• Title/Summary/Keyword: Rb2

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Enzymatic Biotransformation of Ginsenoside Rb1 and Gypenoside XVII into Ginsenosides Rd and F2 by Recombinant β-glucosidase from Flavobacterium johnsoniae

  • Hong, Hao;Cui, Chang-Hao;Kim, Jin-Kwang;Jin, Feng-Xie;Kim, Sun-Chang;Im, Wan-Taek
    • Journal of Ginseng Research
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    • v.36 no.4
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    • pp.418-424
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    • 2012
  • This study focused on the enzymatic biotransformation of the major ginsenoside Rb1 into Rd for the mass production of minor ginsenosides using a novel recombinant ${\beta}$-glucosidase from Flavobacterium johnsoniae. The gene (bglF3) consisting of 2,235 bp (744 amino acid residues) was cloned and the recombinant enzyme overexpressed in Escherichia coli BL21(DE3) was characterized. This enzyme could transform ginsenoside Rb1 and gypenoside XVII to the ginsenosides Rd and F2, respectively. The glutathione S-transferase (GST) fused BglF3 was purified with GST-bind agarose resin and characterized. The kinetic parameters for ${\beta}$-glucosidase had apparent $K_m$ values of $0.91{\pm}0.02$ and $2.84{\pm}0.05$ mM and $V_{max}$ values of $5.75{\pm}0.12$ and $0.71{\pm}0.01{\mu}mol{\cdot}min^{-1}{\cdot}mg$ of $protein^{-1}$ against p-nitrophenyl-${\beta}$-D-glucopyranoside and Rb1, respectively. At optimal conditions of pH 6.0 and $37^{\circ}C$, BglF3 could only hydrolyze the outer glucose moiety of ginsenoside Rb1 and gypenoside XVII at the C-20 position of aglycon into ginsenosides Rd and F2, respectively. These results indicate that the recombinant BglF3 could be useful for the mass production of ginsenosides Rd and F2 in the pharmaceutical or cosmetic industry.

Effect of Acid Hydrolyzates of Ginseng Saponins on Lipid Metabolism in Rat Epi didymal Adipose Tissue (인삼Saponin의 산가수분해물이 Epididymal Adipose Tissue의 지방대식에 미치는 영향)

  • Do, Jae-Ho;Kim, Sang-Dal
    • Journal of Ginseng Research
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    • v.6 no.2
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    • pp.123-130
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    • 1982
  • Studies were carried out to clarify the effect of ginsenoside-Rbl -Rbr and acid hydrolyzatps of ginsenoside-Rbl, -Rb2 (HRbl, HRbf) on lipolysis and lipogenesis induced by epinephrine, glucagon, ACTH (adrenocorticotrophic hormone), TSH (thyroid-stimulating hormone) and insulin in rat adipose tissilr. HRbl , HRb2 slightly inhibited lipolysis induced by epinephine. glucagon and TSH. ACTH-induced lipolysis in fat tissue slices was significantly inhibited by ginsenoside -Rbl, -Rb2, HRbl and HRb2, particulary HRb2. None of ginsenoside-Rbl, -Rb2, HRbl and HRb2 accelerated insulin-stimulated lipogenesis in fat calls. Among ginseng products, extract powder (freeze dried), extract powder (spray dried), red ginseng powder inhibited ACTH-induced lipolysis in adipose tissue slices, but red ginseng extract not affect them.

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The Dyeing Properties of Woody Fiber Regenerated from Waste MDF by Reactive Dyes (반응성염료에 의한 폐MDF 재생 목질섬유의 염색특성)

