• Title/Summary/Keyword: 단백질 추출

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Hypolipidemic Effects of Glycoprotein Isolated from Ficus Carica Linnoeus in Mice (무화과 당단백질의 혈중지질 저하 효과)

  • Lim, Kye-Taek;Lee, Sei-Jung;Ko, Jeong-Hyeon;Oh, Phil-Sun
    • Korean Journal of Food Science and Technology
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    • v.37 no.4
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    • pp.624-630
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    • 2005
  • Glycoprotein (60 kDa) isolated from Ficus Carica Linnoeus (FCL glycoprotein) was examined by evaluating its hypolipidemic effects on plasma cholesterol levels and hepatic detoxicant enzyme activities in ICR mice. FCL glycoprotein $(100{\mu}g/mL)$ had strong scavenging activities (38%) against lipid peroxyl radicals. When mice were treated with Triton WR-1339 (400 mg/kg), levels of total cholesterol (TC) and low-density lipoprotein (LDL)-cholesterol in plasma significantly increased by 53.9 and 47.5 mg/dL, respectively, compared to the control, whereas, when pretreated with FCL glycoprotein $(100{\mu}g/mL)$, decreased remarkably by 55.4, and 47,0 mg/dL, compared to Triton WR-1339 treatment alone. Interestingly, high-density lipoprotein (HDL)-cholesterol level did not change. Body and liver weights did not change significantly after Triton WR-1339 treatment in presence of FCL glycoprotein. FCL glycoprotein $(100{\mu}g/mL)$ stimulated activities of antioxidative detoxicant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), whereas GPx activity significantly increased compared to the control. These results suggest FCL glycoprotein has abilities to scavenge lipid peroxyl radicals, lower plasma lipid levels, and stimulate detoxicant enzyme activity in mouse liver.

Purification and Characterization of Protease Produced by Aspergillus wentti Isolated from Korean Traditional Meju (한국 전통 메주 유래의 Aspergillus wentti가 생성하는 Protease 의 정제 및 특성)

  • Lim, Seong-Il
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.161-167
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    • 2000
  • The protease produced by a newly isolated Aspergillus wentti from Korean traditional Meju was purified and characterized. The optimal medium composition and culture conditions for maximum protease production were ; bran :1% glucose solution =1 : 1, pH 9.0, $30^{\circ}C$, and 4 days of fermentation. Protease was purified by QAE-Sephadex, SP-Sephadex ion exchange chromatography and Sephadex G-100 chromatography. The specific activity and the purification fold of the purified enzyme were 213 unit/mg protein and 27.3, respectively. The molecular weight of purified protease was found to be 32 kDa by SDS-PAGE. Km and Vmax value's for hammastein milk casein were $3.049{\times}10^{-4}\;M\;and\;151.1\;{\mu}g/min$, respectively. Kinetic parameters showed that the enzyme has higher affinity to casein than isolated soybean protein, hemoglobin and bovine serum albumin. Optimal pH and temperature for reaction of the purified enzyme were 9.0 and $50^{\circ}C$, respectively. The enzyme was stable at pH 4.0-11.0, below $40^{\circ}C$, and the activity was not stimulated by metal ions. 1mM phenylmethylsulfonyl fluoride inhibited the enzyme activity by 98.5%. It means that the enzyme is one of serine protease.

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Reinforcement of Antioxidative Potentials by Korean Traditional Prescriptions on Mouse Plasma and Liver (전통 한약 탕제 투여에 의한 혈장 및 간 조직의 항산화력 증강 효과)

