• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.31-32
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• 2005

Neoagaro-oligosaccharides are produced only by enzymatic degradation of agarose by ${\beta}-agarase.^{1)}$ Neoagaro-oligosaccharides inhibit the growth of bacteria, slow the rate of degradation of starch, are used as low-calorie additives to improve food quality, and have macrophage-stimulating activity. Furthermore, neoagarobiose is a rare reagent that has both moisturizing effect on skin and whitening effect on melanoma $cells.^{2)}$ An agar-degrading marine bacterium was isolated from the sea water at the northeast coast in Cheju island, Korea. The strain was gram negative, aerobic, and motile rod. The 16S rRNA of the strain had the closest match of 98% homology, with that from Agarivorans albus. On the basis of several phenotypic characters and a phylogenetic analysis, this strain was designated Agarivorans sp. JA-1. In solid agar plate, Agarivorans sp. JA-1 produced a diffusible agarase that caused agar softening around the colonies. Agarivorans sp. JA-1 was cultured for 36 hr in marine broth 2216 (Difco, USA) and the supernatant that containing an extracellular ${\beta}-agarase$ was prepared by centrifugation of culture media. The enzyme exhibited relatively strong activity at $40^{\circ}C$ and was stable up to $60^{\circ}C$. Using PCR primers derived from the ${\beta}-agarase$ gene of Vibrio sp., the gene encoding ${\beta}-agarase$ from Agarivorans sp. JA-1 was cloned and sequenced. The structural gene consists of 2931 bp encoding 976 amino acids with a predicted molecular weight of 107,360 Da. The deduced amino acid sequence showed 99% and 34% homology to $agaA^{2)}$ and $agaB^{2)}$ genes for ${\beta}-agarase$ from Vibrio sp., respectively. The expression plasmid for ${\beta}-agarase$ gene of Agarivorans sp. JA-1 is being constructed and the recombinant enzyme will be biochemically characterized.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.9
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• pp.1419-1425
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• 2004

This study was performed to examine in vitro antioxidative activities such as DPPH radical scavenging activity, reducing power and tyrosinase inhibitory effect of various solvent fractions from Chionanthus retusa leaves. Ethyl acetate fraction showed potent antioxidative activity and tyrosinase inhibitory effect. The active compound was isolated from the butanol fraction by silica gel column chromatography and MPLC. The isolated compound was luteolin-4'-O-glucoside determined by $^1H$, $^{13}C$-NMR and 2D NMR. Compared with several antioxidant compounds, luteolin-4'-O-glucoside exhibited effective DPPH radical scavenging activity and reducing power in a concentration dependent manner. Bioassay with pure luteolin-4'-O-glucoside showed a dose-independent inhibitory effect on L-DOPA oxidation by mushroom tyrosinase and its $IC_{50}$ values were established as 23.2 ${\mu}g/mL$. Therefore, we may suggest that luteolin-4'-O-glucoside can be used as a food additive possessing the potent antioxidative activity and skin-whitening cosmetic material.

• Korean Journal of Pharmacognosy
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• v.30 no.4
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• pp.397-403
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• 1999

Tyrosinase plays an important role in the process of melanin polymer biosynthesis. Therefore, the enzyme inhibitors have been of great concern as cosmetics to have skin-whitening effects on the local hyperpigmentation. During the search for new inhibitory compounds on melanin polymer biosynthesis from natural sources, MeOH extracts of 589 higher plants were tested for the inhibitory effect on tyrosinase activity by the muschroom tyrosinase assay in vitro. Among plants tested, the leaves of Cercis chinensis exhibited potent inhibitory effect on mushroom tyrosinase activity. Subsequently seven active compounds were isolated from the ethyl acetate soluble part of acetone extract of the leaves of C. chinensis by the activity guided fractionation monitoring the inhibitory effect on tyrosinase activity. Their chemical structures were identified as $kaempferol-3-0-{\alpha}-L-rhamnoside$, quercitrin, $myricetin-3-0-{\alpha}-L-rhamnoside$, myricetin-3-0-(2'-O-galloyl)- ${\alpha}$ -L-rhamopyranoside (desmanthin), (-)-epicatechin-3-0-gallate, (-)-epigallocatechin-3-0-gallate, and methyl gallate on the basis of the speculation of spectral data and chemical reaction. Among the flavonol rhamnosides, myricetin-3-0-(2'-O-galloyl)- -L-rhamnoside(desmanthin) showed most potent inhibitory effect on tyrosinase activity and the structure of B-ring in flavonol moiety was related to the activity. (-)-Epigallocatechin-3-O-gallate having pyrogallol group in flavan-3-ol moiety exhibited more potent inhibitory effect than (-)-epicatechin-3-0-gallate having catechol group in flavan-3-ol moiety on mushroom tyrosinase activity.

