Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>)
Bacterial infections in fish farms increase mass mortality and rapid detection of infection can help prevent its widespread. Lactate is an important biomarker for early diagnosis of bacterial infections in farmed olive flounder (<i>Paralichthys olivaceus</i>). To determine the lactate le...
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MDPI AG
2021-11-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/11/11/439 |
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| author | Thenmozhi Rajarathinam Seonghye Kim Dinakaran Thirumalai Sujin Lee Minho Kwon Hyun-jong Paik Suhkmann Kim Seung-Cheol Chang |
| author_facet | Thenmozhi Rajarathinam Seonghye Kim Dinakaran Thirumalai Sujin Lee Minho Kwon Hyun-jong Paik Suhkmann Kim Seung-Cheol Chang |
| author_sort | Thenmozhi Rajarathinam |
| collection | DOAJ |
| description | Bacterial infections in fish farms increase mass mortality and rapid detection of infection can help prevent its widespread. Lactate is an important biomarker for early diagnosis of bacterial infections in farmed olive flounder (<i>Paralichthys olivaceus</i>). To determine the lactate levels, we designed a disposable amperometric biosensor based on Prussian blue nanozyme and lactate oxidase (LOX) entrapped in copolymer-reduced graphene oxide (P-rGO) on screen-printed carbon electrodes. Because LOX is inherently unstable, P-rGO nanosheets were utilized as a base matrix to immobilize it. After optimization in terms of enzyme loading, operating potential, and pH, the biosensor displayed maximum current responses within 5 s at the applied potential of –0.1 V vs. internal Ag/AgCl. The biosensor had Langmuir-type response in the lactate concentration range from 10 µM to 1.6 mM, a dynamic linear response range of 10–100 µM, a sensitivity of 15.9 µA mM<sup>−1</sup> cm<sup>−2</sup>, and a lower detection limit of 3.1 µM (S/N = 3). Additionally, the biosensor featured high reproducibility, good selectivity, and stability till four weeks. Its practical applicability was tested in olive flounder infected by <i>Streptococcus parauberis</i> against the uninfected control. The results were satisfactory compared to those of a standard colorimetric assay kit, validating our method. |
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| last_indexed | 2024-03-10T05:39:55Z |
| publishDate | 2021-11-01 |
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| series | Biosensors |
| spelling | doaj.art-092b93a50d6c48f7b818b4f7cd0807d72023-11-22T22:36:11ZengMDPI AGBiosensors2079-63742021-11-01111143910.3390/bios11110439Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>)Thenmozhi Rajarathinam0Seonghye Kim1Dinakaran Thirumalai2Sujin Lee3Minho Kwon4Hyun-jong Paik5Suhkmann Kim6Seung-Cheol Chang7Department of Cogno-Mechatronics Engineering, College of Nanoscience and Nanotechnology, Pusan National University, Busan 46241, KoreaDepartment of Chemistry, Pusan National University, Busan 46241, KoreaDepartment of Cogno-Mechatronics Engineering, College of Nanoscience and Nanotechnology, Pusan National University, Busan 46241, KoreaDepartment of Chemistry, Pusan National University, Busan 46241, KoreaDepartment of Polymer Science and Engineering, Pusan National University, Busan 46241, KoreaDepartment of Polymer Science and Engineering, Pusan National University, Busan 46241, KoreaDepartment of Chemistry, Pusan National University, Busan 46241, KoreaDepartment of Cogno-Mechatronics Engineering, College of Nanoscience and Nanotechnology, Pusan National University, Busan 46241, KoreaBacterial infections in fish farms increase mass mortality and rapid detection of infection can help prevent its widespread. Lactate is an important biomarker for early diagnosis of bacterial infections in farmed olive flounder (<i>Paralichthys olivaceus</i>). To determine the lactate levels, we designed a disposable amperometric biosensor based on Prussian blue nanozyme and lactate oxidase (LOX) entrapped in copolymer-reduced graphene oxide (P-rGO) on screen-printed carbon electrodes. Because LOX is inherently unstable, P-rGO nanosheets were utilized as a base matrix to immobilize it. After optimization in terms of enzyme loading, operating potential, and pH, the biosensor displayed maximum current responses within 5 s at the applied potential of –0.1 V vs. internal Ag/AgCl. The biosensor had Langmuir-type response in the lactate concentration range from 10 µM to 1.6 mM, a dynamic linear response range of 10–100 µM, a sensitivity of 15.9 µA mM<sup>−1</sup> cm<sup>−2</sup>, and a lower detection limit of 3.1 µM (S/N = 3). Additionally, the biosensor featured high reproducibility, good selectivity, and stability till four weeks. Its practical applicability was tested in olive flounder infected by <i>Streptococcus parauberis</i> against the uninfected control. The results were satisfactory compared to those of a standard colorimetric assay kit, validating our method.https://www.mdpi.com/2079-6374/11/11/439amperometric biosensormetabolic biomarkerlactatenanozyme–enzyme nanosheets<i>Streptococcus parauberis</i> |
| spellingShingle | Thenmozhi Rajarathinam Seonghye Kim Dinakaran Thirumalai Sujin Lee Minho Kwon Hyun-jong Paik Suhkmann Kim Seung-Cheol Chang Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) Biosensors amperometric biosensor metabolic biomarker lactate nanozyme–enzyme nanosheets <i>Streptococcus parauberis</i> |
| title | Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) |
| title_full | Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) |
| title_fullStr | Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) |
| title_full_unstemmed | Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) |
| title_short | Robust Nanozyme-Enzyme Nanosheets-Based Lactate Biosensor for Diagnosing Bacterial Infection in Olive Flounder (<i>Paralichthys olivaceus</i>) |
| title_sort | robust nanozyme enzyme nanosheets based lactate biosensor for diagnosing bacterial infection in olive flounder i paralichthys olivaceus i |
| topic | amperometric biosensor metabolic biomarker lactate nanozyme–enzyme nanosheets <i>Streptococcus parauberis</i> |
| url | https://www.mdpi.com/2079-6374/11/11/439 |
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