  • Hong, Seong-Gil;Lee, Mi-Young;Yoon, Yoo-Sik;Kang, Bong-Joo;Kim, Dae-Won;Cho, Dong-Wuk
    • Korean Journal of Food Science and Technology
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    • v.31 no.6
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    • pp.1661-1666
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    • 1999
  • Yungmijihwgang-Won, Yollyunggobon-Dan and Palmi-Hwan, Korea traditional prescriptions composed of oriental medical herbs, have been used successfully to improve human health and regimen. This study was designed to examine the mechanism of healthful effects of the Korea traditional prescriptions through its antioxidative potentials. Using in vitro antioxidative activity assay system such as DPPH radical quenching assay, superoxide anion radical scavenging assay and inhibition of TBARS production, three Korea traditional prescriptions were observed to have nearly the same antioxidative potentials as ascorbic acid, a well-known strong water-soluble antioxidant. Moreover, we observed reinforced antioxidative effects of these drugs in liver from mouse fed these drugs with 4 weeks. When liver homogenate was incubated with 2.2'-azobis(amidinopropane) dihydrochloride(AAPH), as a free radical initiator, we observed that oxidative damages were decreased and antioxidative potentials were increased in liver homogenate treated these drugs. However, enzymatic antioxidative system as superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase was not affected by drug administration.

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Anti-Oxidative and Anti-Inflammatory Effects of the Solvent Fraction from Theobroma cacao L. Extract (카카오(Theobroma cacao L.) 용매 분획물의 항산화 및 항염증 효과)

  • Kim, Young-Sun;Lee, Jin-Young;Cho, Young-Je;An, Bong-Jeun
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1330-1338
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    • 2012
  • Solvent extracts of Theobroma cacao L. (TCL) were investigated for anti-oxidative and anti-inflammatory effects in order to consider TCL as a functional ingredient for cosmetic products. TCL(A) extract was fractioned according to polarity with $CHCl_3$, EtOAc, n-BuOH, and water. Following TCL(A) fractionation, the electron-donating ability of the n-BuOH and EtOAc solvent fractions (each 100 ${\mu}g/ml$) was about 76.2% and 53.9%, respectively. The superoxide anion radical inhibitory effect of the n-BuOH and EtOAc solvent fractions (each 50 ${\mu}g/ml$) was about 76.09% and 51.4%, respectively. Results of lipid oxidation showed that $Fe^{2+}$ had a greater chelating effect than $Cu^{2+}$. The $Fe^{2+}$ chelating effect of the EtOAc solvent fraction (50 ${\mu}g/ml$) was about 64%. Hyaluronidase inhibition related to the anti-inflammatory effect was 53.0% with EtOAc at 100 ${\mu}g/ml$, while the lipoxygenase inhibitory effect was about 51.32% at 10 ${\mu}g/ml$. The anti-inflammatory activity in the EtOAc fraction inhibited the generation of nitric oxide. Results also showed that iNOS protein expression increased in RAW264.7 cells. In contrast, at 100 ${\mu}g/ml$ EtOAc, iNOS and COX-2 protein expression significantly decreased in LPS-stimulated RAW 264.7 cells.

Apoptotic Effects of Resveratrol via mTOR and COX-2 Signal Pathways in MCF-7 Breast Cancer Cells (MCF-7 유방암 세포에서 mTOR-COX-2 신호경로를 통한 resveratrol의 apoptosis 효과)

  • Lee, Sol-Hwa;Lee, Hye-Yeon;Park, Song-Yi;Park, Ock-Jin;Kim, Young-Min
    • Journal of Life Science
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    • v.21 no.9
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    • pp.1288-1294
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    • 2011
  • Resveratrol, a kind of phytochemical, is presented in grape skins. Resveratorl exerts antiproliferative, anti-cancer and pro-apoptotic activities in cancer cells. Mammalian target of rapamycin (mTOR) is a critical regulator of cellular growth and proliferation, and it is known to be a strategic target for anti-cancer therapeutic uses. mTOR is a major downstream of the PI3K/Akt pathway, which is activated in various cancer cells. It also plays an important role in the survival, proliferation and angiogenesis of cells. Cyclooxygenase-2 (COX-2) is an important protein that mediates inflammatory processes. It plays an important role in various tumors by affecting cell proliferation, mitosis, apoptosis and angiogenesis. In this study, we have investigated the effects of resveratrol on apoptosis through mTOR and COX-2 expression in MCF-7 breast cancer cells. The treatment of resveratrol with different concentrations inhibited proliferation of MCF-7. The data showed that resveratrol induced apoptotic cell death of cancer cells and decreased mTOR and COX-2 expression. These results suggest that resveratrol induces apoptosis of MCF-7 breast cancer cells by inhibiting mTOR and COX-2 expression.

