• Title/Summary/Keyword: molecular weight control

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Cloning of $\beta$-Glucosidase Gene from Streptomyces coelicolor A3(2) and Characterization of the Recombinant $\beta$-Glucosidase Expressed in Escherichia coli (Streptomyces coelicolor A3(2)로 부터 $\beta$-Glucosidase 유전자 클로닝 및 재조합 효소의 특성)

  • Kim, Jae-Young;Kim, Bong-Kyu;Yi, Yong-Sub;Kang, Chang-Soo;Ahn, Joong-Hoon;Lim, Yoong-Ho
    • 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.

Phytoremediation by Persicaria thunbergii (고마리를 사용한 중금속 처리)

  • 강경홍;김인성
    • Journal of Korea Soil Environment Society
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    • v.5 no.1
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    • pp.33-43
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    • 2000
  • For the consideration of phytoremediation, TEX>$Cd^{2+}$ and Pb$^{2+}$ were analysed in the soil of the habitats and the leaf stem and root of Persicaria thunbergii in the different localities of Bong-Dong river In the soil and plant samples of research areas, TEX>$Cd^{2+}$ was not detected but, $Pb^{2+}$ detected as follows; about 7.5~15.5$mu\textrm{g}$/g in the soil of habitats, about 11.7~18.4 $mu\textrm{g}$/g in the leaf, about 7.~15.5$mu\textrm{g}$/g in the stem and about 89.1~193.6$\mu\textrm{g}$/g in the root of P. thunebrgii and the correlation coefficient value between the $Pb^{2+}$ contents in soil and P. thunbergii was 0.814(>t12, 0.01). After P thunbergii was treated with Cd(NO$_3$)$_2$and Pb(NO$_3$)$_2$of 5 and 10mM, the bioaccumulation of TEX>$Cd^{2+}$ and $Pb^{2+}$ in the leaf of plant, the remaining mass of heavy metals and the variation of pH in the soil, and the increasing rate(%) of phytochelatin in plant were examined. The concentrations of TEX>$Cd^{2+}$and $Pb^{2+}$ in the leaf as follows, in the case of TEX>$Cd^{2+}$ about 0.82~2.79$\mu\textrm{g}$/g and in $Pb^{2+}$, about 2.87~8.08$\mu\textrm{g}$/g. The remaining mass of heavy metals and the variation of pH in the cultured soil decreased as follows; about 77.1% and pH6.39 in TEX>$Cd^{2+}$ 5mM, about 90.2% and pH5.79 in TEX>$Cd^{2+}$ 10mM, about 81.1% and pH6.00 in $Pb^{2+}$ 5mM and about 85.7% and pH5.80 in Pb$^{2+}$ 10mM. The phytochelatin were increased in plant samples treated with 10mM Cd(NO$_3$)$_2$and Pb(NO$_3$)$_2$as follows; about 259% by TEX>$Cd^{2+}$ and about 305% by Pb$^{2+}$ be compared with control. and the molecular weight(da) of these phytochelatins were estimated about 4,300~8,600da in the case of the treatment of TEX>$Cd^{2+}$ and about 3,200~9,700 in $Pb^{2+}$./TEX>.

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ELISA of paragonimiasis in cat by crude and purified antigens of Paragonimus westermani (폐흡충(肺吸蟲)의 조(粗)항원과 정제(精製)항원에 의한 폐흡충(肺吸蟲)감염 고양이혈청의 면역효소반응(免疫酵素反應))

