Relevant bibliographies by topics / Β-Glucuronidases

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  2. Dissertations / Theses
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Academic literature on the topic 'Β-Glucuronidases'

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Published: 19 February 2023

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Journal articles on the topic "Β-Glucuronidases"

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Bae, Hyung-Sup, Young-Suk Kim, Ki-Ho Cho, Kyung-Sup Lee, Jung-Jin Kim, Hae-Ung Lee, and Dong-Hyun Kim. "Hepatoprotective Activity of Reduohanxiao-tang (Yuldahanso-tang) is Related to the Inhibition of β-Glucuronidase." American Journal of Chinese Medicine 31, no. 01 (January 2003): 111–17. http://dx.doi.org/10.1142/s0192415x03000722.

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β-Glucuronidase-inhibitory and hepatoprotective effects of Reduohanxiao-tang (Yuldahanso-tang), which has been used for liver diseases and stroke, on carbon tetrachloride (CCl4)-induced hepatotoxicity of rats were investigated. Reduohanxiao-tang potently inhibited β-glucuronidases. Serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactic acid dehydrogenase (LDH) levels of the CCl4 group orally treated with Reduohanxiao-tang (100 mg/kg) were lowered to 54%, 71.5% and 66.1% of the CCl4-treated control group, respectively. Among the ingredients of the Reduohanxiao-tang, the rhizomes of Pueraria thunbergiana and it Scutellaria baicalensis potently inhibited β-glucuronidases and protected against CCl4-induced liver injury. Orally administered puerarin, which is a main component of Pueraria thunbergiana, showed potent hepatoprotective activity, but did not inhibit β-glucuronidase. However, daidzein, which is produced from puerarin by human intestinal bacteria, potently inhibited β-glucuronidase. These results suggest that β-glucuronidase inhibition by herbal medicines may protect a gainst CCl4-induced liver injury.
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Bai, Yue, Lu Chen, Yun-Feng Cao, Xu-Dong Hou, Shou-Ning Jia, Qi Zhou, Yu-Qi He, and Jie Hou. "Beta-Glucuronidase Inhibition by Constituents of Mulberry Bark." Planta Medica 87, no. 08 (March 17, 2021): 631–41. http://dx.doi.org/10.1055/a-1402-6431.

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AbstractIntestinal bacterial β-glucuronidases, the key enzymes responsible for the hydrolysis of various glucuronides into free aglycone, have been recognized as key targets for treating various intestinal diseases. This study aimed to investigate the inhibitory effects and mechanisms of the Mulberry bark constituents on E. coli β-glucuronidase (EcGUS), the most abundant β-glucuronidases produced by intestinal bacteria. The results showed that the flavonoids isolated from Mulberry bark could strongly inhibit E. coli β-glucuronidase, with IC50 values ranging from 1.12 µM to 10.63 µM, which were more potent than D-glucaric acid-1,4-lactone. Furthermore, the mode of inhibition of 5 flavonoids with strong E. coli β-glucuronidase inhibitory activity (IC50 ≤ 5 µM) was carefully investigated by a set of kinetic assays and in silico analyses. The results demonstrated that these flavonoids were noncompetitive inhibitors against E. coli β-glucuronidase-catalyzed 4-nitrophenyl β-D-glucuronide hydrolysis, with Ki values of 0.97 µM, 2.71 µM, 3.74 µM, 3.35 µM, and 4.03 µM for morin (1), sanggenon C (2), kuwanon G (3), sanggenol A (4), and kuwanon C (5), respectively. Additionally, molecular docking simulations showed that all identified flavonoid-type E. coli β-glucuronidase inhibitors could be well-docked into E. coli β-glucuronidase at nonsubstrate binding sites, which were highly consistent with these agentsʼ noncompetitive inhibition mode. Collectively, our findings demonstrated that the flavonoids in Mulberry bark displayed strong E. coli β-glucuronidase inhibition activity, suggesting that Mulberry bark might be a promising dietary supplement for ameliorating β-glucuronidase-mediated intestinal toxicity.
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Elmassry, Moamen M., Sunghwan Kim, and Ben Busby. "Predicting drug-metagenome interactions: Variation in the microbial β-glucuronidase level in the human gut metagenomes." PLOS ONE 16, no. 1 (January 7, 2021): e0244876. http://dx.doi.org/10.1371/journal.pone.0244876.

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Characterizing the gut microbiota in terms of their capacity to interfere with drug metabolism is necessary to achieve drug efficacy and safety. Although examples of drug-microbiome interactions are well-documented, little has been reported about a computational pipeline for systematically identifying and characterizing bacterial enzymes that process particular classes of drugs. The goal of our study is to develop a computational approach that compiles drugs whose metabolism may be influenced by a particular class of microbial enzymes and that quantifies the variability in the collective level of those enzymes among individuals. The present paper describes this approach, with microbial β-glucuronidases as an example, which break down drug-glucuronide conjugates and reactivate the drugs or their metabolites. We identified 100 medications that may be metabolized by β-glucuronidases from the gut microbiome. These medications included morphine, estrogen, ibuprofen, midazolam, and their structural analogues. The analysis of metagenomic data available through the Sequence Read Archive (SRA) showed that the level of β-glucuronidase in the gut metagenomes was higher in males than in females, which provides a potential explanation for the sex-based differences in efficacy and toxicity for several drugs, reported in previous studies. Our analysis also showed that infant gut metagenomes at birth and 12 months of age have higher levels of β-glucuronidase than the metagenomes of their mothers and the implication of this observed variability was discussed in the context of breastfeeding as well as infant hyperbilirubinemia. Overall, despite important limitations discussed in this paper, our analysis provided useful insights on the role of the human gut metagenome in the variability in drug response among individuals. Importantly, this approach exploits drug and metagenome data available in public databases as well as open-source cheminformatics and bioinformatics tools to predict drug-metagenome interactions.
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Yang, Wei, Bin Wei, and Ru Yan. "Amoxapine Demonstrates Incomplete Inhibition of β-Glucuronidase Activity from Human Gut Microbiota." SLAS DISCOVERY: Advancing the Science of Drug Discovery 23, no. 1 (August 15, 2017): 76–83. http://dx.doi.org/10.1177/2472555217725264.

