• Title/Summary/Keyword: $\beta$-glucuronidase activity

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Biotransformation of Glycyrrhizin by Human Intestinal Bacteria and its Relation to Biological Activities

  • Kim, Dong-Hyun;Hong, Sung-Woon;Kim, Byung-Taek;Bae, Eun-Ah;Park, Hae-Young;Han, Myung-Joo
    • Archives of Pharmacal Research
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    • v.23 no.2
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    • pp.172-173
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    • 2000
  • The relationship between the metabolites of glycyrrhizin (18$\beta$-glycyrrhetinic acid-3-O--D-glu-curonopyranosyl-($1{\rightarrow}2$)-$\beta$-D-glucuronide, CL) and their biological activities was investigated. By human intestinal microflora, CL was metabolized to 18$\beta$-glycyrrhetinic acid (GA) as a main product and to 18$\beta$-glycyrrhetinic acid-3-O-$\beta$-D-glucuronide (GAMG) as a minor product. The former reaction was catalyzed by Eubacterium L-8 and the latter was by Streptococcus LJ-22. Among GL and its metabolites, GA and GAMG had more potent in vitro anti-platelet aggregation activity than GL. GA also showed the most potent cytotoxicity against tumor cell lines and the potent inhibitory activity on rotavirus infection as well as growth of Helicobacter pylori. GAMG, the minor metabolite of GL, was the sweetest.

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Biotransformation of flavonoid-7-O-glucuronides by $\beta$-glucuronidases

  • Choi, Ran-Joo;Ha, In-Jin;Choi, Jae-Sue;Park, You-Mie;Kim, Yeong-Shik
    • Natural Product Sciences
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    • v.16 no.1
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    • pp.1-5
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    • 2010
  • $\beta$-Glucuronidases (E.C. 3.2.1.31) from Escherichia coli, Helix pomatia, and bovine liver activity have been investigated on 7-O-glucuronides (baicalin, wogonoside, and luteolin-7-O-glucuronide) and 3-O-glucuronides (quercetin-3-O-glucuronide and kaempferol-3-O-glucuronide). Bovine liver enzyme was not active on any of these substrates. E. coli and H. pomatia enzymes were active on 7-O-glucuronides, however, 3-O-glucuronides were resistant to $\beta$-glucuronidase hydrolysis. These results suggest that glucuronic acid at 7-position is more susceptible to E. coli and H. pomatia $\beta$-glucuronidases than that at 3-position. In addition, the subtle difference of aglycone structure on 7-O-glucuronides affected the preference of enzyme. E. coli enzyme was favorable for the hydrolysis of baicalin, however, H. pomatia enzyme was found to be efficient for the hydrolysis of wogonoside. Both enzymes showed the similar hydrolytic activity towards luteolin-7-O-glucuronide. When the Scutellaria baicalensis crude extract was subjected to enzymatic hydrolysis, baicalin and wogonoside were successfully converted to their aglycone counterparts with H. pomatia at 50 mM sodium bicarbonate buffer pH 4.0. Accordingly, the enzymatic transformation of glycosides may be quite useful in preparing aglycones under mild conditions.

Isolation and Properties of $\beta$-N-Acetyl-D-glucosaminidase B from Rat Uterus

  • Jung, Jin-Ha;Yang, Chul-Hak
    • Bulletin of the Korean Chemical Society
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    • v.4 no.3
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    • pp.139-143
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    • 1983
  • ${\beta}$-N-Acetyl-D-glucosaminidase B was highly purified with the following sequence of steps; DEAE-cellulose, CM-cellulose, and Sephadex G-200 gel filtration chromatograpies. The specific activity of the purified ${\beta}$ -N-acetyl-D-glucosaminidase B was 2.2 units/mg protein with 12.9 % yield and 196.2 fold purity. The purified ${\beta}$-N-acetyl-D-glucosaminidase B showed single band on polyacrylamide gel electrophoresis. The final preparation of ${\beta}$ -N-acetyl-D-glucosaminidase B was completely free friom arylsulfatase and ${\beta}$-glucuronidase. ${\beta}$ -N-Acetyl-D-glucosaminidase B had pH optimum of 4.5 in 0.5 M sodium citrate buffer. The molecular weight of ${\beta}$-N-acetyl-D-glucosaminidase B was 133,000 by Sephadex G-200 gel filtration. The Km value of ${\beta}$-N-acetyl-D-glucosaminidase B using p-nitrophenyl-N-acetyl-${\beta}$-D-glucosaminide as substrate was 1.0 mM and $V_{max}$ was 0.014 ${\mu}$ mole/min. ${\beta}$-N-Acetyl-D-glucosaminidase B was stable at $55^{circ}C$ for 70 minutes. The crude ${\beta}$ -N-acetyl-D-glucosamiinidase in 70 % ammonium sulfate retained 93 % activity after 7 months storage at -$55^{circ}C$. Bovine serum albumin, sodium chloride, and phosphate activated ${\beta}$ -N-Acetyl-D-glucosaminidase B. N-Acetyl-D-glucosamine, ${\alpha}$-methyl-D-mannoside, and acetate inhibited ${\beta}$ -N-acetyl-D-glucosaminidase B.

