Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper
Over the past decade, copper (Cu) has been recognized as a crucial metal in the differential expression of soluble (sMMO) and particulate (pMMO) forms of methane monooxygenase (MMO) through a mechanism referred to as the “Cu switch”. In this study, we used <i>Methylosinus trichosporium</i&g...
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| Format: | Article |
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MDPI AG
2024-02-01
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| Series: | Methane |
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| Online Access: | https://www.mdpi.com/2674-0389/3/1/7 |
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| author | Dipayan Samanta Tanvi Govil Priya Saxena Lee Krumholz Venkataramana Gadhamshetty Kian Mau Goh Rajesh K. Sani |
| author_facet | Dipayan Samanta Tanvi Govil Priya Saxena Lee Krumholz Venkataramana Gadhamshetty Kian Mau Goh Rajesh K. Sani |
| author_sort | Dipayan Samanta |
| collection | DOAJ |
| description | Over the past decade, copper (Cu) has been recognized as a crucial metal in the differential expression of soluble (sMMO) and particulate (pMMO) forms of methane monooxygenase (MMO) through a mechanism referred to as the “Cu switch”. In this study, we used <i>Methylosinus trichosporium</i> OB3b as a model bacterium to investigate the range of Cu concentrations that trigger the expression of sMMO to pMMO and its effect on growth and methane oxidation. The Cu switch was found to be regulated within Cu concentrations from 3 to 5 µM, with a strict increase in the methane consumption rates from 3.09 to 3.85 µM occurring on the 6th day. Our findings indicate that there was a decrease in the fold changes in the expression of methanobactin (Mbn) synthesis gene (<i>mbnA</i>) with a higher Cu concentration, whereas the Ton-B siderophore receptor gene (<i>mbnT</i>) showed upregulation at all Cu concentrations. Furthermore, the upregulation of the di-heme enzyme at concentrations above 5 µM Cu may play a crucial role in the copper switch by increasing oxygen consumption; however, the role has yet not been elucidated. We developed a quantitative assay based on the naphthalene–Molisch principle to distinguish between the sMMO- and pMMO-expressing cells, which coincided with the regulation profile of the sMMO and pMMO genes. At 0 and 3 µM Cu, the naphthol concentration was higher (8.1 and 4.2 µM, respectively) and gradually decreased to 0 µM naphthol when pMMO was expressed and acted as the sole methane oxidizer at concentrations above 5 µM Cu. Using physical protein–protein interaction, we identified seven transporters, three cell wall biosynthesis or degradation proteins, Cu resistance operon proteins, and 18 hypothetical proteins that may be involved in Cu toxicity and homeostasis. These findings shed light on the key regulatory genes of the Cu switch that will have potential implications for bioremediation and biotechnology applications. |
| first_indexed | 2024-04-24T18:00:08Z |
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| id | doaj.art-21ff1b2588cb4ae19d950ac527d38e5f |
| institution | Directory Open Access Journal |
| issn | 2674-0389 |
| language | English |
| last_indexed | 2024-04-24T18:00:08Z |
| publishDate | 2024-02-01 |
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| series | Methane |
| spelling | doaj.art-21ff1b2588cb4ae19d950ac527d38e5f2024-03-27T13:54:39ZengMDPI AGMethane2674-03892024-02-013110312110.3390/methane3010007Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to CopperDipayan Samanta0Tanvi Govil1Priya Saxena2Lee Krumholz3Venkataramana Gadhamshetty4Kian Mau Goh5Rajesh K. Sani6Department of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USADepartment of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USADepartment of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USADepartment of Microbiology and Plant Biology, University of Oklahoma, Norman, OK 73019, USADepartment of Civil and Environmental Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USAFaculty of Science, Universiti Teknologi Malaysia, Skudai 81310, Johor, MalaysiaDepartment of Chemical and Biological Engineering, South Dakota School of Mines and Technology, Rapid City, SD 57701, USAOver the past decade, copper (Cu) has been recognized as a crucial metal in the differential expression of soluble (sMMO) and particulate (pMMO) forms of methane monooxygenase (MMO) through a mechanism referred to as the “Cu switch”. In this study, we used <i>Methylosinus trichosporium</i> OB3b as a model bacterium to investigate the range of Cu concentrations that trigger the expression of sMMO to pMMO and its effect on growth and methane oxidation. The Cu switch was found to be regulated within Cu concentrations from 3 to 5 µM, with a strict increase in the methane consumption rates from 3.09 to 3.85 µM occurring on the 6th day. Our findings indicate that there was a decrease in the fold changes in the expression of methanobactin (Mbn) synthesis gene (<i>mbnA</i>) with a higher Cu concentration, whereas the Ton-B siderophore receptor gene (<i>mbnT</i>) showed upregulation at all Cu concentrations. Furthermore, the upregulation of the di-heme enzyme at concentrations above 5 µM Cu may play a crucial role in the copper switch by increasing oxygen consumption; however, the role has yet not been elucidated. We developed a quantitative assay based on the naphthalene–Molisch principle to distinguish between the sMMO- and pMMO-expressing cells, which coincided with the regulation profile of the sMMO and pMMO genes. At 0 and 3 µM Cu, the naphthol concentration was higher (8.1 and 4.2 µM, respectively) and gradually decreased to 0 µM naphthol when pMMO was expressed and acted as the sole methane oxidizer at concentrations above 5 µM Cu. Using physical protein–protein interaction, we identified seven transporters, three cell wall biosynthesis or degradation proteins, Cu resistance operon proteins, and 18 hypothetical proteins that may be involved in Cu toxicity and homeostasis. These findings shed light on the key regulatory genes of the Cu switch that will have potential implications for bioremediation and biotechnology applications.https://www.mdpi.com/2674-0389/3/1/7differential expressionmethanobactinmethanotrophpMMOprotein–protein interactionsMMO |
| spellingShingle | Dipayan Samanta Tanvi Govil Priya Saxena Lee Krumholz Venkataramana Gadhamshetty Kian Mau Goh Rajesh K. Sani Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper Methane differential expression methanobactin methanotroph pMMO protein–protein interaction sMMO |
| title | Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper |
| title_full | Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper |
| title_fullStr | Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper |
| title_full_unstemmed | Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper |
| title_short | Genetical and Biochemical Basis of Methane Monooxygenases of <i>Methylosinus trichosporium</i> OB3b in Response to Copper |
| title_sort | genetical and biochemical basis of methane monooxygenases of i methylosinus trichosporium i ob3b in response to copper |
| topic | differential expression methanobactin methanotroph pMMO protein–protein interaction sMMO |
| url | https://www.mdpi.com/2674-0389/3/1/7 |
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