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Amoxapine has been demonstrated to be a potent inhibitor of Escherichia coli β-glucuronidase. This study aims to explore the factors causing unsatisfactory efficacy of amoxapine in alleviating CPT-11–induced gastrointestinal toxicity in mice and to predict the outcomes in humans. Amoxapine (100 µM) exhibited poor and varied inhibition on β-glucuronidase activity in gut microbiota from 10 healthy individuals and their pool (pool, 11.9%; individuals, 3.6%−54.4%) with IC50 >100 µM and potent inhibition toward E. coli β-glucuronidase (IC50 = 0.34 µM). p-Nitrophenol formation from p-nitrophenyl-β-D-glucuronide by pooled and individual gut microbiota fitted classical Michaelis-Menten kinetics, showing similar affinity (Km = 113–189 µM) but varied catalytic capability (Vmax = 53–556 nmol/h/mg). Interestingly, amoxapine showed distinct inhibitory effects (8.7%–100%) toward β-glucuronidases of 13 bacterial isolates (including four Enterococcus, three Streptococcus, two Escherichia, and two Staphylococcus strains; gus genes belonging to OTU1, 2 or 21) regardless of their genetic similarity or bacterial origin. In addition, amoxapine inhibited the growth of pooled and individual gut microbiota at a high concentration (6.3%–30.8%, 200 µM). Taken together, these findings partly explain the unsatisfactory efficacy of amoxapine in alleviating CPT-11–induced toxicity and predict a poor outcome of β-glucuronidase inhibition in humans, highlighting the necessity of using a human gut microbiota community for drug screening.
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Wang, Xiaoyan, Yanli Liu, Chao Wang, Xudong Feng, and Chun Li. "Properties and structures of β-glucuronidases with different transformation types of glycyrrhizin." RSC Advances 5, no. 84 (2015): 68345–50. http://dx.doi.org/10.1039/c5ra11484e.

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Correa, Sonali, Magdalena Ripoll, Erienne Jackson, Valeria Grazú, and Lorena Betancor. "Stabilization of b-Glucuronidase by Immobilization in Magnetic-Silica Hybrid Supports." Catalysts 10, no. 6 (June 13, 2020): 669. http://dx.doi.org/10.3390/catal10060669.

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β-Glucuronidases are a class of enzymes that catalyze the breakdown of complex carbohydrates. They have well documented biocatalytic applications in synthesis, therapeutics, and analytics that could benefit from enzyme immobilization and stabilization. In this work, we have explored a number of immobilization strategies for Patella vulgata β-Glucuronidase that comprised a tailored combination of biomimetic silica (Si) and magnetic nanoparticles (MNPs). The individual effect of each material on the enzyme upon immobilization was first tested. Three different immobilization strategies for covalent attachment on MNPs and different three catalysts for the deposition of Si particles were tested. We produced nine different immobilized preparations and only two of them presented negligible activity. All the preparations were in the micro-sized range (from 1299 ± 52 nm to 2101 ± 67 nm of hydrodynamic diameter). Their values for polydispersity index varied around 0.3, indicating homogeneous populations of particles with low probability of agglomeration. Storage, thermal, and operational stability were superior for the enzyme immobilized in the composite material. At 80 °C different preparations with Si and MNPs retained 40% of their initial activity after 6 h of incubation whereas the soluble enzyme lost 90% of its initial activity within 11 min. Integration of MNPs provided the advantage of reusing the biocatalyst via magnetic separation up to six times with residual activity. The hybrid material produced herein demonstrated its versatility and robustness as a support for β-Glucuronidases immobilization.
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Wong, Alexander W., Shouming He, and Stephen G. Withers. "Synthesis of 5-fluoro-β-D-glucopyranosyluronic acid fluoride and its evaluation as a mechanistic probe of Escherichia coli β-glucuronidase." Canadian Journal of Chemistry 79, no. 5-6 (May 1, 2001): 510–18. http://dx.doi.org/10.1139/v00-155.

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Synthesis of the potential mechanism-based inactivator of β-D-glucuronidases (5-fluoro-β-D-glucopyranosyluronic acid fluoride) was accomplished via a six-step process from D-glucuronic acid that involved radical bromination at C-5 and displacement of the bromide by fluoride. A key step in this process was the masking of the carboxylic acid as a phenacyl ester. This group is uniquely stable to conditions of photobromination and fluoride displacement, yet removable under very mild conditions. Incubation of the Escherichia coli β-glucuronidase with 5-fluoro-β-D-glucopyranosyluronic acid fluoride resulted in time-dependent inactivation of the enzyme through the accumulation of a covalent 5-fluoro-α-D-glucopyranosyluronic acid-enzyme. Peptic digestion of the 5-fluoro-α-D-glucopyranosyluronic acid-enzyme intermediate and subsequent analysis by liquid chromatography coupled to an electrospray ionization triple quadrupole mass spectrometer indicated the presence of a 5-fluoro-α-D-glucopyranosyluronic acid-modified peptide. This peptide was partially purified by HPLC and its sequence determined by tandem mass spectrometry in the daughter ion scan mode, permitting the identification of Glu504 as the catalytic nucleophile within the sequence ITEYGVD. This new reagent is therefore useful for the specific, mechanism-based inactivation of glycuronidases and has good potential in other studies of enzymes of this general class.Key words: β-glucuronidase, catalytic nucleophile, 5-fluoro-β-D-glucopyranosyluronic acid fluoride, electrospray MS.
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Wu, Liang, Jianbing Jiang, Yi Jin, Wouter W. Kallemeijn, Chi-Lin Kuo, Marta Artola, Wei Dai, et al. "Activity-based probes for functional interrogation of retaining β-glucuronidases." Nature Chemical Biology 13, no. 8 (June 5, 2017): 867–73. http://dx.doi.org/10.1038/nchembio.2395.

