• Title/Summary/Keyword: Protein Fraction

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Skin Whitening and Anti-Wrinkle Effects of Extract from Jubak of Oriental Herbal Liquor (한방 발효주 주박 추출물의 미백 및 피부 주름 개선 효과)

  • Lee, Su-Min;Lee, Sang-Jin;Kwon, Yi-Young;Baek, Sang-Hoon;Kim, Jong-Sik;Sohn, Ho-Yong;Shin, Woo-Chang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1695-1700
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    • 2014
  • Oriental herbal liquor (Yakju) is a type of Korean traditional alcoholic beverage that uses Nuruk and oriental herbs for fermentation. The purpose of this study was to develop cosmetic ingredients using Jubak, which is a by-product of alcoholic fermentation of oriental herbal liquor. To investigate antioxidant, whitening, and anti-aging effects of Jubak, we prepared extract of Jubak and its solvent fractions. Ethyl acetate fraction (KSD E4-3) showed the most prominent free radical [1,1-diphenyl-2-picrylhydrazyl (DPPH)] scavenging activity ($SC_{50}$: 0.75 mg/mL). KSD E4-3 significantly inhibited in vitro mushroom tyrosinase activity ($IC_{50}$: 0.82 mg/mL) and reduced the melanin contents in mouse melanoma melanocyte, B16F10 cells. KSD E4-3 down-regulated protein expression of tyrosinase related proteins (TRP)-1, -2, which play key roles in melanogenesis. For anti-aging effects, inhibition of matrix metalloproteinase (MMPs) expression was evaluated using human keratinocyte, HaCaT cells. Treatment of HaCaT cells with KSD E4-3 reduced expression of MMP-1, -2, -9 and inhibited proteolytic activities of MMP-2, -9. These results suggest that KSD E4-3 induces down-regulation of cellular melanogenesis and protects against photoaging induced by UVB-induced damage. Thus KSD E4-3 could potentially be a valuable cosmetic ingredient.

Improvement of production of active cyclodextrin glucanotransferase by coexpression GroEL/ES chaperons in E. coli (E. coli에서 GroEL/ES chaperone 공발현에 의한 활성형 cyclodextrin glucanotransferase의 생산 증대)

  • 권미정;박소림;김병우;김성구;남수완
    • Journal of Life Science
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    • v.12 no.6
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    • pp.688-693
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    • 2002
  • Molecular chaperones prevent the misfolding of newly synthesized polypeptides in the cell. The coexpression of molecular chaperones could be expected to improve the production of soluble and active recombinant proteins. In this study, the effect of coexpression of E. coli GroEL/ES chaperone on the active production of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in E. coli was investigated. Two plasmids, pTCGT1 and pGro7 in which the cgt and the groEL/ES genes are under the control of 77 promoter and araB promoter, respectively, were co-transformed into E. coli. With a series of cultures of recombinant E. coli cells, the optimal concentrations of IPTG and L-arabinose were found be 1 mM and 0.3 mg/$m\ell$, respectively. When IPTG and L-arabinose were added at 0.8~1.0 $OD_{600}$ and 0.4~0.5 $OD_{600}$, active CGTase production was increased significantly. This coexpression condition resulted in 1.5-fold increased level of soluble CGTase (0.7~0.73 unit/$m\ell$), compared to the level of CGTase in the single expression (0.36~0.56 unit/$m\ell$). An SDS-PACE analysis revealed that about 33.6% of CGTase in the total CGTase protein was found in the soluble fraction by coexpression of GroEL/ES chaperone.

Purification of an Antibacterial Peptide from the Gills of the Pufferfish Takifugu pardalis (졸복의 아가미로부터 항균성 펩타이드의 정제)

