• Microbiology and Biotechnology Letters
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• v.37 no.1
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• pp.55-61
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• 2009

Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having $\beta$-glucosidase (to transform ginsenoside $Rb_1$) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-$886^T$. The strain, Gsoil 235, could transform ginsenoside $Rb_1$ into Rd, $Rg_3$ and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for, $\beta$-glucosidase was nutrient broth (In 48 hours, almost ginsenoside $Rb_1$ could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.3
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• pp.139-144
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• 2007

In the previous study, we reported the antioxidative activity of Equisetum arvense extracts. In this study, its inhibitory effect on elastase and components were investigated. Aglycone fractions obtained from the deglycosylation reaction of ethylacetate fraction among the Equisetum arvense extracts, showed 4 bands and 4 peaks in TLC and HPLC experiments, respectively. Four components were identified as luteolin(composition ratio, 19.12%), quercetin(12.87), apigenin(15.81) and kaempferol(52.20). TLC chromatogram of ethylacetate fraction of Equisetum arvense extract revealed 7 bands and HPLC chromatogram showed 8 peaks, which were identified as kaempferol-3,7-O-diglucoside(composition ratio, 15.74%), luteolin-5-O-glucoside(galuteolin, 11.91), apigenin-5-O-glucoside(12.91), kaempferol-3-O-glucoside(astragalin, 27.94), quercetin-glycoside(10.81, structure was not determined), kaempferol-glycoside (12.33, structure was not determined), luteolin(3.72) and apigenin(4.62) in the order of elution time. The inhibitory effect of aglycone fraction on elastase($IC_{50}$, $9.8{\mu}g/mL$) was very high. But ethylacetate fraction(flavonoid glycosides) rarely exhibited the inhibitory activity on elastase. Combined with the previous results of the antioxidative activity of Equisetum arvense extracts, it is concluded that the inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

• Korean Journal of Food Science and Technology
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• v.40 no.3
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• pp.343-348
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• 2008

The ovomucoid (OM) of egg whites is recognized as a major allergen. Here, the allergenicity of OM in egg whites (EW) treated by chemical, enzymatic, and physiological methods were investigated by competitive inhibitory ELISA using human IgE antibody acquired from egg-allergic patients. Enzymatic hydrolysis, irradiation, and succinic anhydride treatments did not reduce the allergenicity of the OM effectively. Allergenicity was reduced to only 1/20 by deglycosylation with trifluoromethanesulfonic acid (TFMS). Heat treatment of the OM at $121^{\circ}C$ for 10 min reduced allergenicity to 1/100. Furthermore, NaOH (over 3%) treatment reduced allergenicity to 1/10,000, and the combinatory treatment of NaOH (over 0.3%) and heat ($70^{\circ}C$, 15 min) reduced it to less than 1/10,000, which was the most effective method. In this study, which analyzed treated EW using ELISA and patient-derived IgE, the OM allergenicity was nearly the same as its antigenicity according to ELISA using rabbit IgG. However, in the case of the TFMS-treated EW, the antigenicity was much lower than the allergenicity. These results suggest that the allergenicity of OM is slightly different from its antigenicity.

• Journal of Ginseng Research
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• v.42 no.4
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• pp.504-511
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• 2018

Background: The biological activities of ginseng saponins (ginsenosides) are associated with type, number, and position of sugar moieties linked to aglycone skeletons. Deglycosylated minor ginsenosides are known to be more biologically active than major ginsenosides. Accordingly, the deglycosylation of major ginsenosides can provide the multibioactive effects of ginsenosides. The purpose of this study was to transform ginsenoside Rb2, one of the protopanaxadiol-type major ginsenosides, into minor ginsenosides using ${\beta}$-glycosidase (BG-1) purified from Armillaria mellea mycelium. Methods: Ginsenoside Rb2 was hydrolyzed by using BG-1; the hydrolytic properties of Rb2 by BG-1 were also characterized. In addition, the influence of reaction conditions such as reaction time, pH, and temperature, and transformation pathways of Rb2, Rd, F2, compound O (C-O), and C-Y by treatment with BG-1 were investigated. Results: BG-1 first hydrolyzes 3-O-outer ${\beta}$-$\text\tiny{D}$-glucoside of Rb2, then 3-O-${\beta}$-$\text\tiny{D}$-glucoside of C-O into C-Y. C-Y was gradually converted into C-K with a prolonged reaction time, but the pathway of Rb2 ${\rightarrow}$ Rd ${\rightarrow}$ F2 ${\rightarrow}$ C-K was not observed. The optimum reaction conditions for C-Y and C-K formation from Rb2 by BG-1 were pH 4.0-4.5, temperature $45-60^{\circ}C$, and reaction time 72-96 h. Conclusion: ${\beta}$-Glycosidase purified from A. mellea mycelium can be efficiently used to transform Rb2 into C-Y and C-K. To our best knowledge, this is the first result of transformation from Rb2 into C-Y and C-K by basidiomycete mushroom enzyme.

