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Teriosina, A, Barsukov, IL, Cartmell, A, Powell, AK, Stachulski, AV and Yates, EA (2025) Detection of β-d-glucuronidase activity in environmental samples using 4-fluorophenyl β-d-glucuronide and 19F NMR. Analytical Methods. ISSN 1759-9660
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Abstract
Common methods for establishing the presence of enteric bacteria polluting water supplies, or in other samples, rely on detecting the hydrolysis of model glucuronide substrates by glucuronidases to release a phenolic product quantifiable by absorbance or fluorescence. Substrates include the β-D-glucuronides of p-nitrophenol, and umbelliferyl or quercetin derivatives. One limitation is that it may be difficult or impossible to quantify the released phenolic moiety in samples that are strongly coloured or, that contain fluorescent compounds. Exploiting the sensitivity available from the 19F nucleus to changes in chemical environment which can be detected by 19F NMR spectroscopy, and the almost complete absence of 19F from naturally-occurring samples containing organic matter, which provides background-free signals, we propose a model substrate; 4-fluorophenyl β-D-glucuronide (4FP-glucuronide). The 19F NMR chemical shift position of 4FP-glucuronide changes from −121.0 ppm upon hydrolysis to release 4-fluorophenol, at −124.9 ppm (at pH 6.8), enabling detection of β-glucuronidase activity. We illustrate the use of this substrate with environmental samples from forest soil, standing water, and mud from cattle pasture. Each of these would challenge conventional methods, owing to their opacity or the presence of coloured organic material. The technique enables detection of glucuronidases, a widely-used proxy for enteric bacteria, extending the scope of testing beyond water to include environmental and other challenging samples.
| Item Type: | Article |
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| Uncontrolled Keywords: | 40 Engineering; 4018 Nanotechnology; 0301 Analytical Chemistry; 0399 Other Chemical Sciences; 3401 Analytical chemistry; 4004 Chemical engineering |
| Subjects: | Q Science > QD Chemistry Q Science > QH Natural history > QH301 Biology |
| Divisions: | Pharmacy and Biomolecular Sciences |
| Publisher: | Royal Society of Chemistry (RSC) |
| SWORD Depositor: | A Symplectic |
| Date Deposited: | 14 Feb 2025 16:57 |
| Last Modified: | 14 Feb 2025 17:00 |
| DOI or ID number: | 10.1039/d4ay01723d |
| URI: | https://researchonline.ljmu.ac.uk/id/eprint/25648 |
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