  • Ju, Seon-Gyeong;Roh, JeongKwan
    • Journal of the Korean Wood Science and Technology
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    • v.47 no.2
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    • pp.163-177
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    • 2019
  • This study aims to review the relations between the dyeing conditions (i.e., dye concentration, addition amounts of salt and alkali, and dyeing temperature) and dyeing properties and color fastness to light for identifying the optimal dyeing conditions when dyed regenerated woody fibers were obtained through the defibration of waste medium density fiberboard (MDF) using reactive Red H-E3B (Bis-monochlorotriazine (MCT)/MCT type) and reactive Red RB133% (Bis-MCT/Vinyl sulphone type). The dyeing yield (K/S) obtained using two types of reactive dyes increased as the dye concentration increased by 1-10% (on the weight of fiber (OWF)). In addition, the K/S of H-E3B was higher than that of RB133% irrespective of the dye concentration. The color difference of H-E3B after ultraviolet (UV) radiation was lower than that of RB133%, denoting good resistance to discoloration by UV. As the amount of sodium sulfate increased, the color difference and K/S also increased, and the adequate salt content was determined to be 50-70 g/L. Further, the color difference and K/S significantly increased only the addition of 2 g/L of sodium carbonate; however, almost no difference was observed when more than 2 g/L of sodium carbonate was added. The addition amount of sodium carbonate was adequate 5-10 g/L to dyeing the fiber and the pH at this addition level was 10. The dyeing yield of H-E3B increased when the dyeing temperature increased; however, it subsequently decreased after the dyeing temperature became $80^{\circ}C$. The dyeing yield of RB133% was almost the same up to $60-70^{\circ}C$ but declined subsequently. Thus, the adequate temperatures were $80^{\circ}C$ and $60^{\circ}C$ for H-E3B and RB133%, respectively. If the waste MDF woody fiber was dyed under the aforementioned optimal conditions, dyed regenerated woody fiber can be obtained having the following colors: 1.5 to 2.0R with the H-E3B dye and 9.6 to 10.0 PR with RB133%.

Effects of Various Nucleotides on the Membrane Permeability (Nucleotides가 세포막 투과도에 미치는 영향)

  • Lee, Joong-Woo;Jeong, Seong-Woo
    • The Korean Journal of Physiology
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    • v.23 no.1
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    • pp.13-21
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    • 1989
  • The present study was designed to investigate i) the action of various nucleotides on membrane permeability of rat red blood cell and hepatocyte for $Na^{+}$ and $Rb^{+}$ ii) the characteristics of purinoceptors on these cell membranes. Blood from Sprague-Dawley rats was obtained by carotid arterial cannulation. Red blood cells were then washed 3 times with saline at $4{\circ}C$. Hepatic parenchymal cells were isolated from rat livers by using a modification of the Berry and Friend (1969) method. For the $Na^{+}$ influx studies, isolated RBC and hepatocyte were incubated in incubation medium containing $^{22}Na^{+}0.2\;{\mu}Ci/ml$ at $37^{\circ}C$. After various time intervals samples were removed from the incubation flask and washed out 3 times with ice-cold washing solutions. Cells were destroyed by adding Triton X-100 and TCA solution. After centrifugation, the supernatants were assayed for $^{22}Na^{+}$ by gamma counter. $^{86}Rb^{+}$ was used to simulate $K^{+}$ in these $K^{+}efflux$ studies. Isolated hepatocytes were incubated for 60 min in the loading solution containing $^{86}Rb^{+}\;10\;{\mu}Ci/ml$ at $37^{\circ}C$. After loading, the cells washed out 3 times by centrifugation with washing solution. The cells were incubated in buffer solution at $37^{\circ}C$. At intervals thereafter, samples were removed and centrifuged. The supernatants were analyzed for $^{86}Rb^{+}$ by liquid scintillation counter. The main results of the experiments were: 1) ATP and ATPP increased in both $^{22}Na^{+}$ influx and $^{86}Rb^{+}$ efflux in the red blood cell. Although ADP showed a tendency to increase in RBC membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$, the changes were not significantly different from the control. 2) The Significant changes in $^{22}Na^{+}$ and $^{86}Rb^{+}$ flux by ATP were also demonstrated in hepatocyte. ATPP and ADP showed a tendency to increase in hepatocyte membrane permeability for both ions. 3) Other nucleoside triphosphates-ITP, GTP and CTP-did not change in membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$ in RBC and hepatocyte. In conclusion, not only ATP but also ATPP activate purinoceptors and change in membrane permeability for $Na^{+}$ and $K^{+}$. In order to activate purinoceptors on the cell membrane, the nucleotides have to possess intact adenine moiety and three phosphates or more in its molecule.

