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.
Journal of the Korean Applied Science and Technology
/
v.29
no.3
/
pp.486-494
/
2012
Nano-emulsion with phase inversion temperature (PIT) emulsifying system was prepared to use rapeseed oil from originating Jeju in order to apply various cosmetic applications. Natural rape seed oil (NRSO) extraction was extracted using n-hexane as a solvent. NRSO extract showed a light yellowish color of viscous liquid as well as yield was $43{\pm}2.5%$. Acid value was $2.76{\pm}0.5$ and gravity was $0.89{\pm}0.05$. Droplet size of PIT-Yuche-NE with 20wt% of rapeseed oil was 50-120nm (average: $82{\pm}5.8nm$) and zeta potential was -29.5mV. It was thermodynamically good stable emulsion due to $(PEG)_{5-30}$fattyacidether. Some conclusions from the result of characteristic experiment were obtained as follows. First, the anti-oxidative activity was measured by free radical scavenging activity using DPPH (1,1-diphenyl-2-picrylhydrazyl radical). Anti-oxidative activity of PIT-Yuche-NE was $37.2{\pm}6.7%$ on 10mg/mL compared with PIT-Toco-NE (Natural tocopherol nano-emulsion, $28.8{\pm}6.5%$ on 10 mg/mL) and PIT-Nokcha-NE (Green tea extract nano-emulsion, $29.6{\pm}7.2%$ on 10mg/mL). Second, the collagen synthesis activity of PIT-Yuche-NE was $148{\pm}15.2%$ compared with PIT-Toco-NE (Natural tocopherol nano-emulsion, $121{\pm}13.5%$ on 10mg/mL) and PIT-Nokcha-NE (Green tea extract nano-emulsion, $95{\pm}12.7%$ on 10mg/mL). Third, the effectiveness of moisturizing activity of Yuche-CRM with Aramo-TS after 6 hours increase $47{\pm}3.9%$ (*p-value£0.05, n=7) whereas Both Toco-CRM was $30{\pm}5.2%$ (*p-value£0.05, n=7) and Nokcha-CRM was $35{\pm}4.5%$. Therefore, Yuche-CRM has higher moisturizing effect than other two creams. Finally, Nano-emulsion stabilizing rapeseed oil using PIT emulsifying system of this study can be used to apply cosmetics industry and pharmaceutical industry.
Journal of the Society of Cosmetic Scientists of Korea
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v.34
no.4
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pp.287-301
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2008
In order to develop a new depigmenting agent, extracts were obtained from 60 native plants and their antimelanogenic activities were screened by evaluating the inhibitory effect on tyrosinase which is a major enzyme responsibles for the melanin synthesis. The extracts of Trichosanthes kirilowii fruits, Phyllostachys bambusoides inner films (BIF), Clerodendrum trichotomum leaves, and Acer okamotoanum leaves showed relatively high inhibitory effect on tyrosinase and their $IC_{50}$ values were $50{\sim}100{\mu}g/mL$. The extract of BIF inhibited melanin synthesis of B16F10 melanoma cells by 52%, which was the highest among those of various extracts. Furthermore, the effect of BIF extract is 10% higher than that of arbutin (42%), a popular depigmenting agent in Korea. Ten compounds having antimelanogenic activity were isolated from the BIF extract by solvent extraction and chromatography. These compounds were identified as phenolic derivatives: SM701, SM702, SM703, and BPR211 were hydroquinone derivatives; SM707 a gallic acid derivative; SM704, SM705, SM706, SM708 and SM709 ferulic acid derivatives. The free radical scavenging activities of these compounds were measured and compared to those of hydroquinone and vitamin C. The $SC_{50}$ values scavenging 50% DPPH of SM702 and SM709 were $60{\sim}70{\mu}M$ similar to that of hydroquinone and those of SM701 and SM708 were $30{\sim}40{\mu}M$ slightly lower than that of vitamin C. These results suggest the presence of components having high antioxidant activity in the BIF extract. The SM709, identified as 1,2-O-diferulylglycerol, inhibited the activities of tyrosine hydroxylase and dopa oxidase by 18 and 60%, respectively. The SM709 also inhibited the melanin synthesis of B16F10 melanoma cells by 62% and this was the highest antimelanogenic activity among those obtained from the various purified compounds. Therefore, antimelanogenic activity of the BIF extract was concluded to be due to both inhibition of DOPA oxidase and antioxidant activity.