• Journal of Life Science
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• v.20 no.5
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• pp.683-690
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• 2010

The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.49 no.3
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• pp.259-267
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• 2023

This study was conducted to evaluate physiological activity of flower extract of peony (Paeonia lactiflora Pall.) by hydrolysis and to use it as a valuable cosmetic ingredients. Four cultivar of peony flowers were extracted, and the highest active ingredient contents was selected, and that cultivar was used for hydrolyzing. The results showed that high concentration of hydrochloric acid (HCl) hydrolyzed, and biological hydrolysis using enzymes had no activity. The deglycosylation of peonidin 3,5-diglucoside occurred by hydrolysis. The hydrolysate contains 63.3 ppm of peonidin, a red-colored anthocyanin compound. The antioxidant activity of hydrolysate was compared with extract. The results showed the strong antioxidant activity in hydrolysate (96%) than extract (82%). In addition, hydrolysate of peony flower showed higher inhibitory activity of NO release than extract. UVA assay using fibroblast cell (CCD-986Sk) showed that hydrolysate increased cell viability than extract under UVA exposure. Based on these results, we anticipate that hydrolysate of peony flower can be used a valuable cosmetic ingredient.

• Microbiology and Biotechnology Letters
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• v.37 no.2
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• pp.99-104
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• 2009

The $\beta$-glucosidase gene from Streptomyces coelicolor A3(2) was cloned and expressed in Escherichia coli. The ORF consisted of 1377 nucleotides encoding 51 kDa in a predicted molecular weight. Effects of pH indicated that the $\beta$-glucosidase showed similar activity using $\alpha$-pNPG($\rho$-nitrophenyl-$\alpha$-D-glucopyranoside), $\beta$-pNPG($\rho$-nitrophenyl-$\beta$-D-glucopyranoside), and $\beta$-pNPF($\rho$-nitrophenyl-$\beta$-D-fucopyranoside) at range of pH 3 to 10, and high activity using $\beta$-pNPGA ($\rho$-nitrophenyl-$\beta$-D-galactopyranoside) from pH 5 to 10, especially, 3.3 times higher activity at pH 9. Effects of temperature indicated that the $\beta$-glucosidase showed low activity using $\alpha$-pNPG, $\beta$-pNPG, and $\beta$-pNPF from $20^{\circ}C$ to $70^{\circ}C$, and increased activity using $\beta$-pNPGA from $30^{\circ}C$ to $50^{\circ}C$, 1.8 times higher activity at $50^{\circ}C$ than at $30^{\circ}C$. According to activity determination of other substrates, the enzyme was active on daidzin, genistin, and glycitin, inactive on esculin and apigenin-7-glucose. The EDTA and DTT as reducing agents inhibited $\beta$-glucosidase activity, but SDS and mercaptoethanol did not inhibit. Monovalent or divalent metal ions such as $MnSO_4$, $CaCl_2$, KCl, and $MgSO_4$ did not inhibited $\beta$-glucosidase activity. $CuSO_4$ and NaCl showed low inhibition, and $ZnSO_4$ inhibited 3.3 times higher than control.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.2
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• pp.117-127
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• 2008