  • Kim, Tae Young;Go, Hye-Jin;Park, Nam Gyu
    • Journal of Life Science
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    • v.27 no.1
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    • pp.50-56
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    • 2017
  • An antibacterial peptide was purified from an acidified gill extract of the pufferfish Takifugu pardalis. The acidified gill extract was put through a Sep-Pak C18 solid phase extraction cartridge using a stepwise gradient and divided into a flow-through (F.T.) and 60% methanol fraction (RM 60). Among the eluents, RM 60 had potent antibacterial activity against Bacillus subtilis KCTC 1021. RM 60 was partially purified on a cationic-exchange column (SP-5PW) by a linear gradient, and the antibacterial peptide was then further purified, using a series of cationic-exchange and $C_{18}$ reversed-phase HPLC columns. For characterization of the purified peptide, its molecular weight and amino acid sequence were analyzed by MALDI-TOF MS and Edman degradation. The molecular weight of the peptide was about 1171.6 Da. The amino acid sequence of the peptide was partially determined as: STKEKAPRKQ. A comparison of the N-terminal amino acid sequence of the purified peptide with that of other known polypeptides revealed high homology with the N-terminus of the histone H3 protein, which belongs to the histone H3 family. Thus, this peptide was designated as a puffer fish gill (PFG)-related antimicrobial peptide. This is the report to describe an antimicrobial function for the N-terminus of histone H3 of an animal species. The findings suggest that this peptide plays a significant role in the innate defense system of the pufferfish.

Isolation and Characterization of MMP-1 Inhibitor Peptide from Crataegus pinnatifida Bunge in Fibroblast Cell Line HS68 Cells (아가위(Crataegus pinnatifida Bunge)로 부터 HS 68세포의 MMP-1에 대한 저해활성 물질의 분리)

  • Lee, Se-Young;Chun, Hyug;Cho, Hong-Yun;An, Jeung-Hee
    • Applied Biological Chemistry
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    • v.46 no.1
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    • pp.60-65
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    • 2003
  • MMP-1 inhibitory compounds were isolated from 120 Korean traditional edible plants. UP- 1 activity significantly increased linearly with increasing UVB dose in normal human foreskin fibroblast HS68 cell, showing maximum activity at approximately 35 $mJ/cm^2$, whereas in HaCaT cell, normal human keratinocyte, no increase was observed. Maximum secretion of MMP-1 after UVB treatment occurred around 36-48 k after treatment. MMP-1 inhibitory compound isolated from cold-water fraction of Cataegus pinnatifida Bunge showed the mort potent activity. The MMP-1 inhibitory compound was deduced as a peptide based on the fact that pronase digestion decreased the activity whereas periodate oxidation did not. The most potent UP- 1-inhibitory protein, CP-2Va-2, showing an activity of 88.5% against MMP-1, was isolated through sequential column chromatography on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M, and Bio-Gel P-30. Molecular weight of CP-2Va-2 determined through high performance liquid chromatography and SDS PACE was 19 and 20 kDa. respectively, signifying a monomeric structure.

Characterization of Mitochondrial Plasmids from Pleurotus spp. (Pleurotus속 균주들의 미토콘드리아 플라스미드 특성)

  • 김은경;구용범;차동렬;하영칠;노정혜
    • Korean Journal of Microbiology
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    • v.31 no.2
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    • pp.141-147
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    • 1993
  • Plasmid DNAs were detected from the mitochondrial fraction of four strains of whiterot fungus, Pleurotus ostreatus. The size of the plasmids were 10.2 and 7.2 kb in strain NFFA 2, 10.2 kb in NFFA 4001, 11.2 kb in NFFA 4501, and 10.2 and 11.2 kb in KFCC 11635. The two strains,NFFA 2ml and NFFA 2m2, which are mutant derivatives of NFFA 2, did not contain any plasmids. The cleavage by proteinase K indicated that these plasmids have DNA ends associated with proteins. In digestion with proteinase K all the plasm ids remained resistant to lambda exonuclease which hydrolyzes DNA from 5' ends and were sensitive to exonuclease III which hydrolyzes DNA from 3' ends. This suggests that the plasmids are linear double-stranded DNA and the terminal proteins are covalently linked to 5' ends of plasm ids. In order to find relationship between these plasmids, hybridization of plasm ids by each separate plasmid DNA was done. The result indicated that the plasmids can be classified into at least 3 groups. Plasmids of group I were present in all the P ostreatus. More mitochondrial plasmids were detected in P cornucopiae. P ,florida, P pulmonarius, P sajor-caju, and P spodoleucus. The size of plasmids ranged between 7.2 kb and 14 kb. All the species except P cornucopiae contained plasmids of approximately 10 kb which hybridized with the 10.2 kb plasmid (group I) of P ostreatus NFFA 2.