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Pollet, Rebecca M., Emma H. D'Agostino, William G. Walton, Yongmei Xu, Michael S. Little, Kristen A. Biernat, Samuel J. Pellock, et al. "An Atlas of β-Glucuronidases in the Human Intestinal Microbiome." Structure 25, no. 7 (July 2017): 967–77. http://dx.doi.org/10.1016/j.str.2017.05.003.

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Canales, I., A. Manjōn, and J. L. Iborra. "Immobilization of β-glucuronidases on an epoxy-activated polyacrylic matrix." Biotechnology Techniques 4, no. 3 (May 1990): 205–10. http://dx.doi.org/10.1007/bf00222507.

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Dissertations / Theses on the topic "Β-Glucuronidases"

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Tredwell, Gregory David. "An Investigation into β-Glucuronidases." Thesis, Griffith University, 2009. http://hdl.handle.net/10072/365204.

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Abstract: Conjugated carbohydrates, such as proteoglycans, play pivotal roles in a number of diverse biological processes. Their structural complexity encodes information that regulates intercellular communication, recognition events and cellular activity. As such, the biosynthesis and catabolism of these complex molecules requires the tight regulation of a large number of enzymes. Two enzymes that are of particular interest in the metabolism of glycosaminoglycans are exo-β-glucuronidase and the endo-β-glucuronidase, heparanase. These two glycosidases play an important role in the catabolism of glycosaminoglycan chains from extracellular matrix and basement membrane proteoglycans. Over-expression of these enzymes has been reported in a number of tumours, and they have been implicated in tumour metastasis and angiogenesis. Thus, there is considerable interest in the characterisation of these enzymes for the development of novel enzyme inhibitors. The work described in this thesis is divided into two parts. The first part describes the cloning, expression and characterisation of a novel heparanase from the pathogenic bacterium Burkholderia pseudomallei. In addition to the enzyme’s hydrolase activity against heparan sulfate, the B. pseudomallei heparanase was also found to have significant exo-β-glucuronidase activity. This is the first report of a bacterial enzyme that is capable of cleaving heparan sulfate via a hydrolase mechanism. In the second part of the work, the X-ray crystal structure of human β-glucuronidase was used to design novel enzyme inhibitors. A number of computational tools such as GRID, molecular docking and de novo ligand design were employed to generate a library of substituents that could be introduced at the C-1, C-2 and/or C-3 positions of specific carbohydrate templates that would potentially improve ligand binding to the enzyme. A number of 1-thioglucuronides, with substituted benzylic or aliphatic aglycon moieties, identified by the structure-based ligand design process, were chosen for synthesis. The central glycosidation step used selective diethylamine-mediated de-S-acetylation of methyl 2,3,4,5-tetra-O-acetyl-1-S-acetyl-1-thio-β-D-glucuronate, and subsequent reaction of the generated thiolate with an activated acceptor molecule. In the case of the substituted benzyl halide acceptors, glycosidations gave solely the β-glycosides, whereas glycosidation with substituted propyl halide acceptors was found to produce α/β-anomeric mixtures. The inhibitory activity of the prepared thioglucuronides was evaluated in vitro using a fluorogenic enzyme assay, with the most potent inhibitor showing activity down to 10-5 M against the test enzyme, bovine β-glucuronidase.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Griffith University. Institute for Glycomics.
Griffith Health
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Wei, Bin. "The distribution and function of β-glucuronidases in human gut microbiota and β-glucuronidase targeted drug discovery." Thesis, University of Macau, 2018. http://umaclib3.umac.mo/record=b3952498.

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Bohlmann, Lisa Angela. "An Investigation Into a Microbial β-Glucuronidase." Thesis, Griffith University, 2015. http://hdl.handle.net/10072/365554.

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Glycosaminoglycans (GAGs) are the most prevalent hetero-polysaccharides in the body. They are usually attached to a protein core, forming proteoglycans (PGs). PGs play crucial roles in a wide range of biological processes including cell growth, migration and differentiation. Heparan sulfate proteoglycans (HSPGs) are found in the basement membrane and the extracellular matrix (ECM), which surrounds cells and is formed by a network of fibrous proteins, glycoproteins and proteoglycans. In mammalian cells heparan sulfate (HS) is cleaved by a single β-glucuronidase: heparanase. Cleavage of HS-chains changes the integrity of the ECM and releases a variety of bioactive molecules such as growth factors, cytokines and enzymes. Heparanase is implicated in cancer and inflammation and inhibition of heparanase has been shown to reduce tumour metastasis and angiogenesis. Heparanase is therefore regarded as a promising target for anti-cancer and anti-inflammatory drug design. As the three dimensional structure of human heparanase is still not available, the progress in specific inhibitor development has been impeded.
Thesis (PhD Doctorate)
Doctor of Philosophy (PhD)
Institute for Glycomics
Science, Environment, Engineering and Technology
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Toyama, Osamu. "Klotho is a novel β-glucuronidase capable of hydrolyzing steroid β-glucuronides." Kyoto University, 2004. http://hdl.handle.net/2433/147516.