Kim, Taewan;Lee, Jaemin;Jeong, Gyeong Han;Kim, Tae Hoon
Food Science and Preservation
/
v.23
no.2
/
pp.283-289
/
2016
Naturally occurring antioxidants, such as polyphenols are widely found in fruits, vegetables, wines, juices, and other plant-based dietary sources and are divided into several sub classes, including phenylpropanoids, flavonoids, stilbenoids, and lignans. As part of the our ongoing search for bioactive food ingredients, the antioxidant and advanced glycation end products (AGEs) formation inhibitory activities of the methanolic extract of the aerial parts of Cirsium setidens were investigated in vitro bioassay system. The antioxidant properties were evaluated through radical scavenging assays using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) ($ABTS^+$) radicals. In addition, the activity of C. setidens against diabetes complications was also tested via AGEs formation inhibition assay. The total phenolic contents were determined using a UV-VIS spectrophotometric method. All tested samples showed a dose-dependent radical scavenging and AGEs inhibitory activities. In particular, the n-butanol (BuOH)-soluble portion showed the most potent radical scavenging activities against DPPH and $ABTS^+$ radicals with $IC_{50}$ values of $24.3{\pm}1.7$ and $25.0{\pm}3.3{\mu}g/mL$, respectively. Futhermore, the inhibition of AGEs formation by the n-BuOH-soluble portion ($IC_{50}$ value; $46.0{\pm}1.5{\mu}g/mL$) was higher than that those of the soluble portions for the other solvent. The results showed that C. setidens could be considered as an effective source of natural antioxidants and other ingredients.
Scutellaria radix (SR) has been utilized as a traditional medicine for a variety of diseases including Rheumatoid arthritis and its major flavonoids - baicalein, baicalin, and wogonin - have been reported to exert beneficial health effects, including anti-bacterial, anti-viral, anti-inflammatory, and free-radical scavenging. However, the mechanisms underlying this effect remain poorly understood. The principal objective of this study was to determine the effect of SR on osteoblast and osteoclast cells. SR extract was prepared using 70% ethanol solvent. Osteoblastic MC3T3-E1 cells and osteoclast precursor Raw 264.7 macrophage cells were utilized. SR extract increased MC3T3-E1 cell proliferation and stimulated alkaline phosphatase activity dose-dependently, 152.0% of the control at concentration $1{\mu}g/mL$. Additionally, SR extract ($1{\mu}g/mL$) stimulated Bone nodule formation activity in MC3T3-E1 cells, approximately 223.3% of the control, 20 days after the exposure. In addition, SR extract significantly reduced the number of tartrate-resistant acid phosphatase-positive (TRAP+) multinucleated cells from Raw 264.7 cells. In conclusion, SR extract stimulates the proliferation and bioactivities of boneforming osteoblasts, and inhibits the activities of bone-resorbing osteoclasts to a certain degree.
