Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC
Abstract Malachite green (MG) is used as a dye for materials such as wood, cotton, and nylon, and is used in aquaculture to prevent fungal and protozoan diseases. However, it is highly toxic, with carcinogenic, mutagenic, and teratogenic properties, resulting in bans worldwide. Despite this, MG is s...
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Nature Portfolio
2025-01-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-024-84609-4 |
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| author | Shanza Bibi Callum W. Breeze Vusqa Jadoon Anum Fareed Alina Syed Rebecca L. Frkic Habiba Zaffar Muhammad Ali Iftikhar Zeb Colin J. Jackson Tatheer Alam Naqvi |
| author_facet | Shanza Bibi Callum W. Breeze Vusqa Jadoon Anum Fareed Alina Syed Rebecca L. Frkic Habiba Zaffar Muhammad Ali Iftikhar Zeb Colin J. Jackson Tatheer Alam Naqvi |
| author_sort | Shanza Bibi |
| collection | DOAJ |
| description | Abstract Malachite green (MG) is used as a dye for materials such as wood, cotton, and nylon, and is used in aquaculture to prevent fungal and protozoan diseases. However, it is highly toxic, with carcinogenic, mutagenic, and teratogenic properties, resulting in bans worldwide. Despite this, MG is still frequently used in many countries due to its efficacy and economy. MG is persistent in the environment and so requires degradative intervention. In this work we isolated Bacillus pacificus ROC1 strain from a salt flat in Pakistan that had the ability to aerobically detoxify MG, as determined by bacterio- and phyto-toxicity assays. We demonstrate immobilized B. pacificus ROC1 can effectively detoxify MG, which highlights a potential method for its biodegradation. Genomic sequencing identified three candidate azo-reductases within B. pacificus ROC1 that could be responsible for the MG-degrading activity. These were cloned, expressed and purified from Escherichia coli, with one (AzrC), catalyzing the reduction of MG to leuco-MG in vitro. AzrC was crystallised and MG was captured within the active site in a Michaelis complex, providing structural insight into the reduction mechanism. Altogether, this work identifies a bacterium capable of aerobically degrading a major industrial pollutant and characterizes the molecular basis for this activity. |
| format | Article |
| id | doaj-art-df79205c59c94576b3fd465bd3bd3532 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-df79205c59c94576b3fd465bd3bd35322025-02-02T12:15:50ZengNature PortfolioScientific Reports2045-23222025-01-0115111510.1038/s41598-024-84609-4Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrCShanza Bibi0Callum W. Breeze1Vusqa Jadoon2Anum Fareed3Alina Syed4Rebecca L. Frkic5Habiba Zaffar6Muhammad Ali7Iftikhar Zeb8Colin J. Jackson9Tatheer Alam Naqvi10Department of Biotechnology, COMSATS University IslamabadResearch School of Chemistry, The Australian National UniversityDepartment of Biotechnology, COMSATS University IslamabadDepartment of Biotechnology, COMSATS University IslamabadDepartment of Biotechnology, COMSATS University IslamabadResearch School of Chemistry, The Australian National UniversityDepartment of Environmental Sciences, COMSATS University IslamabadDepartment of Biotechnology, COMSATS University IslamabadDepartment of Biotechnology, COMSATS University IslamabadResearch School of Chemistry, The Australian National UniversityDepartment of Biotechnology, COMSATS University IslamabadAbstract Malachite green (MG) is used as a dye for materials such as wood, cotton, and nylon, and is used in aquaculture to prevent fungal and protozoan diseases. However, it is highly toxic, with carcinogenic, mutagenic, and teratogenic properties, resulting in bans worldwide. Despite this, MG is still frequently used in many countries due to its efficacy and economy. MG is persistent in the environment and so requires degradative intervention. In this work we isolated Bacillus pacificus ROC1 strain from a salt flat in Pakistan that had the ability to aerobically detoxify MG, as determined by bacterio- and phyto-toxicity assays. We demonstrate immobilized B. pacificus ROC1 can effectively detoxify MG, which highlights a potential method for its biodegradation. Genomic sequencing identified three candidate azo-reductases within B. pacificus ROC1 that could be responsible for the MG-degrading activity. These were cloned, expressed and purified from Escherichia coli, with one (AzrC), catalyzing the reduction of MG to leuco-MG in vitro. AzrC was crystallised and MG was captured within the active site in a Michaelis complex, providing structural insight into the reduction mechanism. Altogether, this work identifies a bacterium capable of aerobically degrading a major industrial pollutant and characterizes the molecular basis for this activity.https://doi.org/10.1038/s41598-024-84609-4 |
| spellingShingle | Shanza Bibi Callum W. Breeze Vusqa Jadoon Anum Fareed Alina Syed Rebecca L. Frkic Habiba Zaffar Muhammad Ali Iftikhar Zeb Colin J. Jackson Tatheer Alam Naqvi Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC Scientific Reports |
| title | Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC |
| title_full | Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC |
| title_fullStr | Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC |
| title_full_unstemmed | Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC |
| title_short | Isolation, identification, and characterisation of the malachite green detoxifying bacterial strain Bacillus pacificus ROC1 and the azoreductase AzrC |
| title_sort | isolation identification and characterisation of the malachite green detoxifying bacterial strain bacillus pacificus roc1 and the azoreductase azrc |
| url | https://doi.org/10.1038/s41598-024-84609-4 |
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