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Kodawara, Takaaki. "Inhibitory effect of ciprofloxacin on β-glucuronidase-mediated deconjugation of mycophenolic acid glucuronide." Kyoto University, 2015. http://hdl.handle.net/2433/198935.

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Ross, Kristin Coby. "Separation of Recombinant β-Glucuronidase from Transgenic Tobacco by Aqueous Two-Phase Extraction." Thesis, Virginia Tech, 2008. http://hdl.handle.net/10919/43471.

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Biopharmaceutical manufacturing is a rigorous and expensive process. Due to the medicinal nature of the product, a high purity level is required and several expensive purification steps must be utilized. Cost-effective production and purification is essential for any biopharmaceutical product to be successful and development of the fastest, most economical, and highest-yielding purification scheme is a constant engineering challenge. Commercial-scale purification schemes currently revolve around the use of multiple chromatography steps for the purification of biopharmaceutical products. Chromatography has many shortcomings including high cost, limited throughput, and complex scale up. The goal of this research was to develop an alternative, non-chromatography purification step for the separation of an acidic model protein, recombinant β-glucuronidase (rGUS), from transgenic tobacco with high yield and purity. Aqueous two-phase extraction (ATPE) is a powerful technique for separation and purification of proteins, and has the potential to replace an expensive chromatography step for the initial purification of recombinant proteins. ATPE enables high levels of target protein recovery and concentration while removing large amounts of impurities from the initial extract. Fractional factorial designs and response surface methodology were used to determine an optimized aqueous two-phase system for the purification of rGUS from transgenic tobacco. In a 13.4 % (w/w) PEG/18% (w/w) potassium phosphate system, 74% of the rGUS was recovered in the top PEG-rich phase while 90% of the native tobacco proteins were removed in the interphase and the bottom phase. A purification factor of about 20 was achieved in this process.
Master of Science
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Bundschuh, Diana Stephanie [Verfasser]. "Plazentare Expression der Endo-β-d-Glucuronidase Heparanase bei Termingeburten mit physiologischen und pathologischen Schwangerschaftsverläufen / Diana Stephanie Bundschuh." Lübeck : Zentrale Hochschulbibliothek Lübeck, 2015. http://d-nb.info/1080058036/34.

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Chapdelaine, Alexandra. "Le Bisphénol A dans la prééclampsie." Mémoire, Université de Sherbrooke, 2016. http://hdl.handle.net/11143/9606.

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Résumé : La prééclampsie (PE) est un désordre de la grossesse caractérisée par une dysfonction endothéliale faisant en sorte que l’endothélium devient moins sensible aux signaux de vasodilatation. La réponse provoquée par la liaison de la sérotonine au sous-type de récepteur S[indice inférieur 2] entraîne la libération de molécules aux propriétés vasoconstrictrices, qui, par une boucle de rétroaction positive, entraîne la libération de davantage de sérotonine par les plaquettes. Cette boucle amplifie la réponse et contribue ainsi à l’hypertension présente chez les femmes ayant une PE. Précédemment, il a été démontré par notre laboratoire que le Bisphénol A (BPA) s’accumulait davantage dans le placenta des femmes avec PE en comparaison aux femmes normotensives. Cette accumulation pourrait découler d’une perturbation de sa métabolisation qui impliquerait notamment la β-glucuronidase (GUSB). Des études chez les animaux ont quant à elles démontré que le BPA pouvait inhiber l’activité de la monoamine oxydase (MAO) à forte dose. Nous avons étudié l’effet du BPA à faible concentration (10 ng/ml) sur la MAO-A des cellules placentaires et démontré que le BPA inhibait la MAO-A de façon significative sans affecter son expression protéique. Afin d’expliquer l’accumulation particulière du BPA chez les femmes PE, nous avons comparé l’activité spécifique et l’expression protéique de la β-glucuronidase (GUSB) placentaire en utilisant un devis cas-témoins. Une tendance non significative suggère que la GUSB pourrait partiellement contribuer à l’accumulation du BPA chez les femmes PE. Nous avons étudié la relation entre la concentration sérique maternelle de BPA et la concentration à laquelle le fœtus est exposé par régression linéaire et corrélation de Spearman. Un tel modèle ne pourrait être utilisé pour déterminer de façon quantitative l’exposition fœtale. En revanche, en vue de la forte corrélation entre ces deux variables, une haute concentration sérique maternelle de BPA devrait se refléter par une haute exposition fœtale. Cette corrélation implique aussi que le métabolisme placentaire ne joue pas un rôle significatif dans la protection du fœtus. Le BPA pourrait ainsi contribuer à l’hypertension chez les femmes PE présentant une dysfonction endothéliale en inhibant la MAO-A et ainsi, favorisant la hausse de sérotonine circulante. Cette étude suggère les bases d’un mécanisme par lequel le BPA s’accumulerait davantage chez les femmes PE et affecterait ainsi la MAO-A placentaire et potentiellement, la MAO-A fœtale vu ses propriétés physico-chimiques.
Abstract : Preeclampsia (PE) is an hypertensive disorder of pregnancy characterized by a generalized endothelial dysfunction where the response to vasodilatation signals is compromised. The binding of serotonin to its S[subscript 2] receptor subtype 2 releases vasoconstrictor molecules which, by a positive retroaction loop, stimulates the release of more serotonin from platelets. This positive retroaction loop stimulates the vasoconstriction of blood vessels and contributes to the hypertension in women with PE. Previously, we showed that Bisphenol A (BPA) accumulates more in the placenta of women with PE than in normotensive women. This accumulation may be the result of an impaired metabolization due to the action of the β-Glucuronidase (GUSB). Animal studies showed that BPA at high dose could lower the activity of the monoamine oxidase A (MAO-A), an enzyme implicated in the metabolism of serotonin. We studied the impact of BPA at low dose (10 ng/ml) in trophoblastic primary cells and showed that even at low dose, BPA can lower its activity without affecting the protein expression. To determine if GUSB could be the cause of the BPA accumulation in women with PE, we studied its activity and protein expression in placental biopsies from women with and without PE. A nonsignificant tendency showed that the GUSB activity and protein expression were higher in women with PE. To study the impact of placental metabolism in the fetal exposure, we studied the relation between maternal and fetal concentrations of BPA with linear regression analysis and Spearman’s correlation. We showed that maternal BPA could not precisely predict the fetal exposure and that the placental metabolism is probably limited in light of the strong correlation between both variables. This strong correlation also implied that high maternal exposure would result in high fetal exposure. This study shows that the accumulation of BPA in preeclamptic women could contribute to maternal hypertension by interacting with serotonin levels. This accumulation could partially be attributed to a higher GUSB, but other factors are probably implicated. The strong correlation between maternal and fetal exposure implies that the placental metabolism of BPA is limited and does not protect the fetus significantly. This study suggests the basis of a mechanism explaining the abnormal accumulation of BPA in the placenta in women with PE and its impact on the placental MAO-A and potentially, the foetal MAO-A because of its physico-chemical properties.
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Pagès, i. Pi Gemma. "Intrathecal administration of AAVrh10 coding for β‐glucuronidase corrects biochemical and histological hallmarks of mucopolysaccharidosis type VII mice and improves behavior and survival." Doctoral thesis, Universitat Autònoma de Barcelona, 2015. http://hdl.handle.net/10803/300733.