Cytochemical Characteristics of Blood and Bone Marrow Cells in Dog (개의 혈액 및 골수세포의 세포화학적 특성)

  • Lee Chang Woo;Hasegawa A.;Ono K.;Goitsuka R.;Yang M.P.
    • Journal of Veterinary Clinics
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    • v.7 no.1
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    • pp.429-438
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    • 1990
  • The cytochemical characteristics of the hematopoietic cells in blood and bone marrow from 3 clinically healthy dogs were examined using a battery of cytochemical stains. Alkaline phosphotase activity was demonstrated in eosinophilic series and occassionally in progranulocytes. A variety of cells exhibited acid phosphatase activity, but tartrate-resistant acid phosphatase activity was seen only in eosinophilic series. Peroxidase activity was observed in myeloblasts to segmented cells of granulocytic series and in monocytes. ${\alpha}$-naphthyl acetate esterase activity was found in monocytes and occassionally in lymphocytes. Naphthyl-AS-D-chloroacetate esterase marked neutrophilic series from myeloblasts to segmented cells. ${\beta}$-Glucuronidase activity was detected in a variety of cells except the cells of erythrocytic series. Periodic acid Schiff stain-positive granules were demonstrated in the neutrophilic and eosinophilic series from myelocytes to segmented cells and in monocytes and occassionally in lymphocytes. Sudan black B stain-positive granules marked granulocytic series from myeloblasts to segmented tells and monocytes.

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The effect of Sagunja-tang on TNBS-induced Inflammatory Bowel Disease in Mouse (사군자탕(四君子湯)이 TNBS로 유발(誘發)된 생쥐의 염증성(炎症性) 장질환(腸疾患)에 미치는 영향(影響))

  • Hong, Sang-Sun;Ryu, Bong-Ha;Yoon, Seong-Woo;Kim, Jin-Sung
    • The Journal of Internal Korean Medicine
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    • v.31 no.4
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    • pp.731-751
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    • 2010
  • Objectives : The present study aimed to find out the effect of Sagunja-tang on the prevention and treatment of inflammatory bowel disease using mice with TNBS-induced inflammatory bowel disease. Methods : Mice with TNBS-induced inflammatory bowel disease were medicated with Sagunja-tang, and the weight changes, colon length, lipid peroxidation, and myeloperoxidase activity were observed. Levels of the inflammatory markers interleukin (IL)-$1{\beta}$ and cyclooxygenase-2 (COX-2), its transcription factor activation, phospho-NF-${\kappa}$B (pp65), in the colon by enzyme-linked immunosorbent assay and immunoblot analysis were also measured. Finally, the activation of fecal bacterial enzyme, ${\beta}$-glucuronidase and degradation activation of fecal glycosaminoglycan (GAG) and hyaluronic acid were observed. Results : We found that oral administration of Sagunja-tang inhibited TNBS-induced colon shortening and also inhibited myeloperoxidase activity in the colon of mice as well as IL-$1{\beta}$ and COX-2 expression. Sagunja-tang also inhibited TNBSinduced lipid peroxidation and pp65 activation in the colon of mice. In addition, Sagunja-tang inhibited ${\beta}$-glucuronidase activation and fecal hyaluronic acid degradation activation. Conclusions : It is supposed that Sagunja-tang has a potential therapeutic effect on inflammatory bowel disease through the inhibition of both NF-${\kappa}$B activation and lipid peroxidation, and the improvement of intestinal conditions.

Fermentation of Antler and its Biological Activity (녹용발효와 생리활성)

  • Kim, Dong-Hyun;Han, Sang-Bum;Park, Ju-Suk;Han, Myung-Joo
    • Korean Journal of Pharmacognosy
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    • v.25 no.3
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    • pp.233-237
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    • 1994
  • Bacillus P-92 which fermented antler was isolated from intestinal bacteria. The biological activites, carbon clearance and growth activity of lactic acid bacteria, of fermented antler was better than those of untreated antler. The enyzmes activities, ${\beta}-glucosidase,\;{\beta}-glucuronidase$ and tryptophanase, of intestinal bacteria of mice treated with fermented antler were lower than those of mice treated with untreated antler, although those of mice treated with fermented antler or untreated antler were higher than those of control. Biological activity of the antler seems to be increased by fermentation.

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Perfusion Cultivation of Transgenic Nicotiana tabacum Suspensions in Bioreactor for Recombinant Protein Production

  • Lee Sang-Yoon;Kim Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.5
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    • pp.673-677
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    • 2006
  • A perfusion culture of transgenic Nicotiana tabacum cell suspensions, transformed to express recombinant glucuronidase (GUS), was successfully performed in a 5-1 stirred tank bioreactor. With 0.1 $day^{-1}$ of perfusion rate, the maximum dry cell weight (DCW) reached to 29.5 g/l in 16 days, which was 2.1-fold higher than the obtained in batch culture (14.3 g/l). In terms of the production of GUS, the volumetric activity could be increased up to 12.8 U/ml by using perfusion, compared with 4.9 U/ml in batch culture. The specific GUS activities in both perfusion and batch cultures were maintained at similar levels, 200-400 U/g DCW. Consequently, a perfusion culture could be a good strategy for the enhanced production of recombinant proteins in a plant cell culture system.

