• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.2
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• pp.87-92
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• 2007

To develop the skin whitening agent, we investigated the effects of Acanthopeltis japonica, a rhodophyta on the coast of Jeju island, on melanogenesis. Dried A. japonica was refluxed with 70 % aqueous ethanol and the extract was evaporated to dryness. To validate the activity as a depigmenting agent, various in vitro tests, polyphenol contents, and free radical scavenging activity were performed. In addition, cellular tyrosinase activity and protein expression of p-ERT, tyrosinase, TRP-1, and TRP-2 were measured in B16/F10 murine melanoma cells. A. japonica had low polyphenol contents and low free radicals scavenging activities against 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical. A. japonica suppressed cellular tyrosinase activity up to 86.9 % at $100{\mu}g/mL$ with inhibition or tyrosinase and TRP-1 expression in ${\alpha}$-melanocyte stimulating hormone (${\alpha}$-MSH)-treated B16/F10 melanoma cells. Our results suggest that inhibitory effects of A. japonica on melanogenesis are due to inhibiting the pathways involving ${\alpha}$-MSH-induced ERK activation. Therefore, A. japonica nay be useful as a skin whitening agent associated with the suppressive effect of melanotrophin-induced signaling pathway to inhibit melanin synthesis.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1665-1672
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• 2012

An 80% ethanol extract of Hizikia fusiforme was obtained and followed by successive fractionation using the organic solvents n-hexane, ethyl acetate, and n-butanol to identify the antioxidative substance. The aqueous part of the nbutanol fractionation step, showing high antioxidative activity, was subjected to reverse-phase liquid chromatography. As a result, a substance purified from a BB-2 fraction showed high antioxidative activity. The m/z 419 [M+H] molecular ion peak in the fraction was observed by the analysis of the ESI-LC/MS spectrum. By the analysis of 1H NMR (500 MHz, DMSO-$d_6$) and $^{13}C$ NMR (125 MHz, DMSO-$d_6$) spectra, a unique compound of the fraction was biochemically identified as a 5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone (5HHMF). We also investigated the effect of 5HHMF on human gastric AGS carcinoma cells. Western blot analysis suggested that the flavone substantially increased the levels of the death receptor-associated apoptosis mediators Fas, Fas L, FADD, TRADD, and DR4 in a concentration-dependent manner. The levels of Fas, Fas L, TRADD, and DR4 in the cells treated with 5HHMF ($5{\mu}g/ml$) were approximately 26.4-, 12.8-, 6.7-, and 9.8-times higher than those of non-treated cells, respectively. Of note, the level of FADD protein in the cells exposed to 5HHMF ($1{\mu}g/ml$) increased approximately 9.6-times. In addition, the cleavage of caspase-3, -8, and -9 in cultured AGS cells treated with 5HHMF was significantly confirmed. Therefore, our results suggest that 5HHMF from H. fusiforme is involved in the induction of death receptor-associated apoptosis mediators in human gastric AGS carcinoma cells.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.305-310
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• 2005

To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.41 no.12
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• pp.1740-1746
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• 2012

Sparassis (S.) crispa is an edible mushroom abundant in dietary fiber and ${\beta}$-glucan. The aim of this study was to prepare extracts and residues of the fruit bodies of S. crispa fermented with Lactobacillus (L.) brevis and Monascus (M.) pilosus and to measure the remaining dietary fiber and ${\beta}$-glucan. Dried powder of S. crispa containing 64.4 g/100 g total dietary fiber (2.6 g/100 g soluble and 61.8 g/100 g insoluble dietary fibers) and 24.0 g/100 g ${\beta}$-glucan was used as the starting material for the extraction. Raw and fermented S. crispa were extracted with hot water and three kinds of aqueous ethanol (50, 70, and 90%, v/v), respectively. A hot water extract from S. crispa fermented with M. pilosus had greater soluble dietary fiber content (19.3 g/100 g) than that from raw S. crispa with 14.6 g/100 g soluble dietary fiber or that from L. brevis-fermented S. crispa with 8.2 g/100 g soluble dietary fiber. The yield of the extract was 16.6% of intial weight of dried S. crispa. After hot water extraction of S. crispa fermented with M. pilosus, residues containing 90.5 g/100 g total dietary fiber (1.3 g/100 g soluble and 89.2 g/100 g insoluble dietary fibers) were obtained, and the yield was 69.6% of intial weight of dried S. crispa. The residue (31.0 g/100 g) contained more ${\beta}$-glucan than raw S. crispa or M. pilosus-fermented S. crispa (24.4 g/100 g). The resulting hot water extract and residue from S. crispa fermented with M. pilosus would be suitable for use in preparing liquid and powdered health functional foods, respectively.

• Food Science and Preservation
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• v.16 no.2
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• pp.259-265
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• 2009

To develop an effective anti-hangover product, hot-water extracts of 25 medicinal herbs were screened for inhibition or activation of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH), and 12 herbs were selected for further study. Chosen medicinal herb extracts(CMHEs) were fermented by Lactobacillus delbruechii subspecies lactis for 10 days at $35^{\circ}C$ after saccharification with nuruk(malt inoculated by 5 types of microbs) for 72 hours at $35^{\circ}C$ and both CMHEs and fermented CMHEs(FCMHEs) were explored for anti-hangover effects in vitro. We found significant ADH inhibition by hot-water extracts of Pueraria thunbergiana, Hovenia dulcis Thunb, Lycium chinense, Glycyrrhiza uralensis, Acanthopanax sessiliflorus, Liriope platyphylla, and Ixeris dentata, and significant ALDH activation by extracts of Acanthopanax sessiliflorus, Lycium chinense, Ixeris dentata, and Polypori umbellati of the Polyporaceae. The ADH effects on CMHE and FCMHE were -20.22% and -62.63% of control values, and the ALDH effects 173.20% and 280.17%, respectively. In rats given 20%(v/v) alcohol(15 mL/kg), FCMHEs significantly decreased blood acetaldehyde concentrations on 3 hours after ethanol administration, in a dose-dependent manner(p<0.05). Notably, blood acetaldehyde concentrations were markedly reduced in animals given FCMHEs(400 mg/kg) compared to levels seen in rats receiving CADB(commercial alcohol detoxification beverage). Thus, anti-hangover effects were promoted by fermentation of certain medicinal herb extracts.

• Korean Journal of Medicinal Crop Science
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• v.13 no.4
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• pp.199-205
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• 2005

This study was carried out to investigate the effect of 70% ethanol extract and each fraction from Rhododendron brachycarpum D. Don leaves on cytotoxicity, anticancer, genotoxicity and immunological activity in vitro bioassay. Cytotoxicity for human normal cells (HEL299 and Chang) of the samples was shown below 35% in 0.5 mg/ml concentration of samples except aqueous fraction by SRB assay. DNA damage on the Chang cell of the samples alone in comet assay was observed very weak damage activity even in high concentration (1 mg/ml) of the samples. The anticancer effect of the samples on human cancer cell lines (A549, AGS, Hep3B, MCF7) was indicated that the cancer cells were inhibited gradually in proportion to the increase of the concentration of the samples by MTT assay. The growth of the Raji and Jurkat cells were hastened by adding butanol fraction among the samples. In the genotoxicity on $H_2O_2-induced$ DNA damage in Chang cells using alkaline comet assay, most of samples were shown a strong protective activity from DNA OTM values.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.118-124
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• 2009

Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.