In vitro Inhibitory Effect of Aged Black Garlic Extract with Antioxidant Activity on MMP-2 and MMP-9 Related to Metastasis (In vitro에서 항산화 효능이 있는 흑마늘 추출물의 MMP-2 및 MMP-9에 대한 활성 억제효과)

  • Lee, Soo-Jin;Nam, Hyang;Kim, Moon-Moo;Jang, Ho-Jung;Park, Jung-Ae;Kim, Byung-Woo;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.20 no.5
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    • pp.760-767
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    • 2010
  • The oxidative damage of lipids, protein, and DNA is known to be involved in not only chronic inflammations such as arthritis, hepatitis, nephritis, gastritis, colitis, and periodontitis but also metastasis. It has given impetus to searching for natural compounds without toxicity, which prevent the development of these diseases. The direct scavenging effects of aged black garlic extract (ABGE) were evaluated in vitro on DPPH radical, hydroxyl radical, hydrogen peroxide, and genomic DNA damage related to oxidative stress. Furthermore, its antioxidant effect on lipid peroxidation was investigated in human fibrosarcoma cells (HT1080), which were exposed to the hydroxyl radical generated by the Fenton reaction. It was observed that ABGE exhibited a greater inhibitory effect on hydrogen peroxide than other reactive oxygen species, and also blocked DNA oxidation and lipid peroxidation induced by the hydroxyl radical. The oxidative stress in live cells was also inhibited in the presence of ABGE. In addition, its inhibitory effects on the activity and expression of MMP-2 and -9 related to metastasis were determined using gelatin zymography and western blot. The data showed that it inhibited MMP-2 and -9 in PMA-stimulated HT1080 cells. Therefore, these results suggest that ABGE show potential as an excellent agent for prevention of metastasis related to oxidative stress.

Ginsenoside Rg3 Induces Apoptosis in B16F10 Melanoma Cells (ginsenoside Rg3에 의한 B16F10 흑색종 세포의 세포사멸 유도)

  • Lee, Seul Gi;Kim, Byung Soo;Nam, Ju-Ock
    • Journal of Life Science
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    • v.24 no.9
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    • pp.1001-1005
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    • 2014
  • Ginsenoside Rg3 is one of the active ingredients extracted from red ginseng, and it is an effective chemical component of the human body and well known in herbal medicine as a restorative agent. Several studies have shown that Rg3 has a potent anti-tumor effect on various cancer cell lines. However, Rg3-induced apoptosis in B16F10 melanoma cancer cells is not well understood. In the present study, we tested whether ginsenoside Rg3 could induce apoptosis in B16F10 melanoma cells. We found that Rg3 could inhibit B16F10 melanoma cell viability in a dose-dependent manner, but not normal cells, such as EA.hy.926 and NIH3T3 cells. We also found that Rg3 could induce apoptosis in B16F10 melanoma cells using tunnel-staining assay in a dose-dependent manner. Rg3 treatment induces the phosphorylation of p38 and the expression of Bax, but it inhibits the expressions of the phosphorylation of focal adhesion kinase Bcl2 and pro-caspase3. Taken together, our data suggest that Rg3 could be useful as an anti-cancer agent in B16F10 melanoma cells.

