A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i>
Arsenic (As) pollution is a widespread problem worldwide. In recent years, biosensors based on enzymatic inhibition have been developed for arsenic detection, making the study of the effect of inhibitors on the selected enzymatic activity crucial for their setup. The arsenate reductase of <i>T...
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MDPI AG
2022-03-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/6/2942 |
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| author | Rosanna Puopolo Giovanni Gallo Danila Limauro Patrizia Contursi Gabriella Fiorentino |
| author_facet | Rosanna Puopolo Giovanni Gallo Danila Limauro Patrizia Contursi Gabriella Fiorentino |
| author_sort | Rosanna Puopolo |
| collection | DOAJ |
| description | Arsenic (As) pollution is a widespread problem worldwide. In recent years, biosensors based on enzymatic inhibition have been developed for arsenic detection, making the study of the effect of inhibitors on the selected enzymatic activity crucial for their setup. The arsenate reductase of <i>Thermus thermophilus</i> HB27, <i>Tt</i>ArsC, reduces As(V) into As(III), but is also endowed with phosphatase activity. This work investigates the inhibitory effects of As(V) and As(III) on phosphatase activity by taking advantage of a simple colorimetric assay; the results show that both of them are non-competitive inhibitors affecting the Vmax but not the K<sub>M</sub> of the reaction. However, their Ki values are different from each other (15.2 ± 1.6 μM for As(V) and 394.4 ± 40.3 µm with As(III)), indicating a higher inhibitory effect by As(V). Moreover, the inhibition-based biosystem results to be selective for As(V) since several other metal ions and salts do not affect <i>Tt</i>ArsC phosphatase activity; it exhibits a sensitivity of 0.53 ± 0.03 mU/mg/μM and a limit of detection (LOD) of 0.28 ± 0.02 μM. The good sensitivity and specificity for As(V) point to consider inhibition of <i>Tt</i>ArsC phosphatase activity for the setup of a novel biosensor for the detection of As(V). |
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| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T19:43:48Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d9fdbdd52ba341c6a4218a58b23806e62023-11-24T01:29:30ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-03-01236294210.3390/ijms23062942A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i>Rosanna Puopolo0Giovanni Gallo1Danila Limauro2Patrizia Contursi3Gabriella Fiorentino4Department of Biology, University of Naples Federico II, Via Cinthia 21, 80126 Napoli, ItalyDepartment of Biology, University of Naples Federico II, Via Cinthia 21, 80126 Napoli, ItalyDepartment of Biology, University of Naples Federico II, Via Cinthia 21, 80126 Napoli, ItalyDepartment of Biology, University of Naples Federico II, Via Cinthia 21, 80126 Napoli, ItalyDepartment of Biology, University of Naples Federico II, Via Cinthia 21, 80126 Napoli, ItalyArsenic (As) pollution is a widespread problem worldwide. In recent years, biosensors based on enzymatic inhibition have been developed for arsenic detection, making the study of the effect of inhibitors on the selected enzymatic activity crucial for their setup. The arsenate reductase of <i>Thermus thermophilus</i> HB27, <i>Tt</i>ArsC, reduces As(V) into As(III), but is also endowed with phosphatase activity. This work investigates the inhibitory effects of As(V) and As(III) on phosphatase activity by taking advantage of a simple colorimetric assay; the results show that both of them are non-competitive inhibitors affecting the Vmax but not the K<sub>M</sub> of the reaction. However, their Ki values are different from each other (15.2 ± 1.6 μM for As(V) and 394.4 ± 40.3 µm with As(III)), indicating a higher inhibitory effect by As(V). Moreover, the inhibition-based biosystem results to be selective for As(V) since several other metal ions and salts do not affect <i>Tt</i>ArsC phosphatase activity; it exhibits a sensitivity of 0.53 ± 0.03 mU/mg/μM and a limit of detection (LOD) of 0.28 ± 0.02 μM. The good sensitivity and specificity for As(V) point to consider inhibition of <i>Tt</i>ArsC phosphatase activity for the setup of a novel biosensor for the detection of As(V).https://www.mdpi.com/1422-0067/23/6/2942arsenicthermostable arsenate reductase<i>Thermus thermophilus</i>biosensor |
| spellingShingle | Rosanna Puopolo Giovanni Gallo Danila Limauro Patrizia Contursi Gabriella Fiorentino A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> International Journal of Molecular Sciences arsenic thermostable arsenate reductase <i>Thermus thermophilus</i> biosensor |
| title | A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> |
| title_full | A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> |
| title_fullStr | A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> |
| title_full_unstemmed | A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> |
| title_short | A New Strategy for As(V) Biosensing Based on the Inhibition of the Phosphatase Activity of the Arsenate Reductase from <i>Thermus thermophilus</i> |
| title_sort | new strategy for as v biosensing based on the inhibition of the phosphatase activity of the arsenate reductase from i thermus thermophilus i |
| topic | arsenic thermostable arsenate reductase <i>Thermus thermophilus</i> biosensor |
| url | https://www.mdpi.com/1422-0067/23/6/2942 |
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