Developing an E. coli heterologous expression system for characterizing a marine debrominase from Roseobacter sp.
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Le résumé fourni par la source
ABSTRACT Reductive dehalogenases (RDases) are a family of enzymes that employ a cobamide cofactor and two iron-sulfur clusters to selectively cleave carbon-halogen bonds. The identification of yet-uncharacterized RDases is crucial for advancing our understanding of dehalogenation processes in both natural and engineered environments, particularly for bioremediation. In this study, we developed a heterologous expression system in Escherichia coli DH5α to produce a chloroform-reducing RDase (TmrA) from Dehalobacter sp. UNSWDHB, using a pTrcHisA vector without requiring an additional vector for vitamin B 12 transporters. By co-expressing the pRSFDuet-BtuCEDFB plasmid, we further enhanced active TmrA production without relying on the T7 promoter system. Using this system, we achieved the first heterologous expression of an RDase (the debrominase Ros_A3X954) from a marine heterotrophic Roseobacter strain, without chemical cofactor reconstitution. Ros_A3X954 catalyzes conversion of 2,6-dibromophenol to 2-bromophenol with a specific activity of 21.8 ± 1.0 nmol s −1 mg −1 . The enzyme exhibited optimal activity at pH 6.0 and demonstrated significant thermal stability by maintaining full catalytic capacity across a broad temperature range (20–50°C), a key advantage for applications in thermally fluctuating marine environments. These findings expand our understanding of RDase functional diversity in common marine microbiota and highlight the significant application potential of Roseobacter for bioremediation of bromophenol pollution. IMPORTANCE Organohalide-respiring bacteria (OHRB) play a crucial role in bioremediation by degrading halogenated pollutants using cobamide-dependent reductive dehalogenases (RDases). While these enzymes hold great promise for bioremediation, their practical application has been limited by difficulties in production and heterologous expression. This study successfully developed an Escherichia coli DH5α expression system for RDases without requiring vitamin B 12 transporters expression vector, significantly simplifying production process. A bromophenol-degrading RDase (Ros_A3X954) from Roseobacter sp. was discovered, exhibiting high activity at pH 6.0 and thermal stability (20–50°C). This study not only provides a scalable platform for RDase exploration but also highlights Roseobacter ’s ecological role in bromine cycling, advancing bioremediation strategies for persistent pollutants.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé, mais le titre doit être comparé manuellement.
- Titre Crossref
- Developing an <i>E. coli</i> heterologous expression system for characterizing a marine debrominase from <i>Roseobacter</i> sp.
- Date Crossref
- 22/04/2026
- Éditeur
- American Society for Microbiology
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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South China Normal University pays non établi dans la noticeUniversité ou école supérieure
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Guangzhou Institute of Geochemistry pays non établi dans la noticeStructure de recherche
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University of Chinese Academy of Sciences pays non établi dans la noticeUniversité ou école supérieure
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Guangdong-Hongkong-Macau Joint Laboratory of Collaborative Innovation for Environmental Quality pays non établi dans la noticeUniversité ou école supérieure
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Helmholtz Centre for Environmental Research pays non établi dans la noticeStructure de recherche
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Technische Universität Berlin pays non établi dans la noticeUniversité ou école supérieure
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School of Life Sciences Guangdong Provincial Key Laboratory of Biotechnology for Plant Development pays non établi dans la noticeUniversité ou école supérieure
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State Key Laboratory of Advanced Environmental Technology pays non établi dans la noticeStructure de recherche
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Ltd Shenzhen Readline Biotech CO. pays non établi dans la noticeEntreprise
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Guangdong Key Laboratory of Environmental Protection and Resources and Utilization pays non établi dans la noticeStructure de recherche
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Guangdong-Hong Kong-Maco Joint Laboratory for Environmental Pollution and Control pays non établi dans la noticeStructure de recherche
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Chair of Geobiotechnology pays non établi dans la noticeInstitution
South China Normal University, Guangzhou Institute of Geochemistry et University of Chinese Academy of Sciences, avec 9 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.