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La mucopolisacaridosi tipus VII (MPS VII) és una malaltia monogènica molt rara inclosa en el grup de malalties lisosòmiques. Està causada per la manca d’activitat β-­‐ glucuronidasa, un enzim lisosòmic involucrat en la via de degradació de glicosaminoglicans. Aquesta alteració congènita causa una disfunció del sistema lisosòmic que provoca una acumulació anormal als lisosomes i la disrupció de l’homeòstasi de la cèl·lula. La malaltia presenta diferents graus de severitat clínica que van des de la mort prenatal fins a pacients amb formes lleus i una esperança de vida de 20 o 30 anys. Entre els casos que presenten un inici postnatal de la malaltia, la forma més severa de MPS VII es caracteritza per hepatomegàlia, esplenomegàlia, anomalies esquelètiques, desenvolupament tardà i retard mental, entre altres símptomes. Actualment els tractaments per als pacients de MPS VII són intervencions per pal·liar els símptomes de la malaltia, però no disposen de cap tractament curatiu. La teràpia gènica és una estratègia prometedora de cara a trobar una cura per a malalties monogèniques. Entre els vectors de teràpia gènica disponibles, els virus adeno-­‐associats (AAV) presenten certes característiques que els fan molt atractius per a la teràpia gènica: permeten la transducció tant de cèl·lules en divisió com quiescents, proporcionen una expressió del transgèn a llarg termini, no poden replicar-­‐se de manera autònoma sense un virus accessori i, a més, les infeccions naturals dels AAV no són patogèniques. Diferents serotips d’AAV s’han utilitzat com a vectors de teràpia gènica en estudis preclínics. Entre ells, els serotips AAV9 i AAVrh10 són els que presenten una capacitat de transducció més gran, així com una gamma més àmplia de tipus cel·lulars que poden transduir, especialment al sistema nerviós central. El serotip AAVrh10 no és d’origen humà. Per això, el fet d’utilitzar-­‐lo com a vector de teràpia gènica podria evitar la neutralització per part dels factors immunològics específics contra AAV, factors que són presents en el sèrum humà després d’infeccions d’AAV naturals. Tanmateix, el reconeixement creuat per part d’anticossos generats contra AAV2, el serotip humà més comú, podrien interferir en el resultat de la teràpia. En aquest treball es proposa una estratègia de teràpia gènica per a MPS VII basada en una única injecció intratecal d’un vector AAVrh10 que conté el gen de la β-­‐ glucuronidasa, testada en ratolins MPS VII adults joves. Es mostra que l’administració del vector al líquid cefaloraquidi (LCR) per punció lumbar, una tècnica molt poc invasiva, permet la transducció d’estructures del sistema nerviós central (SNC) utilitzant una dosi de vector més baixa que mitjançant injecció intravenosa. A més, el drenatge del vector del LCR cap al torrent sanguini comporta la transducció d’òrgans somàtics com el fetge. Això genera una font de β-­‐glucuronidasa que aconsegueix una activitat enzimàtica en sèrum comparable a la de ratolins sans. L’expressió sostinguda de l’enzim recombinant per part de les cèl·lules transduïdes, així com la correcció creuada deguda a la secreció de l’enzim, aconsegueixen la correcció de les característiques bioquímiques i histopatològiques de la malaltia, tant al SNC com als òrgans somàtics. Aquesta correcció a nivell cel·lular condueix a una millora significativa de les capacitats físiques, cognitives i emocionals dels ratolins MPS VII i aconsegueix doblar la seva esperança de vida.
Mucopolysaccharidosis type VII (MPS VII) is an ultrarare monogenic lysosomal storage disease. It is caused by the lack of β-­‐glucuronidase activity, a lysosomal enzyme involved in the degradation pathway of glycosaminoglycans. This inborn genetic alteration causes a dysfunction of the lysosomal system that entails abnormal lysosomal storage and disruption of cell homeostasis. The disease presents a range of clinical severity among patients, from death in utero to a life expectancy of up to 20 or 30 years for the milder forms. The severe form of MPS VII among cases with postnatal disease onset is characterized by hepatosplenomegaly, skeletal abnormalities, developmental delay and mental retardation, among other symptoms. Currently, the only treatments for MPS VII patients are interventions to alleviate the symptoms, but no curative treatment is available. Gene therapy is a promising therapeutic approach to find a cure for monogenic diseases. Among the available gene delivery vectors, adeno-­‐associated viruses (AAVs) present several features that make them attractive for gene therapy strategies: they are able to transduce dividing and quiescent cells, they provide long term expression of the transgene, they are not able to autonomously replicate without a helper virus, and wild type AAV infections are not pathogenic. Different AAV serotypes have been used as gene therapy vectors in preclinical studies. Among them, AAV9 and AAVrh10 are the serotypes which show greater transduction capacity and a broader range of cell-­‐type specific tropism, particularly in the central nervous system. The use of AAVrh10, a non-­‐human serotype, may avoid the neutralization by anti-­‐AAV immune factors present in human sera after natural AAV infections. However, cross-­‐reactivity with antibodies raised against AAV2, the most common human AAV serotype, may still interfere in the therapeutic outcome. In this work, we propose a gene therapy strategy for MPS VII based on a single intrathecal injection of an AAVrh10 coding for the β-­‐glucuronidase gene, tested in young adult MPS VII mice. We show that vector delivery to the CSF by lumbar puncture, a poorly invasive technique, allows the transduction of CNS structures using a lower vector dose than by intravenous delivery. In addition, the drainage of the vector from the CSF to the bloodstream results in transduction of somatic organs such as liver, thus providing a systemic β-­‐glucuronidase source that achieves serum enzymatic activity comparable to wild type mice. The sustained recombinant enzyme expression by AAV-­‐transduced cells, and the cross-­‐correction provided by enzyme secretion, attains the correction of biochemical and histopathological hallmarks of the disease in CNS and somatic organs. This correction at the cellular level leads to a significant improvement of physical, cognitive and emotional characteristics of MPS VII mice and a doubling of the MPS VII mouse life span.
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Sibanda, Ntsako. "Evaluation of high recombinant protein secretion phenotype of saccharomyces cerevisiae segregant." Thesis, University of Limpopo, 2016. http://hdl.handle.net/10386/1803.