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Effect of White Ginseng on the Proliferation of Mouse Peritoneal Macrophages and Their Nitric Oxide Synthesis (백삼성분이 마우스 복강 탐식세포의 증식 및 Nitric Oxide 생성에 미치는 영향)

  • 김주원;배지현
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.4
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    • pp.484-490
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    • 1997
  • In an attempt to investigate the effects of white ginseng on the proliferation and the nitric oxide(NO) secretion of mouse peritoneal macrophages, which are the first mai or defense phagocytes in the immune system, the studies have been carried out. In the macrophage proliferation assay using the $^3$H-thymidine incorporation, the total saponin or Ginsenoside Rb$_2$ were added to the medium at the concentration of 0 to 256$\mu\textrm{g}$/$m\ell$. DNA synthesis of the macrophage was increased at 64$\mu\textrm{g}$/$m\ell$ of total saponin and either 16$\mu\textrm{g}$/$m\ell$ or 64$\mu\textrm{g}$/$m\ell$ of Ginsenoside Rb$_2$, respectively. Also, the effect o(white ginseng on the nitric oxide secretion of the macrophages was investigated. The addition of either total saponin or Ginsenoside Rb$_2$ at the concentration of 20$\mu\textrm{g}$/$m\ell$ significantly increased the secretion of NO from the macrophages. The nitric oxide synthase (NOS) gene expression which is responsible for the synthesis of the nitric oxide has been studied using reverse transcription polymerase chain reaction. In RT-PCR, the $\beta$-actin and nos gene expression have been analyzed. 20$\mu\textrm{g}$/$m\ell$ of either total saponin or Ginsenoside Rb$_2$ increased nos gene expression of the macrophages.

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Relationship among Ginsenosides of Panax ginseng Root under the Variation of Mineral Nutrients (무기영양변화에서 인삼근Ginsenoside의 상호관계)

  • Lee, Mee-Kyoung;Min, Jin-Sook;Park, Hoon
    • Journal of Ginseng Research
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    • v.10 no.1
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    • pp.101-107
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    • 1986
  • Relationships among ginsenosides, panaxadiol(PD), panaxatriol(PT), and total saponin(TS) in Panax ginseng root (2nd Year) grown with culture solotion different in nitrogen, phosphorus and potassium level were analyzed by simple correlation, multiple regression and standard partial regression coefficient. The closeness between ginsenosides by simple correlation was closely related with the similarity of molecular structure. The content of PT was much attributed to Re and Rg1. The contribution order of ginsenosides for PD was Rb1>Rb2$\geq$Rd>Rc. There was significant positive correlation between PT and PD but PD increased more rapidly than PT. Thus total saponin depended much on PD and PT/PD decreased with the increase of total saponin content. All ginsenosides, especially Re showed decreasing tendency with the increase of root weight.

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Preparation and Evaluation of New Adsorbent (Paper Sludge) in the Treatment of Waste Water of Textile Industry (섬유산업 폐수 처리시 새로운 종이슬러지 흡착제의 제조와 평가)

  • Elaziouti, A.;Laouedj, N.;Segheir, A.A.
    • Journal of the Korean Chemical Society
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    • v.54 no.1
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    • pp.115-119
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    • 2010
  • Neutral red5 (BR 5) adsorption from aqueous solution was investigated. New solid adsorbent was prepared from paper sludge incinerated at $250^{\circ}C$ during 2 hours. Experiments were carried out, in batch system, to remove RB5 dye from the synthetic waste water. Maximum adsorption of 374.98 mg/g at pH 5, at room temperature and contact time of 80 min was obtained. The Langmuir and Freundlich adsorption models were applied to interpret the equilibrium isotherm. The Freundlich isotherm fits quite well with the experimental data (with highly regression coefficient $R^2$). The incinerated sludge solid samples untreated and treated with BR5 dye were characterized by FT-IR spectrophotometer.