Cho, Won June;Yoon, Hee Seung;Kim, Yong Hyun;Kim, Jung Min;Yoo, Il Jae;Han, Man-Deuk;Bang, In Seok
Journal of Life Science
/
v.23
no.8
/
pp.989-997
/
2013
In this study, based on the antioxidative effects in organic solvent fractions obtained from the main methanolic extract of L. japonica, the protective cellular effects and gene expression patterns of ethyl acetate fractions on $H_2O_2$-induced Raw 264.7 cell death ($IC_{50}$) were analyzed. The antioxidant activity of the fractions measured using DPPH free radical scavenging activity increased in a dose-dependent manner, and the $ED_{50}$ exhibited the highest $39.56{\mu}g/ml$ in the ethyl acetate fraction. In addition, the ethyl acetate fractions' cell viability on $H_2O_2$-induced Raw 264.7 cell damage increased in a concentration-dependent manner, showed a visible cell survival rate of 82.49% at a concentration of $100{\mu}g/ml$. The gene expression patterns related to the ethyl acetate fractions' cytoprotective effect in $H_2O_2$-induced Raw 264.7 cell damage presented similar patterns to those of BHA. In comparative analysis for antioxidant activity-related genes affected by ethyl acetate fractions and BHA in $H_2O_2$-induced Raw 264.7 cells, both ethyl acetate fractions and BHA showed very similar gene expression patterns, but the gene expression level of the heme oxygenase 1 (Hmox1) gene making antioxidant enzymes in cells was four times higher in ethyl acetate fractions than BHA. In inflammation-related genes in $H_2O_2$induced Raw 264.7 cells, the T-box transcription factor (Tbx21) gene was expressed about two times more frequently in the ethyl acetate fraction treatment group, while it was expressed half as frequently in the BHA treatment group.
Park, Jae-Hyong;Kim, Il-Jin;Lee, Dong-Jin;Sim, Jae-Hak;Song, Min-Seop;Lee, Young-Hee;Yoo, Jung-Whan;Kim, Han-Do
Clean Technology
/
v.24
no.2
/
pp.112-118
/
2018
Thermo-responsive polymers that exhibit phase transition in response to temperature change can be used as materials for smart windows because they can control solar light transmission depending on the outside temperature. The development of thermo-responsive polymers for smart windows that can be used over a wide temperature range is desirable. To obtain high performance smart windows materials, three-dimensional thermo-responsive poly(N-isopropylacrylamide) (PNIPAm) gels were prepared by free radical polymerization from monomer N-isopropylacrylamide, N, N'-methylenebis acrylamide (MBAm) as a crosslinking agent, ammonium persulfate (APS) as a strong oxidizing agent/tetramethylene diamine as a catalyst, and a mixture of two solvents (water/glycerol). This study examined the effect of glycerol content on the lower critical solution temperature (LCST), freezing temperature and the solar light transmittance of crosslinked PNIPAm gel films. The LCST and freezing temperature of PNIPAm gel films were found to be significantly decreased from 34.3 and $6.3^{\circ}C$ to 28.2 and $-6.5^{\circ}C$ with increasing glycerol content from 0 wt% to 10 wt%, respectively. It was found that the transparent PNIPAm gel films at $25^{\circ}C$ (temperature < LCST) were converted to translucent gels at higher temperature ($45^{\circ}C$) (temperature > LCST). These results suggested that the crosslinked PNIPAm gel materials prepared in this study could have high potential for application in smart glass materials.
Journal of the Korean Society for Marine Environment & Energy
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v.8
no.2
/
pp.60-66
/
2005
When external pressure higher than osmosis pressure is reversely derived into solution, its solvent is moved into the solution having lower concentration, which is called 'reverse osmosis'. We investigated the desalination application of deep ocean water using reverse osmosis pressure of $40-70\;kgf/cm^2$ We observed how to operational factor j like flow rate, water temperature and pressure have effect on efficiency of reverse osmosis membrane and salts rejection. Fluxes of reverse osmosis membrane are directly proportional to water temperature and pressure. However, salts rejection rates are positively correlated with pressure and inversely proportional to water temperature. Separation efficiencies of osmosis membrane for major elements such as $Mg^{2+},\;Ca^{+2},\;Na^+\;and\;K^+$ are as follows in a strong electrolysis solution like seawater; $Ca^{2+},\;Mg^{2+}>K^+>Na^+$. Rejection rates of $Mg^{2+}\;and\;Ca^{2+}$ that have high electric charges are over 99% and show positively correlation with water temperature. Rejection rates of $Na^+$ having low electric charge is observed to be 98%-99%, which rates is much lower than those of $2^+$ charged ions like $Ca^{2+}\;and\;Mg^{2+}$. Ion rejection rates of boron, B, are much low because boron is present il free state or gas phase in seawater. Boron concentration in desalination water is over criteria of Korean drinking water, 0.3 mg/L. However, we could satisfied with the criteria of drinking water under the operation condition like temperature $5^{\circ}C$ and pressure $70kgf/cm^2$, using the relationship that rejection rates of boron is proportional to pressure and is inversely proportional to water temperature