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Thesis (MSc. (Biochemistry)) --University of Limpopo, 2016
The ever increasing cost of fossil-based fuels and the accompanying concerns about their impact on the environment is driving research towards clean and renewable sources of energy. Bioethanol has the potential to be a replacement for liquid transportation fuels. In addition to its near zero nett carbon dioxide emissions, bio-ethanol has a high energy to weight ratio and can easily be stored in high volumes. To produce bioethanol at economically competitive prices, the major cost in the production process needs to be addressed. The addition of enzymes to hydrolyse the lignocellulosic fraction of the agricultural waste to simple sugars is considered to be the major contributor to high production cost. A consolidated bioprocess (CBP) which ideally combines all the steps that are currently accomplished in different reactors by different microorganisms into a single process step would be a more economically feasible solution. In this study the potential of yeast hybridization with a CBP approach was used. In order to evaluate the reduction or elimination of the addition of cellulolytic and hemi-cellulolytic enzymes to the ethanol production process. High cellobiohydrolase I secreting progeny from hybridization of an industrial bioethanol yeast strain, S. cerevisiae M0341, and a laboratory strain S. cerevisiae Y294 were isolated. In order to determine if this characteristic was specific to cellobiohydrolase I secretion, these strains were evaluated for their ability to secrete other relevant recombinant hydrolase enzymes for CBP-based ethanol production. A total of seven S. cerevisiae strains were chosen from a progeny pool of 28 supersecreting hybrids and reconstructed to create two parental strains; S. cerevisiae M0341 and S. cerevisiae Y294, together with their hybrid segregants strains H3M1, H3M28, H3H29, H3K27 and H3O23. Three episomal plasmids namely pNS201, pNS202 and pNS203 were constructed; these plasmids together with two already available plasmids, namely pRDH166 and pRDH182 contained genes for different reporter enzymes, namely β-glucosidase I, xylanase II, endoglucanase lll, cellobiohydrolase l and α-glucuronidase. To allow for selection of the episomal plasmids, homologous recombination was used to replace the functional URA3 gene of selected strains, with the non-functional ura3 allele from the Y294 strain. Enzyme activity was used as an indicator of the amount of enzyme secreted. Fermentation studies in a bioreactor were used to determine the metabolic burden imposed on the segregants expressing the cellobiohydrolase at high levels. In addition all segregants were tested for resistance to inhibitors commonly found in pre-treated lignocellulosic material. The M28_Cel7A was found to be the best secretor of Cel7A (Cellobiohydrolase l); however it seems as though this phenomenon imposes a significant metabolic burden on the yeast. The supersecreting hybrid strains cannot tolerate lignocellulosic inhibitors at concentrations commonly produced during pretreatment
The National Research Foundation - Renewable Energy Scholarship (NRF-RSES)
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Book chapters on the topic "Β-Glucuronidases"

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Fishman, William H. "Determination of β-Glucuronidases." In Methods of Biochemical Analysis, 77–145. Hoboken, NJ, USA: John Wiley & Sons, Inc., 2006. http://dx.doi.org/10.1002/9780470110331.ch3.

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Ibrahim, Nasser El-Din, Weilan Shao, and Kesen Ma. "Properties and Biotechnological Applications of β-Glucuronidases." In Biotechnology of Microorganisms, 147–78. Series statement: Innovations in biotechnology; volume 2 |: Apple Academic Press, 2019. http://dx.doi.org/10.1201/9780429434112-7.