Postprandial hypoglycemic effects of mulberry twig and root bark in vivo and in vitro (In vivo와 in vitro에서 상지 및 상백피 에탄올추출물이 식후 혈당 상승 억제 조절에 미치는 영향)

  • Park, Soo Yeon;Jin, Bo Ra;Lee, Yu Rim;Kim, You Jin;Park, Jeong Bin;Jeon, Young Hee;Choi, Sang Won;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.49 no.1
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    • pp.18-27
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    • 2016
  • Purpose: Our previous study demonstrated the hypoglycemic effects of mulberry (Morus alba L.) leaf and the underlying mechanisms. Here we explored the potency of mulberry twigs (TW) and root barks (RB) in postprandial hypoglycemic effects in vitro and in vivo. Methods: The major components of TW and RB were determined by high performance liquid chromatography (HPLC). Alpha-glucosidase inhibition and glucose/fructose uptake inhibition in Caco-2 cells were determined for TW, RB, and their major components, followed by an oral sugar tolerance test (OSTT) in streptozotocin-induced diabetic rats. Male Wistar rats were fed a high-fat diet for 2 weeks and then a single dose of streptozotocin (35 mg/kg B.W) was administered by intraperitoneal injection. Rats with fasting blood glucose levels above 126 mg/dL were randomly divided into 5 groups (n = 8/group) for the following treatments by gavage for 4 weeks: vehicle (normal control and diabetic control), 200 mg/kg B.W of TW or RB or 100 mg/kg B.W of oxyresveratrol (OXY). Results: OXY and mulberroside A were identified as the major components of TW and OXY, mongolicin, and kuwanon H for RB. A significant inhibitory activity on ${\alpha}-glucosidase$ was found for TW, RB, and OXY (p = 0.0099). There was a dose-dependent inhibition of TW and RB on the intestinal sugar uptakes in Caco-2 cells, showing a greater impact on fructose compared to glucose. The OSTT showed that TW and RB significantly delayed time to maximal concentration (p = 0.0088) and decreased maximal concentration (p = 0.0043) compared to the control group. Conclusion: These results suggest that TW and RB may have a postprandial hypoglycemic effect, particularly in the case of high fructose or sucrose intake. OXY was suggested as a contributor to the hypoglycemic effect of TW and RB. Further studies are needed for the systemic effect of TW and RB in circulation.

Development of a Screening System for Drugs Against Human Papillomavirus-Associated Cervical Cancer: Based On E7-Rb Binding

  • Cho, Young-Sik;Cho, Cheong-Weon;Kang, Jeong-Woo;Cho, Min-Chul;Lee, Kyung-Ae;Shim, Jung-Hyun;Kwon, Our-Han;Choe, Yong-Kyung;Park, Sue-Nie;Yoon, Do-Young
    • BMB Reports
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    • v.34 no.1
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    • pp.80-84
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    • 2001
  • The human papillomavirus E7 protein can form a specific complex with a retinoblastoma tumor suppressor gene product (p105-Rb) that results in the release of the E2F transcription factor, which is critical for the growth-deregulation and transforming properties of the viral E7 oncoprotein. In an attempt to apply interaction between the E7 oncoprotein and a target cellular protein Rb for an in vitro screening system for drugs against human papillomavirus infection, we primarily investigated the E7Rb binding through a pull down assay and enzyme-linked immunosorbent assay. The pull down assay showed that both glutathione S-transferase-tagged E7 and His-tagged E7 immobilized on resins specifically produced complexes with bacterially expressed Rb in a dose-dependent manner, as determined by immunoblot analyses. This result coincided with that of an enzyme-linked immunosorbent assay, which is a useful system for the mass screening of potential drugs. Taken together, this screening system (based on the interaction between E7 and Rb) can be a promising system in the development of drugs against cervical cancers caused by human papillomavirus infection.

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