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Cayratia japonica extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities($FSC_{50}$) of extract/fractions of Cayratia japonica were in the order: 50% ethanol extract(114.3 ${\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of some Cayratia japonica extracts in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol extract(1.21 ${\mu}g/mL$)${\mu}g/mL$). Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Cayratia japonica on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Cayratia japonica extracts suppressed photohemolysis in a concentration dependent manner($1{\sim}25{\mu}g/mL$), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect(${\tau}_{50}$, 175.05min at 25 ${\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Cayratia japonica extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments(360 nm). Two components were identified as luteolin(composition ratio, 47.50%), apigenin(52.50). TLC chromatogram of ethyl acetate fraction of Cayratia japonica extract revealed 3 bands and HPLC chromatogram showed 4 peaks, which were identified as luteolin-7-O-${\beta}$-D-glucopyranoside(composition ratio, 11.14%), apigenin-7-O-${\beta}$-D-glucuronopyranoside(15.38%), luteolin(23.55%) and apigenin(49.92%) in the order of elution time. The inhibitory effect of aglycone fraction on elastase($IC_{50},\;70.5{\mu}g/mL$) was very high. These results indicate that extract/fractions of Cayratia japonica 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. And component analysis of Cayratia japonica extract and antioxidative effects could be applicable to new cosmetics.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.1
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• pp.25-35
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• 2008

In this study, the antioxidative effects and inhibitory effects on elastase and tyrosinase of Geranium nepalense extracts were investigated. The free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities ($FSC_{50}$) of extract/fractions of Geranium nepalense were in the order: 50% ethanol extract(15.0 ${\mu}g/mL$)${\mu}g/mL$). ${\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities($OSC_{50}$) of some Geranium nepalense extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescense assay. The order of ROS scavenging activities were 50% ethanol extract($OSC_{50},\;0.23{\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Deglycosylated flavonoid fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Geranium nepalense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Geranium nepalense extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid fraction exhibited the most prominent celluar protective effect (${\tau}_{50}$, 676.7 min at 50 ${\mu}g/mL$). The inhibitory effect of aglycone fraction on tyrosinase($IC_{50}$, 70.0 ${\mu}g/mL$) and elastase ($IC_{50}$, 19.9 ${\mu}g/mL$) was very high. Aglycone fractions obtained from the deglycosylation reaction of ethyl-acetate fraction among the Geranium nepalense extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (370 nm). Two components were identified as quercetin(composition ratio, 15.3%), kaempferol(82.8%). These results indicate that extract/fractions of Geranium nepalense 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. And component analysis of Geranium nepalense extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.34 no.3
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• pp.189-200
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• 2008

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Quercus glauca extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity $(FSC_{50})$ of extract I fractions of Quercus glauca leaf was in the order: 50% ethanol extract $(12.45{\mu}g/mL)$ < ethyl acetate fraction $(10.47{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(8.57{\mu}g/mL)$. Reactive oxygen species (ROS) scavenging activities $(OSC_{50})$ of some Quercus glauca leaf extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activity was 50% ethanol extract $(OSC_{50},\;4.2{\mu}g/mL)$ < deglycosylated flavonoid aglycone fraction $(1.58{\mu}ug/mL)$ < ethyl acetate fraction $(0.66{\mu}g/mL)$. Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract / fractions of Quercus glauca leaf on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Quercus glauca leaf extracts suppressed photohemolysis in a dose dependent manner, particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect $({\tau}_{50}$, 398.67 min at $50{\mu}g/mL$). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Quercus glauca leaf extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (360 nm) as well. Two components were identified as quercetin (55.77%), and kaempferol (44.23 %). TLC chromatogram of ethyl acetate fraction of Quercus glauca leaf extracts revealed 6 bands $(QG1{\sim}QG6)$, Among them, isoquercitrin (QG3), hyperin (QG4), and rutin (QG6) were identified. The inhibitory effect of aglycone fraction on tyrosinase $(IC_{50},\;73.5{\mu}g/mL)$ and elastase $(IC_{50},\;16.2{\mu}g/mL)$ was high. These results indicate that extract / fractions of Quercus glauca 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. And component analysis of Quercus glauca leaf extract and inhibitory activity on tyeisinase and elastase of the aglycone fraction could be applicable to new functional cosmetics.