7-bis (3,4-dihydroxyphenyl)-heptane-5-O-$\beta$-D-glucoside (1), (5R)-1,7-bis (3,4-dihydroxyphenyl)-heptane-5-ol (2), oregonin (3), hirsutanonol (4), were isolated from the bark of Alnus hirsuta Turcz and its inhibitory effects on melanogenesis by measuring the melanin level and tyrosinase activity in B16 melanoma cell were examined. Melanin level and tyrosinase activity were reduced to 75 to 85% by addition of diarylheptanoids to incubation medium of the melanoma cell. On the other hand, melanin level and tyrosinase activity were reduced to 13 to 43% by the addition of diarylheptanoids to incubation medium of the melanoma cell treated with melanogenesis stimulator, $\alpha$-MSH and forskolin. These melanogenesis inhibitory effects were significantly different compared with control.

Purification and Characterization of Neoagarotetraose from Hydrolyzed Agar

  • Jang, Min-Kyung;Lee, Dong-Guen;Kim, Nam-Young;Yu, Ki-Hwan;Jang, Hye-Ji;Lee, Seung-Woo;Jang, Hyo-Jung;Lee, Ye-Ji;Lee, Sang-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1197-1200
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    • 2009
  • The whitening effect, tyrosinase inhibition, and cytotoxicity of neoagarotetraose were measured after its purification from hydrolyzed agar by gel filtration chromatography. In melanoma B16F10 cells, the melanin content of neoagarotetraose-treated cells was the same as that treated by kojic acid or arbutin. In addition, tyrosinase of melanoma cells was strongly inhibited by neoagarotetraose at a concentration of $1{\mu}g/ml$ and similarly inhibited at 10 and $100{\mu}g/ml$ compared with those by arbutin or kojic acid. The activity of mushroom tyrosinase showed a 38% inhibition by neoagarotetraose at $1{\mu}g/ml$, and this inhibitory effect was more efficient than that by kojic acid. Neoagarotetraose revealed a similar $IC_{50}$ (50% inhibition concentration) value for mushroom tyrosinase as that by kojic acid. These data suggest that the neoagarotetraose generated from agar by recombinant $\beta$-agarase might be a good candidate as a cosmetic additive for the whitening effect.

Antioxidant Activities and Tyrosinase Inhibitory Effects of Different Extracts from Pleurotus ostreatus Fruiting Bodies

  • Alam, Nuhu;Yoon, Ki-Nam;Lee, Kyung-Rim;Shin, Pyung-Gyun;Cheong, Jong-Chun;Yoo, Young-Bok;Shim, Mi-Ja;Lee, Min-Woong;Lee, U-Youn;Lee, Tae-Soo
    • Mycobiology
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    • v.38 no.4
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    • pp.295-301
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    • 2010
  • We evaluated the antioxidant activity and tyrosinase inhibitory effects of Pleurotus ostreatus fruiting bodies extracted with acetone, methanol, and hot water. The antioxidant activities were tested against $\beta$-carotene-linoleic acid, reducing power, 1,1-diphenyl-2-picrylhydrazyl free radical scavenging activity, and ferrous chelating ability. Furthermore, phenolic acid and flavonoid contents were also analyzed. The methanol extract showed the strongest $\beta$-carotene-linoleic acid inhibition as compared to the other exracts. The acetone extract (8 mg/mL) showed a significantly high reducing power of 1.54 than the other extracts. The acetone extract was more effective than other extracts for scavenging on 1,1-diphenyl-2-picrylhydrazyl radicals. The strongest chelating effect (85.66%) was obtained from the acetone extract at 1.0 mg/mL. The antioxidant activities of the extracts from the P. ostreatus fruiting bodies increased with increasing concentration. A high performance liquid chromatography analysis detected seven phenolic compounds, including gallic acid, protocatechuic acid, chlorogenic acid, naringenin, hesperetin, formononetin, and biochanin-A in an acetonitrile and 0.1 N hydrochloric acid (5 : 1) solvent extract. The total phenolic compound concentration was $188{\mu}g$/g. Tyrosinase inhibition of the acetone, methanol, and hot water P. ostreatus extracts increased with increasing concentration. The results revealed that the methanol extract had good tyrosinase inhibitory ability, whereas the acetone and hot water extracts showed moderate activity at the concentrations tested. The results suggested that P. ostreatus may have potential as a natural antioxidant.