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Hydrolysis of Agricultural Residues and Kraft Pulps by Xylanolytic Enzymes from Alkaliphilic Bacillus sp. Strain BK

  • Kaewintajuk Kusuma;Chon Gil-Hyong;Lee Jin-Sang;Kongkiattikajorn Jirasak;Ratanakhanokchai Khanok;Kyu Khin Lay;Lee John-Hwa;Roh Min-Suk;Choi Yun-Young;Park Hyun;Lee Yun-Sik
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1255-1261
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    • 2006
  • An alkaliphilic bacterium, Bacillus sp. strain BK, was found to produce extracellular cellulase-free xylanolytic enzymes with xylan-binding activity. Since the pellet-bound xylanase is eluted with 2% TEA from the pellet of the culture, they contain a xylan-binding region that is stronger than the xylan-binding xylanase of the extracellular enzyme. The xylanases had a different molecular weight and xylan-binding ability. The enzyme activity of xylanase in the extracellular fraction was 6 times higher than in the pellet-bound enzyme. Among the enzymes, xylanase had the highest enzyme activity. When Bacillus sp. strain BK was grown in pH 10.5 alkaline medium containing xylan as the sole carbon source, the bacterium produced xylanase, arabinofuranosidase, acetyl esterase, and $\beta$-xylosidase with specific activities of 1.23, 0.11, 0.06, and 0.04 unit per mg of protein, respectively. However, there was no cellulase activity detected in the crude enzyme preparation. The hydrolysis of agricultural residues and kraft pulps by the xylanolytic enzymes was examined at 50$^{\circ}C$ and pH 7.0. The rate of xylan hydrolysis in com hull was higher than those of sugarcane bagasse, rice straw, com cop, rice husk, and rice bran. In contrast, the rate of xylan hydrolysis in sugarcane pulp was 2.01 and 3.52 times higher than those of eucalyptus and pine pulp, respectively. In conclusion, this enzyme can be used to hydrolyze xylan in agricultural residues and kraft pulps to breach and regenerate paper from recycled environmental resources.

Anticancer and Immunopotentiating Activities of Crude Polysaccharides from Pleurotus nebrodensis on Mouse Sarcoma 180

  • Cha, Youn Jeong;Alam, Nuhu;Lee, Jae Seong;Lee, Kyung Rim;Shim, Mi Ja;Lee, Min Woong;Kim, Hye Young;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Tae Soo
    • Mycobiology
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    • v.40 no.4
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    • pp.236-243
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    • 2012
  • Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ${\beta}$-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of $10{\sim}2,000{\mu}g/mL$. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of $50{\mu}g/mL$, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at $200{\mu}g/mL$. Maximum production of nitric oxide ($14.3{\mu}M$) was recorded in the Fr. NaCl fraction at $200{\mu}g/mL$. Production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-${\alpha}$, IL-$1{\beta}$, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

The Isolation and Pyrolysis of the Brown Pigmented Macromolecule from the Cured Leaf Tobacco (잎담배 성분중 갈색고분자 물질의 분리정제 및 열분해에 관한 연구)

  • Chae, Quae;Park, Ji-Chang
    • Journal of the Korean Society of Tobacco Science
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    • v.2 no.1
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    • pp.1-7
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    • 1980
  • Gel filtration column chromatography (Sephadex G-75), dialysis an d Brushite column chromatography were carried out to separate the brown pigmented macromolecule from water extracts of the cured leaf tobaccos. The two distinct macromolecules having different molecular weight were separated by the Sephadex column chromatography. Brushite also separated two different species of macromolecules which might have different electronic structures. According to the enzymatic degradation of protein in Burley and Hicks, chymotrypsin showed the best degradation ratio, ie., 16-30% in Burley and 38-57% in Hicks. Similar effect was observed with pepsin. However, very low effect of degradation was revealed with trypsin. The sample treated with the proteolytic enzymes revealed the disappearance of the first peak and the slight decrease of the 2nd peak height in the separation profile of Sephadex. After dialysis, the brown pigmented macromolecule was pyrolyzed at $300^{\circ}C$ and the strongly fluorescent components not identified before pyrolysis were detected with TLC separation. Absorption spectrum of these fluorescent compounds was monitored in benzene and the absorption maxima at 265nm and 275 nm were obtained. Considering absorption maxima and shape of the spectrum, those fluorescent compounds seem to be PAH derivatives.