  • Lee, Ok-Ran;Chang, Jae-Kyung
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.187-193
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    • 1986
  • Enzyme-linked immunosorbent assay(ELISA) using crude and affinity-purified antigens of adult worms of Paragonimus westermani was performed for infected cat sera with different worm burden, from preinfection to 18th week after infection. Crude antigen was used with supernatant of homogenated worms by freezing-thawing method, and the supernate was centrifuged for 1 hour at 10,000 rpm at $4^{\circ}C$. Affinity-purified antigen(antibody-bound antigen) was prepared from fractions(bound and unbound) of crude antigen by affinity chromatography on CNBr-activated sepharose 4B, and IgG as a ligand was prepared from paragonimiasis cat serum(6 months infected) obtained by ammonium sulfate ($40%{\sim}45%$ saturated) precipitation method. By SDS-PAGE, crude antigen showed 22 polypeptide fractions while purified antigen showed 4 fractions: 36, 400, 34, 700, 27, 600 and 11, 500 in molecular weights. All cats were divided into five groups($G_1-G_5$) by different worm burdens. The mean of recovered worms(${\pm}SD$) and the number of cats in each group are as follows: $G_1$, 2 worms(0) and 4 cats; $G_2$, 4.75 (${\pm}0.66$) and eight; $G_3$, 10.75(${\pm}1.92$) and four; $G_4$, 23.20(${\pm}3.43$) and five; $G_5$, 48(${\pm}12.63$) and five cats. The results were summarized as follows: 1. The antibody levels(OD value) increased by worm burden in $G_1$ to $G_4$ generally. However, individual antibody levels were not exactly related with worm burden in all groups, especially there was a wide difference in $G_4$ and $G_5$. These results suggested that the worm burden in $G_4$ (about $20{\sim}30$ worms) is enough to produce antibody maximum in cats of $2{\sim}3kg$ weight. 2. The antibody levels increased significantly(p<0.05) compared to control sera at the 3rd week in $G_1$ and $G_2$, at the 2nd week in $G_3$, and at the 1st week in $G_4$ and $G_5$. Especially in the 4th week, OD value increased more in $G_1$(p<0.01) and in $G_2$ to $G_5$(p<0.001). In the pattern of antibody levels by ELISA in each group, OD in $G_1$ increased to the 18th week continuously, in $G_2$ OD was maintained same after the 16th week, but in $G_3$ it decreased after the 16th week, and it was maintained same in $G_4$ and $G_5$ after the 14th week. 3. The antibody levels by ELISA with the affinity-purified antigen were higher than those with crude antigen in all groups generally. Especially, the difference of OD values between two antigens was larger from the 4th to the 10th week. In $G_1$ and $G_2$ OD with purified antigen was higher than that with crude one to the 18th week. It was also higher in $G_3$ than that with crude antigen to the 16th week and OD of $G_4$ and $G_5$ were higher before the 14th week than that with crude antigen, however became lower at the 16th week. Consequently, the antibody level in ELISA with affinity-purified antigen was more sensitive at the early weeks after infection and in light infection groups than that with crude antigen.

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Pervaporation Separation of Ethanol Aqueous Solution through Carbonate-type Polyurethane Membrane II. The Effect of Pendent Anionic Group (카보네이트형 폴리우레탄막을 이용한 에탄올 수용액의 투과증발분리 II. 음이온성기에 의한 영향)

  • Han, In Ki;Oh, Boo Keum;Lee, Young Moo;Noh, Si Tae
    • Applied Chemistry for Engineering
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    • v.3 no.4
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    • pp.595-604
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    • 1992
  • Carbonate-type polyurethane resins containing anionic moieties were systhesized from NCO-terminated prepolymer method. Membranes were manufactured from the polymer solution and the separation of aqueous ethanol solution was investigated. To enhance the property of urethane resin, carbonate-type polyol(PTMCG) was used. ${\alpha}^{\prime},{\alpha}^{{\prime}{\prime}}$-dimethylolpropionic acid was used as a chain extender to increase the hydrophilicily of the urethane membrane. The ionization of the pendent carboxylic groups in urethane resin was carried out using trimthylamine. To confirm the formation of anionic groups in urethane resin, IR spectra of model compounds were compared with those of urethane resins. It was confirmed that the concentration of hard segment and hydrogen bond contributed to the property of the concentration of hard segment and hydrogen bond contributed to the property of urethane resin in which the mole ratio of chain extender and polyol was from 3:1 to urethane resin in which the mole ratio of chain extender and polyol was from 3:1 to 5:1. The carbonate-type polyurethane containing pendent carboxylic grop(PU) had Tg of around-$25^{\circ}C$ and Tm, $45^{\circ}C$ measured by DSC. Transition temperatures of one containing pendent anionic group(APU) prepared from the ionization of PU shifted to $8{\sim}10^{\circ}C$ lower temperature region than those of PU. Pervaporation membrane was prepared through the casting method. N, N-dimethylformamide (DMF) were used as a solvent and hexamethylene diisocyanate(HMDl) as a crosslinking agent. Swelling degree increased with ethanol concentration in mixure and the control of the swelling degree of the membrane could be achieved by crossliking. The results of pervaporation were as follows : separation factor, 2.3~9.8 ; flux, $27{\sim}79.5g/m^2hr$. Pervaporation separation capacity could be enhanced by reducing the molecular weight of polyol from 2,000 to 1,000.

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Anti-aging Effects of the Extracts from Leaf. Stem, Fruit and Seed of Yew (Taxus cuspidata Sieb) by Solvent Extraction Method (용매추출법에 의한 주목의 잎, 줄기, 과실 추출물의 항 노화 효과)

