Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1
The non-stereospecific α-haloalkanoic acid dehalogenase DehE from Rhizobium sp. RC1 catalyzes the removal of the halide from α-haloalkanoic acid D,L-stereoisomers and, by doing so, converts them into hydroxyalkanoic acid L,D-stereoisomers, respectively. DehE has been extensively studied to determine...
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| Format: | Article |
| Language: | English |
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Wiley-VCH Verlag
2013
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| Online Access: | http://irep.iium.edu.my/33817/1/61_fzh_JOBM_Doi_early_view.pdf |
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| author | Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Huyop, Fahrul Zaman |
| author_facet | Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Huyop, Fahrul Zaman |
| author_sort | Abdul Hamid, Azzmer Azzar |
| collection | IIUM |
| description | The non-stereospecific α-haloalkanoic acid dehalogenase DehE from Rhizobium sp. RC1 catalyzes the removal of the halide from α-haloalkanoic acid D,L-stereoisomers and, by doing so, converts them into hydroxyalkanoic acid L,D-stereoisomers, respectively. DehE has been extensively studied to determine its potential to act as a bioremediation agent, but its structure/function relationship has not been characterized. For this study, we explored the functional relevance of several putative active-site amino acids by site-specific mutagenesis. Ten active-site residues were mutated individually, and the dehalogenase activity of each of the 10 resulting mutants in soluble cell lysates against D- and L-2-chloropropionic acid was assessed. Interestingly, the mutants W34 → A, F37 → A, and S188 → A had diminished activity, suggesting that these residues are functionally relevant. Notably, the D189 → N mutant had no activity, which strongly implies that it is a catalytically important residue. Given our data, we propose a dehalogenation mechanism for DehE, which is the same as that suggested for other non-stereospecific α-haloalkanoic acid dehalogenases. To the best of our knowledge, this is the first report detailing a functional aspect for DehE, and our results could help pave the way for the bioengineering of haloalkanoic acid dehalogenases with improved catalytic properties |
| first_indexed | 2024-03-05T23:21:35Z |
| format | Article |
| id | oai:generic.eprints.org:33817 |
| institution | International Islamic University Malaysia |
| language | English |
| last_indexed | 2024-03-05T23:21:35Z |
| publishDate | 2013 |
| publisher | Wiley-VCH Verlag |
| record_format | dspace |
| spelling | oai:generic.eprints.org:338172014-11-27T08:43:53Z http://irep.iium.edu.my/33817/ Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Huyop, Fahrul Zaman Q Science (General) The non-stereospecific α-haloalkanoic acid dehalogenase DehE from Rhizobium sp. RC1 catalyzes the removal of the halide from α-haloalkanoic acid D,L-stereoisomers and, by doing so, converts them into hydroxyalkanoic acid L,D-stereoisomers, respectively. DehE has been extensively studied to determine its potential to act as a bioremediation agent, but its structure/function relationship has not been characterized. For this study, we explored the functional relevance of several putative active-site amino acids by site-specific mutagenesis. Ten active-site residues were mutated individually, and the dehalogenase activity of each of the 10 resulting mutants in soluble cell lysates against D- and L-2-chloropropionic acid was assessed. Interestingly, the mutants W34 → A, F37 → A, and S188 → A had diminished activity, suggesting that these residues are functionally relevant. Notably, the D189 → N mutant had no activity, which strongly implies that it is a catalytically important residue. Given our data, we propose a dehalogenation mechanism for DehE, which is the same as that suggested for other non-stereospecific α-haloalkanoic acid dehalogenases. To the best of our knowledge, this is the first report detailing a functional aspect for DehE, and our results could help pave the way for the bioengineering of haloalkanoic acid dehalogenases with improved catalytic properties Wiley-VCH Verlag 2013-08 Article PeerReviewed application/pdf en http://irep.iium.edu.my/33817/1/61_fzh_JOBM_Doi_early_view.pdf Abdul Hamid, Azzmer Azzar and Tengku Abdul Hamid, Tengku Haziyamin and Abdul Wahab, Roswanira and Huyop, Fahrul Zaman (2013) Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1. Journal of Basic Microbiology, 53. pp. 1-7. ISSN 0233-111X (Print) 1521-4028 (Online) http://onlinelibrary.wiley.com/doi/10.1002/jobm.201300526/abstract DOI: 10.1002/jobm.201300526 |
| spellingShingle | Q Science (General) Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Huyop, Fahrul Zaman Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title | Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title_full | Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title_fullStr | Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title_full_unstemmed | Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title_short | Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1 |
| title_sort | identification of functional residues essential for dehalogenation by the non stereospecific α haloalkanoic acid dehalogenase from rhizobium sp rc1 |
| topic | Q Science (General) |
| url | http://irep.iium.edu.my/33817/1/61_fzh_JOBM_Doi_early_view.pdf |
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