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Effects of Dietary n-3 Highly Unsaturated Fatty Acids on Growth and Biochemical Changes in Korean Rockfish Sebastes schlegeli III. Changes of Body Compositions with Starvation (사료의 n-3계 고도불포화지방산 함량에 따른 조피볼락 Sebastes schlegeli의 성장 및 생화학적 변화 III. 절식시 체조성의 변화)

  • LEE Sang-Min;HUR Sung Bum
    • Journal of Aquaculture
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    • v.6 no.3
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    • pp.199-211
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    • 1993
  • In order to elucidate the effect of dietary n-3 highly unsaturated fatty acids (n-3HUFA) on the changes of body weight and chemical compositions in the Korean rockfish during starvation, the fish were not fed for 9 weeks after fed different levels $(0\~1.5\%)$ of n-3 HUFA for 10 weeks. The higher level of n-3HUF A was contained in the diets, the slower body weight loss was resulted (P< 0.05). The decreasing rates of the body nutrients of the fish were significantly higher in the fish fed n-3HUF A deficient diets than those of the fish fed n-3 HUF A sufficient diets. Protein and lipid contents of the whole body were decreased with starvation whereas moisture content was increased. Decrease of lipid was mainly due to the decrease of nonpolar lipid. Amounts of polar lipid in the whole body were almost constant throughout the starvation, meaning not being affected by dietary n-3HUF A levels. Percentage of 22: 6n-3 was increased in the polar lipid fraction, but monoenic acids (16:1, 18:1), n-3 series (18:3, 18 4, 20:4) and n-6 series (20:2, 22:4, 22:5) were decreased with starvation. Fatty acid compositions of nonpolar lipid were not changed with starvation. These results suggest that all fatty acids of nonpolar lipid are equally utilized for energy during starvation.

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Taxifolin Inhibited the Nitric Oxide Production and Expression of Pro-inflammatory Cytokine mRNA in Lipopolysaccharide-stimulated RAW264.7 Cells

  • Rhee, Man-Hee;Endale, Mehari;Kamruzzaman, SM;Lee, Whi-Min;Park, Hwa-Jin;Yoo, Myung-Jo;Cho, Jae-Youl
    • Biomedical Science Letters
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    • v.14 no.3
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    • pp.147-155
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    • 2008
  • In previous works, we found that solvent extract of Opuntia humifusa Raf., a member of the lactaceae family, displayed potent anti-oxidative and anti-inflammatory activities. Thus, all solvent fractions, except for the water layer, showed potent scavenging effects. According to activity-guided fractionation, one of active radical scavenging principles in the ethyl acetate fraction was found to be taxifolin. In this study, we investigated whether taxifolin showed anti-oxidative activity. In addition, taxifolin modulated nitric oxide (NO) release and the expression of pro-inflammatory cytokine mRNA such as interleukin-$1{\beta}$ (IL-$1{\beta}$), IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), and TNF-${\alpha}$. Taxifolin showed potent anti-oxidant activity with the $IC_{50}\;of\;8.5{\pm}1.4\;and\;9.3{\pm}1.0{\mu}M$ using xanthine/xanthine oxidase (XO) assay and 2,2-Diphenyl-lpicrylhydrazyl radical (DPPH) assay, respectively. We next determined the role of taxifolin on the immunomodulating activity using murine macrophage cell line RAW264.7 cells. Taxifolin dose-dependently inhibited NO production in lipopolysaccharide (LPS)-activated RAW264.7. It also significantly blocked the expression of inducible NO synthase (iNOS) mRNA in the LPS-stimulated RAW264.7 cells. In addition, taxifolin potently suppressed the expression of IL-$1{\beta}$, IL-6 and GM-CSF mRNA in LPS-activated RAW264.7 cells, but not that of TNF-${\alpha}$ Moreover, taxifolin significantly inhibited the transcriptional activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) and activator protein -1 (AP-1). These results suggest that taxifolin may downregulate inflammatory iNOS, IL-$1{\beta}$, IL-6 and GM-CSF gene expressions through inhibition of NF-K and AP-1 activation in LPS-stimulated RAW264.7 cells.

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