Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells
Reporter genes are important tools in many biological disciplines. The discovery of novel reporter genes is relatively rare. However, known reporter genes are constantly applied to novel applications. This study reports the performance of the bilirubin-dependent fluorescent protein UnaG from the Jap...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Biosensors |
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| Online Access: | https://www.mdpi.com/2079-6374/13/2/232 |
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| author | Céline S. M. Richard Hymonti Dey Frode Øyen Munazza Maqsood Hans-Matti Blencke |
| author_facet | Céline S. M. Richard Hymonti Dey Frode Øyen Munazza Maqsood Hans-Matti Blencke |
| author_sort | Céline S. M. Richard |
| collection | DOAJ |
| description | Reporter genes are important tools in many biological disciplines. The discovery of novel reporter genes is relatively rare. However, known reporter genes are constantly applied to novel applications. This study reports the performance of the bilirubin-dependent fluorescent protein UnaG from the Japanese eel <i>Anguilla japonicas</i> in live <i>Escherichia coli</i> cells in response to the disruption of outer membrane (OM) integrity at low bilirubin (BR) concentrations. Using the <i>E. coli</i> wild-type strain MC4100, its isogenic OM-deficient mutant strain NR698, and different OM-active compounds, we show that BR uptake and UnaG fluorescence depend on a leaky OM at concentrations of 10 µM BR and below, while fluorescence is mostly OM integrity-independent at concentrations above 50 µM BR. We suggest that these properties of the UnaG–BR couple might be applied as a biosensor as an alternative to the OM integrity assays currently in use. |
| first_indexed | 2024-03-11T09:04:50Z |
| format | Article |
| id | doaj.art-3eee5b1e4897459ab0ba212cf0eba685 |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-11T09:04:50Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-3eee5b1e4897459ab0ba212cf0eba6852023-11-16T19:25:57ZengMDPI AGBiosensors2079-63742023-02-0113223210.3390/bios13020232Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> CellsCéline S. M. Richard0Hymonti Dey1Frode Øyen2Munazza Maqsood3Hans-Matti Blencke4The Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, NorwayThe Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, NorwayThe Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, NorwayThe Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, NorwayThe Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, NorwayReporter genes are important tools in many biological disciplines. The discovery of novel reporter genes is relatively rare. However, known reporter genes are constantly applied to novel applications. This study reports the performance of the bilirubin-dependent fluorescent protein UnaG from the Japanese eel <i>Anguilla japonicas</i> in live <i>Escherichia coli</i> cells in response to the disruption of outer membrane (OM) integrity at low bilirubin (BR) concentrations. Using the <i>E. coli</i> wild-type strain MC4100, its isogenic OM-deficient mutant strain NR698, and different OM-active compounds, we show that BR uptake and UnaG fluorescence depend on a leaky OM at concentrations of 10 µM BR and below, while fluorescence is mostly OM integrity-independent at concentrations above 50 µM BR. We suggest that these properties of the UnaG–BR couple might be applied as a biosensor as an alternative to the OM integrity assays currently in use.https://www.mdpi.com/2079-6374/13/2/232reporter genesynthetic biologyUnaGouter membranebilirubinbiosensor |
| spellingShingle | Céline S. M. Richard Hymonti Dey Frode Øyen Munazza Maqsood Hans-Matti Blencke Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells Biosensors reporter gene synthetic biology UnaG outer membrane bilirubin biosensor |
| title | Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells |
| title_full | Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells |
| title_fullStr | Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells |
| title_full_unstemmed | Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells |
| title_short | Outer Membrane Integrity-Dependent Fluorescence of the Japanese Eel UnaG Protein in Live <i>Escherichia coli</i> Cells |
| title_sort | outer membrane integrity dependent fluorescence of the japanese eel unag protein in live i escherichia coli i cells |
| topic | reporter gene synthetic biology UnaG outer membrane bilirubin biosensor |
| url | https://www.mdpi.com/2079-6374/13/2/232 |
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