  • Kim, In-Young;Jung, Sung-Won;Ryoo, Hee-Chang;Zhoh, Choon-Koo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.211-219
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    • 2004
  • Yew (Taxus cuspidata Sieb.) chose that grow as medicine, food, decorative plant in Korea's Kyong-Gi province surroundings. Extracts of yew extracted leaf of 250g and stems of 300g with 1,3-butylene glycol (l,3-BG), propylene glycol (PG) and water. As results, external appearance of leaf extract of yew was slightly brown clear extract. The pH was 5.3${\pm}$0.5, and specific gravity was 1.012${\pm}$0.05, and refractive index was l.375${\pm}$0.05. Also, appearance of stem's extract was slightly brown clear extract, and the pH was 5.4${\pm}$0.5, and specific gravity was 1.016${\pm}$0.05, and refractive index was 1.358${\pm}$0.05. Oil of yew separated from seeds, and extracted polysaccharide high purity from fruits. As a result, specific gravity of oil was 0.987, and obtained 40.0% of yield. Total polyphenols amount of yew extract is detected 0.563% in leaves, 0.325% in stems, whereas total tannins amount contained 0.054% and 0.037% each in leaves and stems. As effect in cosmetics, the anti-oxidative effect by DPPH method is 75.0% in leaves, and stems was 64.0%. Collagen synthesis rate was shown high activity by 54.16% in stem's extract, 33.18% in leaves' extract. Also, PPE-inhibitory activities were 13.7% and 23.5% each in leaves and stems. Anti-inflammatory effect of yew seed oil displayed superior effect of 41% than control. Polysaccharide's molecular weight that is gotten from fruits was 5${\times}$10$^4$-3${\times}$10$\^$5/ dalton, and got 20.0${\pm}$5% of yield.

A Study on the Effects of Taxus Extracts in Cosmetic Indurstry (화장품산업에서 주목추출물의 효능에 관한 연구)

  • 김인영;이계종;정성원;이주동;유희창;조춘구
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.28 no.1
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    • pp.80-98
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    • 2002
  • Taxus cuspidata Sieb selected cultivation as drug, food and decorative plant in Kyong-gi province in Korea. As a manufacturing method, there were extracted from 250g of dried-leaf and 300g of dried-stem with each 200g of BG, PG and water (to 100) mixing for 72 hour at 50$\pm$5$\^{C}$ and then they were filtered by 400-mesh filter. Appearance of extract of leaves was slight brown, pH=5.3$\pm$0.5, gravity was 1.012$\pm$0.05, and a reflective index was 1.375$\pm$0.05. And appearance of extract of stems was slightly dark brown, pH=5.4$\pm$0.5, gravity was 1.016$\pm$0.05, and a reflective index was 1.358$\pm$0.05. It was extracted oil from Taxus seed. Gravity was 0.922$\pm$0.05 and it should be obtained the 27.0$\pm$0.5% of yield. The molecular weight of polysaccharide was about 50,000 to 300,000 dalton and contained 5.0$\pm$1.2% of yield from Taxus fruit. The determinations of total polyphenols in measuring spectropotometer got 0.563% in leaves, and 0.325% in stems, whereas the quantitives of total tannins got 0.054% and 0.037%, respectively. As the effects in Cosmetics by DPPH-method, the antioxidative activities were very strong that the inhibitory ratio showed 75% in leaves and 64% in stems compared with 52% in greentea extract. These are more effective than other plant extracts. The increasing ratio of collagen synthesis rate on the activating fibroblast for extracts of Taxus cuspidata Sieb showed 54.16% (stems) and 33.18% (leaves), To improve the skin elasticity, PPE(porcine pancreatic elastase)-inhibitory activities were strongly effective as 13,7% (stems), 23.5% (leaves) and 66%(seed). Anti-inflammatory acitvity of seed oil was very the above 41% stronger than SG was 24% of anti-Inflammatory as a control sample.

Effects of Corni Fructus on Testosterone-induced Benign Prostatic Hyperplasia in Sprague Dawley Rats (Sprague Dawley 흰쥐에서 테스토스테론에 의하여 유발된 전립선 비대증에 미치는 산수유 추출물의 영향)

  • Kwon, Da He;Hwangbo, Hyun;Choi, Eun Ok;Kim, Min Yeong;Ji, Seon Yeong;Kim, Kyung-Il;Park, No-Jin;Kim, Sung Ok;Hong, Su-Hyun;Park, Cheo;Hwang, Hye-Jin;Jeong, Ji-Suk;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1507-1515
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    • 2018
  • Benign prostatic hyperplasia (BPH) is characterized by prostatic hypertrophy mainly in the elderly. Corni Fructus is reportedly effective in the prevention and treatment of various diseases, but its efficacy on BPH has not been previously studied. In the present study, we investigated whether or not a Corni Fructus water extract (CF) could prevent testosterone-induced prostatic hyperplasia in rats. To induce BPH, castrated rats were subcutaneously injected with testosterone propionate (TP). CF was administered daily by oral gavage, along with the TP injections, and finasteride, a selective inhibitor of $5{\alpha}$-reductase type 2, was used as a positive control. The results show that CF significantly reduces prostate weight and histopathologic changes while also decreasing levels of serum dihydrotestosterone, similar to the finasteride-treated group. CF also suppresses TP-induced $5{\alpha}$-reductase expression and concentration in prostate tissue and serum, respectively. Furthermore, CF markedly inhibited TP-induced expression of the androgen receptor (AR) and the steroid receptor coactivator 1, an AR coactivator, which was associated with a decrease in prostate-specific antigen levels in both serum and prostate tissue. In conclusion, the results of this study indicate that CF weakens BPH status by inactivation of $5{\alpha}$-reductase and AR.