• Journal of the Korean Applied Science and Technology
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• v.34 no.3
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• pp.602-612
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• 2017

This study was performed to verify various effects of 70% ethanol extract and distilled water extract of pine leaf such as antioxidant, anti-inflammatory, skin-lightening effects analysis in cells. The result of antioxidant experiment showed that polyphenol content increased concentration dependently in polyphenol and flavonoid effects. The total content of polyphenols and flavonoids was much higher in ethanol extract than water extract. In all the cells of B16F10, RAW264.7 cytotoxicity at each concentration level didn't appear. The result of measuring NO production inhibition showed that it inhibited NO production led to LPS effectively, so anti-inflammatory activity was confirmed. The result of measuring melanin biosynthesis inhibition showed that there was considerably reduction effect, but it performed western blot, and as a result of MITF, Tyrosinase protein revelation, the inhibition of MITF, Tyrosinase revelation was confirmed concentration dependently, Therefore, in this study pine leaf extract was expected to be used as a cosmetic ingredient.

• Journal of Microbiology and Biotechnology
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• v.25 no.12
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• pp.2011-2015
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• 2015

Ginsenoside Rb2 (Gin-Rb2) was purified from the fruit extract of Panax ginseng. Its chemical structure was measured by spectroscopic analysis, including HR-FAB-MS, ¹H-NMR, and IR spectroscopy. Gin-Rb2 decreased potent melanogenesis in melan-a cells, with 23.4% at 80 μM without cytotoxicity. Gin-Rb2 also decreased tyrosinase and MITF protein expression in melan-a cells. Furthermore, Gin-Rb2 presented inhibition of the body pigmentation in the zebrafish in vivo system and reduced melanin contents and tyrosinase activity. These results show that Gin-Rb2 isolated from P. ginseng may be an effective skin-whitening agent via the in vitro and in vivo systems.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.2
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• pp.215-219
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• 2014

In Korean Cosmetic Act, four uses of functional cosmetics are recognized: skin whitening, anti-wrinkle, UV protection or suntan. Cosmeceuticals, one of the functional cosmetics, have been specifically developed for medical and cosmetic benefits. However, the uses of cosmeceuticals in patent applications are not limited to those of functional cosmetics, and the number is increasing with variety of medical use claims such as anti-inflammatory, prevention of hair loss, or obesity in cosmeceutical patent applications. Since some of the cosmeceutical use claims are not clearly distinguished from medical use claims in pharmaceuticals, there has been a controversy over the extent of granting patents regarding the medical use of cosmeceuticals. In this article, we have investigated the range of medical uses in patented claims of functional cosmetics, summarized examiners' views and discussed the examination practice of cosmetic-related inventions in the United States, Europe, and Japan.

