Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species
Glucuronides hydrolysis by intestinal microbial <i>β</i>-Glucuronidases (GUS) is an important procedure for many endogenous and exogenous compounds. The purpose of this study is to determine the impact of experimental conditions on glucuronide hydrolysis by intestinal microbial GUS. Stan...
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MDPI AG
2021-07-01
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| Series: | Pharmaceutics |
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| Online Access: | https://www.mdpi.com/1999-4923/13/7/1043 |
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| author | Christabel Ebuzoeme Imoh Etim Autumn Ikimi Jamie Song Ting Du Ming Hu Dong Liang Song Gao |
| author_facet | Christabel Ebuzoeme Imoh Etim Autumn Ikimi Jamie Song Ting Du Ming Hu Dong Liang Song Gao |
| author_sort | Christabel Ebuzoeme |
| collection | DOAJ |
| description | Glucuronides hydrolysis by intestinal microbial <i>β</i>-Glucuronidases (GUS) is an important procedure for many endogenous and exogenous compounds. The purpose of this study is to determine the impact of experimental conditions on glucuronide hydrolysis by intestinal microbial GUS. Standard probe 4-Nitrophenyl <i>β</i>-D-glucopyranoside (pNPG) and a natural glucuronide wogonoside were used as the model compounds. Feces collection time, buffer conditions, interindividual, and species variations were evaluated by incubating the substrates with enzymes. The relative reaction activity of pNPG, reaction rates, and reaction kinetics for wogonoside were calculated. Fresh feces showed the highest hydrolysis activities. Sonication increased total protein yield during enzyme preparation. The pH of the reaction system increased the activity in 0.69–1.32-fold, 2.9–12.9-fold, and 0.28–1.56-fold for mouse, rat, and human at three different concentrations of wogonoside, respectively. The Vmax for wogonoside hydrolysis was 2.37 ± 0.06, 4.48 ± 0.11, and 5.17 ± 0.16 μmol/min/mg and Km was 6.51 ± 0.71, 3.04 ± 0.34, and 0.34 ± 0.047 μM for mouse, rat, and human, respectively. The inter-individual difference was significant (4–6-fold) using inbred rats as the model animal. Fresh feces should be used to avoid activity loss and sonication should be utilized in enzyme preparation to increase hydrolysis activity. The buffer pH should be appropriate according to the species. Inter-individual and species variations were significant. |
| first_indexed | 2024-03-10T09:28:11Z |
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| issn | 1999-4923 |
| language | English |
| last_indexed | 2024-03-10T09:28:11Z |
| publishDate | 2021-07-01 |
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| series | Pharmaceutics |
| spelling | doaj.art-13e3bf6749f442b683e52a56a786b7262023-11-22T04:41:09ZengMDPI AGPharmaceutics1999-49232021-07-01137104310.3390/pharmaceutics13071043Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and SpeciesChristabel Ebuzoeme0Imoh Etim1Autumn Ikimi2Jamie Song3Ting Du4Ming Hu5Dong Liang6Song Gao7Department of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USADepartment of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USADepartment of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USACollege of Liberal Arts, The University of Texas at Austin, Austin, TX 78712, USADepartment of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USADepartment of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 4901 Calhoun Street, Houston, TX 77204, USADepartment of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USADepartment of Pharmaceutical Science, College of Pharmacy and Health Sciences, Texas Southern University, 3100 Cleburne Street, Houston, TX 77004, USAGlucuronides hydrolysis by intestinal microbial <i>β</i>-Glucuronidases (GUS) is an important procedure for many endogenous and exogenous compounds. The purpose of this study is to determine the impact of experimental conditions on glucuronide hydrolysis by intestinal microbial GUS. Standard probe 4-Nitrophenyl <i>β</i>-D-glucopyranoside (pNPG) and a natural glucuronide wogonoside were used as the model compounds. Feces collection time, buffer conditions, interindividual, and species variations were evaluated by incubating the substrates with enzymes. The relative reaction activity of pNPG, reaction rates, and reaction kinetics for wogonoside were calculated. Fresh feces showed the highest hydrolysis activities. Sonication increased total protein yield during enzyme preparation. The pH of the reaction system increased the activity in 0.69–1.32-fold, 2.9–12.9-fold, and 0.28–1.56-fold for mouse, rat, and human at three different concentrations of wogonoside, respectively. The Vmax for wogonoside hydrolysis was 2.37 ± 0.06, 4.48 ± 0.11, and 5.17 ± 0.16 μmol/min/mg and Km was 6.51 ± 0.71, 3.04 ± 0.34, and 0.34 ± 0.047 μM for mouse, rat, and human, respectively. The inter-individual difference was significant (4–6-fold) using inbred rats as the model animal. Fresh feces should be used to avoid activity loss and sonication should be utilized in enzyme preparation to increase hydrolysis activity. The buffer pH should be appropriate according to the species. Inter-individual and species variations were significant.https://www.mdpi.com/1999-4923/13/7/1043microfloraglucuronideshydrolysismicrobial <i>β</i>-Glucuronidases (GUS) |
| spellingShingle | Christabel Ebuzoeme Imoh Etim Autumn Ikimi Jamie Song Ting Du Ming Hu Dong Liang Song Gao Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species Pharmaceutics microflora glucuronides hydrolysis microbial <i>β</i>-Glucuronidases (GUS) |
| title | Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species |
| title_full | Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species |
| title_fullStr | Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species |
| title_full_unstemmed | Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species |
| title_short | Glucuronides Hydrolysis by Intestinal Microbial <i>β</i>-Glucuronidases (GUS) Is Affected by Sampling, Enzyme Preparation, Buffer pH, and Species |
| title_sort | glucuronides hydrolysis by intestinal microbial i β i glucuronidases gus is affected by sampling enzyme preparation buffer ph and species |
| topic | microflora glucuronides hydrolysis microbial <i>β</i>-Glucuronidases (GUS) |
| url | https://www.mdpi.com/1999-4923/13/7/1043 |
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