Ha, Ji Hoon;Lee, Hye Mi;Kwon, Soon Sik;Kim, Hae Soo;Kim, Moon Jin;Jeon, So Ha;Jeong, Yoo Min;Hwang, Jun Pil;Park, Jong-Ho;Choi, Yung-Key;Park, Jino;Park, Soo Nam;Park, Dong-Sik
Journal of the Society of Cosmetic Scientists of Korea
/
v.39
no.4
/
pp.259-269
/
2013
In this work, licorice extracts were prepared using various extraction conditions such as extraction solvent, temperature, and time from Glycyrrhiza uralensis (G. uralensis) produced in Korea and China and Glycyrrhiza glabra (G. glabra) in Uzbekistan. The optimum extraction condition was selected from the extraction yields and antioxidative activities of extracts. Korea licorice extracts showed the highest free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activity (46.05%) under the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours. The prominent ROS (reactive oxygen species) scavenging activity using luminol-dependent chemiluminescence assay and the cellular protective effect against $^1O_2$ induced cellular membrane damage were also shown from the extracts obtained from the same condition. Especially, Korea G. uralensis extracts exhibited the higher prominent protective effect (${\tau}_{50}$ = 116.4 min) than (+)-(+)-${\alpha}$-tocopherol (${\tau}_{50}$ = 28.5 min) and the extraction yield of Korea licorice extract was 18.75%, which is 1.2 times and 2.5 times higher than that of Uzbekistan and China, respectively. These results indicate that the extraction condition of 85% ethanol at $60^{\circ}C$ for 6 hours is optimal to prepare licorice extracts, which can be applicable as anti-oxidative cosmetic materials.
Bae, Soo Jung;Song, Min Hyeon;Oh, Jung Young;Bae, Jun Tae;Kim, Jin Hwa;Lee, Geun Soo
Journal of the Society of Cosmetic Scientists of Korea
/
v.44
no.2
/
pp.117-124
/
2018
In this study, cosmetic materials were developed using a new method of making juice through the fermentation of raw natural materials with microorganisms in order to supplement the advantages and disadvantages of an organic solvent extraction method and a microbial fermentation method. The natural products were selected from two colors (red, green) of paprika known to be rich in various colors and vitamins. The microorganisms used for fermentation were fermented by inoculating paprika with lactic acid bacteria (Lactobacillus plantarum) having sugar-hydrolyzed ability. First, we investigated the changes of physiologically active substances of two kinds of paprika juice and two kinds of fermented paprika juice. Total phenols content and total flavonoids content were higher in the fermented paprika juice than in the paprika juice, and especially in the fermented red paprika juice. Free radical scavenging effect and lipid peroxidation inhibitory effect were also showed an excellent antioxidative effect on paprika fermented juice, among which the effect of red paprika fermentation juice was the highest. The expression of MMP-1 in fermented red paprika juice with high antioxidant activity was inhibited by concentration-dependent expression of MMP-1 mRNA and MMP-1 protein. In the glycation experiments with aging, the anti-glycation effect of fermented paprika juice was highly inhibited by the production of advanced glycation end-products (AGEs), which was closely related to the antioxidant effect. In addition, the activity of senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal), an indicator of cell senescence, was measured using human dermal fibroblast (HDF). The results showed that the cell senescence was inhibited when the cells were treated with fermented paprika juice. In conclusion, fermented paprika juice using lactic acid bacteria showed better antioxidative and anti-aging effects than paprika juice. Among them, fermented red paprika juice has the best antioxidant and anti-aging effect and can be applied as natural new material of antioxidant and anti-aging.
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