Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02
Cell-based biosensors harness a cell’s ability to respond to the environment by repurposing its sensing mechanisms. MerR family proteins are activator/repressor switches that regulate the expression of bacterial metal resistance genes and have been used in metal biosensors. Upon metal binding, a con...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-01-01
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| Series: | Micromachines |
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| Online Access: | https://www.mdpi.com/2072-666X/12/2/142 |
| _version_ | 1797406016532381696 |
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| author | Georgina Baya Stephen Muhindi Valentine Ngendahimana Jonathan Caguiat |
| author_facet | Georgina Baya Stephen Muhindi Valentine Ngendahimana Jonathan Caguiat |
| author_sort | Georgina Baya |
| collection | DOAJ |
| description | Cell-based biosensors harness a cell’s ability to respond to the environment by repurposing its sensing mechanisms. MerR family proteins are activator/repressor switches that regulate the expression of bacterial metal resistance genes and have been used in metal biosensors. Upon metal binding, a conformational change switches gene expression from off to on. The genomes of the multimetal resistant bacterial strains, <i>Stenotrophomonas maltophilia</i> Oak Ridge strain 02 (<i>S. maltophilia</i> 02) and <i>Enterobacter</i> sp. YSU, were recently sequenced. Sequence analysis and gene cloning identified three mercury resistance operons and three MerR switches in these strains. Transposon mutagenesis and sequence analysis identified <i>Enterobacter</i> sp. YSU zinc and copper resistance operons, which appear to be regulated by the protein switches, ZntR and CueR, respectively. Sequence analysis and reverse transcriptase polymerase chain reaction (RT-PCR) showed that a CueR switch appears to activate a <i>S. maltophilia</i> 02 copper transport gene in the presence of CuSO<sub>4</sub> and HAuCl<sub>4</sub>·3H<sub>2</sub>O. In previous studies, genetic engineering replaced metal resistance genes with the reporter genes for β-galactosidase, luciferase or the green fluorescence protein (GFP). These produce a color change of a reagent, produce light, or fluoresce in the presence of ultraviolet (UV) light, respectively. Coupling these discovered operons with reporter genes has the potential to create whole-cell biosensors for HgCl<sub>2</sub>, ZnCl<sub>2</sub>, CuSO<sub>4</sub> and HAuCl<sub>4</sub>·3H<sub>2</sub>O. |
| first_indexed | 2024-03-09T03:20:05Z |
| format | Article |
| id | doaj.art-2058ef6c836e48f4aa3d69bc3ec13d2c |
| institution | Directory Open Access Journal |
| issn | 2072-666X |
| language | English |
| last_indexed | 2024-03-09T03:20:05Z |
| publishDate | 2021-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Micromachines |
| spelling | doaj.art-2058ef6c836e48f4aa3d69bc3ec13d2c2023-12-03T15:11:25ZengMDPI AGMicromachines2072-666X2021-01-0112214210.3390/mi12020142Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02Georgina Baya0Stephen Muhindi1Valentine Ngendahimana2Jonathan Caguiat3Department of Biological and Chemical Sciences, Youngstown State University, Youngstown, OH 44555, USADepartment of Biological Sciences, University of Toledo, Toledo, OH 43606, USABiology Department, Lone Star College-CyFair, 9191 Barker Cypress Rd, Cypress, TX 77433, USADepartment of Biological and Chemical Sciences, Youngstown State University, Youngstown, OH 44555, USACell-based biosensors harness a cell’s ability to respond to the environment by repurposing its sensing mechanisms. MerR family proteins are activator/repressor switches that regulate the expression of bacterial metal resistance genes and have been used in metal biosensors. Upon metal binding, a conformational change switches gene expression from off to on. The genomes of the multimetal resistant bacterial strains, <i>Stenotrophomonas maltophilia</i> Oak Ridge strain 02 (<i>S. maltophilia</i> 02) and <i>Enterobacter</i> sp. YSU, were recently sequenced. Sequence analysis and gene cloning identified three mercury resistance operons and three MerR switches in these strains. Transposon mutagenesis and sequence analysis identified <i>Enterobacter</i> sp. YSU zinc and copper resistance operons, which appear to be regulated by the protein switches, ZntR and CueR, respectively. Sequence analysis and reverse transcriptase polymerase chain reaction (RT-PCR) showed that a CueR switch appears to activate a <i>S. maltophilia</i> 02 copper transport gene in the presence of CuSO<sub>4</sub> and HAuCl<sub>4</sub>·3H<sub>2</sub>O. In previous studies, genetic engineering replaced metal resistance genes with the reporter genes for β-galactosidase, luciferase or the green fluorescence protein (GFP). These produce a color change of a reagent, produce light, or fluoresce in the presence of ultraviolet (UV) light, respectively. Coupling these discovered operons with reporter genes has the potential to create whole-cell biosensors for HgCl<sub>2</sub>, ZnCl<sub>2</sub>, CuSO<sub>4</sub> and HAuCl<sub>4</sub>·3H<sub>2</sub>O.https://www.mdpi.com/2072-666X/12/2/142whole-cell biosensorMerR family proteinZntRCueRbacterial metal resistanceHgCl<sub>2</sub> |
| spellingShingle | Georgina Baya Stephen Muhindi Valentine Ngendahimana Jonathan Caguiat Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 Micromachines whole-cell biosensor MerR family protein ZntR CueR bacterial metal resistance HgCl<sub>2</sub> |
| title | Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 |
| title_full | Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 |
| title_fullStr | Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 |
| title_full_unstemmed | Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 |
| title_short | Potential Whole-Cell Biosensors for Detection of Metal Using MerR Family Proteins from <i>Enterobacter</i> sp. YSU and <i>Stenotrophomonas maltophilia</i> OR02 |
| title_sort | potential whole cell biosensors for detection of metal using merr family proteins from i enterobacter i sp ysu and i stenotrophomonas maltophilia i or02 |
| topic | whole-cell biosensor MerR family protein ZntR CueR bacterial metal resistance HgCl<sub>2</sub> |
| url | https://www.mdpi.com/2072-666X/12/2/142 |
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