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Schomburg, Dietmar, and Ida Schomburg. "baicalin-β-d-glucuronidase 3.2.1.167." In Class 2–3.2 Transferases, Hydrolases, 624–30. Berlin, Heidelberg: Springer Berlin Heidelberg, 2013. http://dx.doi.org/10.1007/978-3-642-36240-8_121.

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Panchal, Dixita, Vrutika Lad, Meonis Pithawala, and Natarajan Amaresan. "Determination of β-Glucuronidase Production." In Methods and Protocols in Food Science, 29–31. New York, NY: Springer US, 2022. http://dx.doi.org/10.1007/978-1-0716-2509-5_3.

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Fishman, W. H. "Assay, Activity and Purification of β-Glucuronidase." In Ciba Foundation Symposium - Steroid Hormones and Enzymes (Book II of Colloquia on Endocrinology, Vol. 1), 229–34. Chichester, UK: John Wiley & Sons, Ltd., 2008. http://dx.doi.org/10.1002/9780470718742.ch1.

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Chapman, V. M., D. R. Miller, E. Novak, and R. W. Elliott. "Alleles of β-Glucuronidase Found in Wild Mice." In The Wild Mouse in Immunology, 114–23. Berlin, Heidelberg: Springer Berlin Heidelberg, 1986. http://dx.doi.org/10.1007/978-3-642-71304-0_14.

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Mills, G. T. "The Nature, Properties and Function of β-Glucuronidase." In Ciba Foundation Symposium - Steroid Hormones and Enzymes (Book II of Colloquia on Endocrinology, Vol. 1), 235–42. Chichester, UK: John Wiley & Sons, Ltd., 2008. http://dx.doi.org/10.1002/9780470718742.ch2.

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Fishman, W. H. "The Effects of Hormones on β-Glucuronidase Activity." In Ciba Foundation Symposium - Steroid Hormones and Enzymes (Book II of Colloquia on Endocrinology, Vol. 1), 257–62. Chichester, UK: John Wiley & Sons, Ltd., 2008. http://dx.doi.org/10.1002/9780470718742.ch5.

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Chapman, Verne M., David Adler, Cesar Labarca, and Linda Wudl. "Genetic Variation of β-Glucuronidase Expression During Early Embryogenesis." In Ciba Foundation Symposium 40 - Embryogenesis in Mammals, 115–31. Chichester, UK: John Wiley & Sons, Ltd., 2008. http://dx.doi.org/10.1002/9780470720226.ch7.

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Béziat, Chloé, Jürgen Kleine-Vehn, and Elena Feraru. "Histochemical Staining of β-Glucuronidase and Its Spatial Quantification." In Methods in Molecular Biology, 73–80. New York, NY: Springer New York, 2016. http://dx.doi.org/10.1007/978-1-4939-6469-7_8.

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Conference papers on the topic "Β-Glucuronidases"

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Gunda, Naga Siva Kumar, Selvaraj Naicker, Maryam S. Ghoraishi, Subir Bhattacharjee, Thomas G. Thundat, and Sushanta K. Mitra. "Microspot With Integrated Wells (MSIW) for the Detection of E.coli." In ASME 2013 11th International Conference on Nanochannels, Microchannels, and Minichannels. American Society of Mechanical Engineers, 2013. http://dx.doi.org/10.1115/icnmm2013-73037.

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There is an increasing problem in getting quality water for developing countries. Water system is contaminated and without proper treatment, it has been consumed as drinking water. It is a big problem for health. Escherichia coli (E.coli) is the main cause for the contamination of water and illness in people. Early detection of E.coli presence in the drinking water followed by subsequent treatment for elimination of E.coli can solve this problem. The present work developed a new method for detecting E.coli in contaminated water using microspot with integrated wells (MSIW). The method involves the fabrication of MSIW, coating the MSIW with enzyme substrates such as 4-MUG substrate (4-Methylumbelliferyl-β-D-glucuronide, trihydrate) and Red-Gal substrate (6-Chloro-3-indolyl-β-D-galactoside) in proper medium and dispensing the contaminated water into MSIW. GlucuronidaseA (gusA) gene in E.coli encodes the beta-D-Glucuronidase (GUS) to hydrolyze the substrate 4-MUG enzymatically which leads to the generation of the fluorigenic compound 4-MU. β-galactosidase enzyme in E.coli produces red color when it reacts with Red-Gal substrate. Using portable optical readers, average color/fluorescence intensity emitting by MSIW is measured and quantified. Comparing obtained intensity values with calibrated intensity values, the level of contamination can be predicted for early warnings.
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Huang, HK, TL Cheng, CH Lin, HC Wu, IS Chen, KH Gan, and HS Chang. "Anti-Escherichia coli β-glucuronidase activity constituents from the root of Neolitsea konishii." In GA 2017 – Book of Abstracts. Georg Thieme Verlag KG, 2017. http://dx.doi.org/10.1055/s-0037-1608150.

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Koschelnik, J., W. Vogl, M. Epp, and M. Lackner. "Rapid analysis of β-D-glucuronidase activity in water using fully automated technology." In WATER RESOURCES MANAGEMENT 2015. Southampton, UK: WIT Press, 2015. http://dx.doi.org/10.2495/wrm150401.

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Caruso, G., G. Zappalà, R. Caruso, and E. Crisafi. "Assessment of Escherichia coli viability in coastal Sicilian waters by fluorescent antibody and β-glucuronidase activity methods." In COASTAL ENVIRONMENT 2006. Southampton, UK: WIT Press, 2006. http://dx.doi.org/10.2495/cenv060061.