Studies on the Pharmacological Actions of Cactus:Identification of Its anti-Inflammatory Effect

  • Park, Eun-Hee;Kahng, Ja-Hoon;Paek, Eun-Ah
    • Archives of Pharmacal Research
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    • v.21 no.1
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    • pp.30-34
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    • 1998
  • The ethanol extracts of Opuntia ficus-indica fructus (EEOF) and Opuntia ficus-indica stem (EEOS) were prepared and used to evaluate the pharmacological effects of cactus. Both the extracts inhibited the writhing syndrome induced by acetic acid, indicating that they contains analgesic effect. The oral administrations of EEOF and EEOS suppressed carrageenan-induced rat paw edema and also showed potent inhibition in the leukocyte migration of CMC-pouch model in rats. Moreover, the extracts suppressed the release of $\beta$-glucuronidase, a lysosomal enzyme in rat neutrophils. It was also noted that the extracts showed the protective effect on gastric mucosal layers. From the results it is suggested that the cactus extracts contain anti-inflammatory action having protective effect against gastric lesions.

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Bioconversion of Flavones During Fermentation in Milk Containing Scutellaria baicalensis Extract by Lactobacillus brevis

  • Xu, Chen;Ji, Geun-Eog
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1422-1427
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    • 2013
  • Scutellaria baicalensis (SB), a traditional herb with high pharmacological value, contains more than 10% flavone by weight. To improve the biological activity of flavones in SB, we aimed to enhance the bioconversion of baicalin (BG) to baicalein (B) and wogonoside (WG) to wogonin (W) in SB during fermentation using beta-glucuronidase produced from Lactobacillus brevis RO1. After activation, L. brevis RO1 was cultured in milk containing SB root extract with various carbon or nitrogen sources at $37^{\circ}C$ for 72 h. During fermentation, the growth patterns of L. brevis RO1 and changes in the flavone content were assessed using thin-layer chromatography and high-performance liquid chromatography. After 72 h of fermentation, the concentrations of B and W in the control group increased by only 0.15 and 0.12 mM, respectively, whereas they increased by 0.57 and 0.24 mM in the fish peptone group. The production of B and W was enhanced by the addition of 0.4% fish peptone, which not only improved the growth of L. brevis RO1 (p < 0.001) but also enhanced the bioconversion of flavones. In conclusion, the bioconversion of flavones in SB may provide a potential application for the enhancement of the functional components in SB.

Nephrotoxicity Assessment by Determination of Alanine Aminopeptidase (AAP) and $\beta$-Glucuronidase(GRS) in Rat (랫트뇨중의 Alanine Aminopeptidase와 $\beta$-Glucuronidase 측정에 의한 신장독성 평가에 관하여)

  • Shin Dong-Hwan;Lee Chang Wo
    • Journal of Veterinary Clinics
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    • v.8 no.1
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    • pp.31-45
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    • 1991
  • Present experiment was performed in order to establish the optimal reaction conditions for determination of urinary AAP and GRS activities and to investigate the applicability of urinary AAP and GRS in nephrotoxicity test in rat. The results were as follows ; 1. The optimal pH of phosphate buffer for determination of urinary AAP activity was 7.8. 2. The Michaelis constant of urinary AAP ranged from 0.8 to 1.0mmol/$\ell$ 3. The optimal wave length for determination of urinary GRS activity was 405nm. 4. The optimal pH of acetate buffer for determination of urinary GRS activity was 5.6. 5. The Michaelis constant of urinary GRS ranged from 0.65~0.79mmo1/$\ell$. 6. The AAP activities in gel-filtered samples were significantly higher than those in crude samples. Mean values of AAP activities in gel-filtered samples and crude samples were 29$\pm$20 and 20$\pm$13U/$\ell$, respectively. 7. There was not significant difference between gel-filtered samples and crude samples in urinary GRS activities. Mean values of GRS activities in gel-filtered samples and crude samples were 57$\pm$40 and 56$\pm$39U/$\ell$, respectively. 8. Limits of linearity of urinary AAP and GRS activities were 2.0 and 3.6U/$\ell$, respectively. 9. Within-run imprecisions of the assays, were acceptable, as the coefficients of the AAP activities ranged from 5.5 to 6.3% and those of GRS activities ranged from 1.4 to 6.2%, respectively. 10. Urinary AAP excretion was 675$\pm$227mu/24hrs.kg before administration of potassium dichromate, and increased significantly to 4246$\pm$2567mU/24hrs.kg within 24 hours after administration of potassium dichromate. 11. Urinary GRS excretion did not increase significantly after administration of potassim dichromate. 12. From these findings it is concluded that urinary AAP excretion is early and sensitive Indicator to detect kidney damage in nephrotoxicity experiment.

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