Effects of Antimicrobial and Cytotoxicity of Undaria pinnatifida Sporophyll Fractions (미역귀 분획물의 항균 · 암세포 성장저지 효과)

  • Park, Soung-Young;Jung, Young-Hwa;Shin, Mi-Ok;Jung, Bok-Mi;Bae, Song-Ja
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.6
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    • pp.765-770
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    • 2005
  • In this study, we investigated antimicrobial and cytotoxicity effects of Undaria pinnatifida Sporophyll, which using methanol, dichloromethane and ethanol were extracted and fractionated into four different types: methanol (UPMM), hexane (UPMH), butanol (UPMB) and aqueous (UPMA). The antimicrobial activity was increased in proportion to its concentration by the paper disc method. Among the solvent fractions, UPMM and UPMB showed relatively strong antimicrobial activities in the order. Among various partition layers, the methanol partition layer (UPMM) was showed the strongest cytotoxic effects on all cancer cell lines. We also observed quinone reductase (QR) induced effects in all fraction layers of UP on HepG2 cells. The QR induced effects of UPMH on HepG2 cells at $320\mu g/mL$ concentration indicated 2.36 with a control value of 1.0.

Regulatory Mechanism in Tissue-specific Expression of Insulin-like Growth Factor-I Gene (Insulin-like growth factor-I 유전자의 조직 특이적 발현에 대한 조절기전)

  • 안미라
    • KSBB Journal
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    • v.18 no.4
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    • pp.329-334
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    • 2003
  • The present study was aimed at investigating the regulatory mechanism in tissue-specific expression of insulin-like growth factor-I (IGF-I) gene. The expression of IGF-I gene was determined by a solution hybridization/RNase protection assay using total RNA prepared from rat liver or brain of various ages. The levels of IGF-I transcripts were increased in liver gradually after birth, but decreased in brain. By using an oligonucleotide (FRE) corresponding to the C/EBP binding site of the rat IGF-I exon 1, multiple forms of C/EBP${\alpha}$ and C/EBP${\beta}$ proteins, which have DNA-binding activity, were detected in the rat liver or brain. Western immunoblot and southwestern analyses show that p42$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\alpha}$/, p35$\^$C/EBP${\alpha}$/, p38$\^$C/EBP${\beta}$/, and p35$\^$C/EBP${\beta}$ form specific complexes with the IGF-I exon 1 oligonucleotide in liver nuclear extract and that p42$\^$C/EBP${\alpha}$/ and p38$\^$C/EBP${\beta}$/ form complexes in brain. These data suggest that the formation of FRE-C/EBP isoform complexes may play important roles in the tissue-specific regulation of IGF-I gene expression.

Effects of Cultured Oyster Powder on Food Quality of Soybean Pastes (양식산 굴 첨가가 된장의 품질에 미치는 영향)

  • Kim, Jin-Soo;Heu, Min-Soo
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.208-215
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    • 2004
  • As a part of a study on the effective use of cultured oyster, oyster powder-added soybean pastes were prepared and its characteristics were examined. As the concentration of oyster powder added to commercial soybean paste was increased, the moisture level decreased, the crude and amino nitrogen content increased with no change in the salinity and titratable acidities. Also, the color of the soybean paste changed to a darker color as the concentration of oyster powder added to commercial soybean paste increased. Judging from the results of the above experiment and sensory evaluation, the optimal additive concentration of oyster powder to commercial soybean paste was revealed as 9%. The total amino acid content of oyster powder-added soybean paste was 2% higher than that of commercial soybean paste. The ratio of calcium and phosphorus of oyster powder-added soybean paste was about 1:2-2:1, an improvement in calcium absorption as compared to commercial soybean paste. Functional fatty acids, such as 20:5n-3 and 22:6n-3, were not detected in commercial soybean paste but detected in oyster powder-added soybean paste. The major free amino acids in oyster powder-added soybean paste were glutamic acid and aspartic acids. According to the above results, the nutritional, functional and taste properties of commercial soybean paste were improved while reducing preparation time of the housewife.