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Kunihiro, Andrew, Julia A. Brickey, jennifer B. Frye, Paula B. Luis, Claus Schneider, and Janet L. Funk. "Abstract 3002: Bone-protective curcumin circulates as a pro-drug conjugate that is activated in bone by β-glucuronidase." In Proceedings: AACR Annual Meeting 2019; March 29-April 3, 2019; Atlanta, GA. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/1538-7445.am2019-3002.

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Kunihiro, Andrew, Julia A. Brickey, jennifer B. Frye, Paula B. Luis, Claus Schneider, and Janet L. Funk. "Abstract 3002: Bone-protective curcumin circulates as a pro-drug conjugate that is activated in bone by β-glucuronidase." In Proceedings: AACR Annual Meeting 2019; March 29-April 3, 2019; Atlanta, GA. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/1538-7445.sabcs18-3002.

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Piwowarski, JP, I. Stanisławska, S. Granica, AK Kiss, and T. Moeslinger. "The role of β-glucuronidase in disposition and anti-inflammatory effects of urolithins- gut microbiota-derived metabolites of ellagitannins." In GA 2017 – Book of Abstracts. Georg Thieme Verlag KG, 2017. http://dx.doi.org/10.1055/s-0037-1608033.

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Shin, Dong Yeon, Taeik Jang, Ho Young Song, Sung Min Kim, Yun-Hee Park, Chang Sik Park, Hwanhee Oh, Juyuel Baek, Jeiwook Chae, and Chang-Sun Lee. "Abstract A108: Specific and traceless payload release from HER2-ADC containing LBG-linker prodrug by the action of lysosomal β-glucuronidase." In Abstracts: AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; October 26-30, 2019; Boston, MA. American Association for Cancer Research, 2019. http://dx.doi.org/10.1158/1535-7163.targ-19-a108.

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Simone, E. R., T. A. Davies, N. A. Zabe, S. M. Greenberg-seperaky, and N. E. Larsen. "EARLY PLATELET-THROMBIN RECEPTORS AND THEIR FUNCTIONS." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643730.

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Human platelets possess less than 1000 high affinity [Kd=10-9]and 50-100,000 receptors of lower [Kd=10-7] affinity for o(α-thrombin. The selective derivatization of thrombin with the bifunctional crosslinking agent, DNCO, has enabled us to identify these receptorsvia covalent binding of either active siteinhibited tosyllyslmethylketothrombin (TLCK-T) or active Ctf-thrombin (T).Kinetic studies of the inhibition of the platelet-thrombin response by covalently and noncovalently bound TLCK-T have helped to elucidate the roles of the high and low affinity thrombin receptors. The activation parameters examined were initial membrane depolarization, cytoplasmic alkalinization,dense granule secretion of serotonin and lysosomal secretion of β-glucuronidase.Isolation and characterization of the thrombin receptors after covalent photocoupling of the TLCK-T or active T- were performed after solubilization by gel filtration. The intact, high affinity receptor moiety, a glycoprotein, has an approximate molecular weight of∽lSO.OOO daltons; occasionally this protein is found as a dimer of ∽360,000 daltons. When exposed to o(α-T the receptor undergoes proteolysis, leaving a protein of∽80,000 daltons and releasing the remaining glycoprotein into the medium.Higher doses of active T have been shown to bind with lower affinity to a larger protein of approximate molecular weight 600,000 daltons anda smaller protein of 46,000 daltons. Both proteins are nonsusceptible to thrombin proteolysis. Reduction and alkylation of the600,000 dalton complex yielded two and possibly three high molecular weight components (200,000, 160,000, and possibly 145,000daltons) which may correspond to previously suggested GP-Ia and GP-Ib of the GP-I complex. Under different solubilization conditions, two other membrane proteins have been found to be part of the GP-I complex; one which is not a glycoprotein, GP-Ic, while the other is associated with the glycocalyx and is called glycocalicin. Glycocalicin and GP-Icdo inhibit thrombin binding,implying that the low affinity receptor is indeed the previously suggested GP-I complex and does not appear to be directly involved withplatelet activation.Examination of the effect of dose and duration of incubation with non-covalently binding TLCK-T on subsequent α-thrombin response suggests the existence of positive cooperativity among thrombin receptors.Although TLCK-T has the same affinity for platelets (Kd) as T , the rateof binding and therefore that of dissociation are lower. Thus for incubation times of 1 minute or less with up to a 2x saturating TLCK-T dose, the subsequent depolarization response to a saturating T dose was enhanced. Exposure to higher TLCK-T (5x saturating)doses led to significant inhibition.Verification of the potentiation observed in noncovalent TLCK-T studies was performed using TLCK-T covalently bound to the platelet receptor with DNCO. Several hundred thrombin molecules were bound to the platelet when a subsaturating dose of TLCK-T(0.0025 U/ml) was used to crosslink, whileseveral thousand resulted with a saturating (0.05 U/ml) TLCK-T dose. Positive cooperativity was observed with low αT doses (0.005 U/ml) when several hundred high affinity receptors are blocked. The parameters studied which exhibited this positive cooperativity were depolarization, pH change and serotonin secretion, α-Glucuronidase secretion was normal. The presence and degreeof enhancement were donor-variableand suggest different threshhold thrombin dose requirements. The enhancement observed can be attributed to either an increased rate of binding (increased affinity) or to an increased number of exposed binding sites. Since little difference was found between the number of TLCK-T molecules bound after30 versus 60 seconds, we conclude that thepotentiation is more likely due to an increased number of exposed binding sites. Results from covalent crosslinks using a fluorescein and rhodamine labeled-TLCK-T and the fluorescence activated cellsorter support this hypothesis. The sensitization of the high affinity binding sitesby partial occupancy implies these bindingsites are responsible for depolarization, pH change and dense granule secretion (the rapid initial activation response), while βglucuronidase secretion, a secondary response, is otherwise controlled.
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