• Biomolecules & Therapeutics
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• v.26 no.3
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• pp.282-289
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• 2018

Melanin is a pigment produced from tyrosine in melanocytes. Although melanin has a protective role against UVB radiation-induced damage, it is also associated with the development of melanoma and darker skin tone. Tyrosinase is a key enzyme in melanin synthesis, which regulates the rate-limiting step during conversion of tyrosine into DOPA and dopaquinone. To develop effective RNA interference therapeutics, we designed a melanin siRNA pool by applying multiple prediction programs to reduce human tyrosinase levels. First, 272 siRNAs passed the target accessibility evaluation using the RNAxs program. Then we selected 34 siRNA sequences with ${\Delta}G{\geq}-34.6kcal/mol$, i-Score value ${\geq}65$, and siRNA scales score ${\leq}30$. siRNAs were designed as 19-bp RNA duplexes with an asymmetric 3' overhang at the 3' end of the antisense strand. We tested if these siRNAs effectively reduced tyrosinase gene expression using qRT-PCR and found that 17 siRNA sequences were more effective than commercially available siRNA. Three siRNAs further tested showed an effective visual color change in MNT-1 human cells without cytotoxic effects, indicating these sequences are anti-melanogenic. Our study revealed that human tyrosinase siRNAs could be efficiently designed using multiple prediction algorithms.

• 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.

• Journal of the Korean Applied Science and Technology
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• v.26 no.3
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• pp.269-278
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• 2009

In this study, the anti oxidative effects, inhibitory effects on elastase, and components of Securinega suffruticosa extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) and reactive oxygen species ROS) scavenging activities ($OSC_{50}$) of extract/fractions from Securinega suffruticosa were measured. The aglycone fraction ($9.04\;{\pm}\;0.51\;{\mu}g/mL$) and 50 % ethanol extract ($1.05\;{\pm}\;0.41\;{\mu}g/mL$) showed the most effective scavenging activities. The protective effects on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The aglycone fraction exhibited the most prominent cellular protective effect (${\tau}_{50},\;102.77\;{\pm}\;5.37$ min at $10{\mu}g/mL$). The inhibitory effect of ethyl acetate fraction on tyrosinase were of examined ($210.0\;{\pm}\;1.02\;{\mu}g/mL$). The inhibitory effect of aglycone fraction on elastase were also investigated ($17.6\;{\pm}\;1.26\;{\mu}g/mL$). These results indicate that extract/fractions of Securinega suffruticosa can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS. Securinega suffruticosa extract could be used as a new cosmeceutical ingredient for whitening and anti-wrinkle products.

• Nutrition Research and Practice
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• v.12 no.1
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• pp.3-12
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• 2018

BACKGROUND/OBJECTIVES: Sageretia thea is traditionally used as a medicinal herb to treat various diseases, including skin disorders, in China and Korea. This study evaluated the inhibitory effect of Sageretia thea fruit on melanogenesis and its underlying mechanisms in B16F10 mouse melanoma cells. The active chemical compounds in anti-melanogenesis were determined in Sageretia thea. MATERIALS/METHODS: Solvent fractions from the crude extract were investigated for anti-melanogenic activities. These activities and the mechanism of anti-melanogenesis in B16F10 cells were examined by determining melanin content and tyrosinase activity, and by performing western blotting. RESULTS: The n-hexane fraction of Sageretia thea fruit (HFSF) exhibited significant anti-melanogenic activity among the various solvent fractions without reducing viability of B16F10 cells. The HFSF suppressed the expression of tyrosinase and tyrosinase-related protein 1 (TRP1). The reduction of microphthalmia-associated transcription factor (MITF) expression by the HFSF was mediated by the Akt/glycogen synthase kinase 3 beta ($GSK3{\beta}$) signaling pathway, which promotes the reduction of ${\beta}-catenin$. Treatment with the $GSK3{\beta}$ inhibitor 6-bromoindirubin-3'-oxime (BIO) restored HFSF-induced inhibition of MITF expression. The HFSF bioactive constituents responsible for anti-melanogenic activity were identified by bioassay-guided fractionation and gas chromatography-mass spectrometry analysis as methyl linoleate and methyl linolenate. CONCLUSIONS: These results indicate that HFSF and its constituents, methyl linoleate and methyl linolenate, could be used as whitening agents in cosmetics and have potential for treating hyperpigmentation disorders in the clinic.