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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Main Authors:	Abdul Hamid, Azzmer Azzar, Tengku Abdul Hamid, Tengku Haziyamin, Abdul Wahab, Roswanira, Huyop, Fahrul Zaman
Format:	Article
Language:	English
Published:	Wiley-VCH Verlag 2013
Subjects:	Q Science (General)
Online Access:	http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/1/61_fzh_JOBM_Doi_early_view.pdf

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spelling	iium-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
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	Q Science (General)
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
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
format	Article
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
title	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_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_sort	identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from rhizobium sp. rc1
publisher	Wiley-VCH Verlag
publishDate	2013
url	http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/ http://irep.iium.edu.my/33817/1/61_fzh_JOBM_Doi_early_view.pdf
first_indexed	2023-09-18T20:48:55Z
last_indexed	2023-09-18T20:48:55Z
_version_	1777409861967937536

Identification of functional residues essential for dehalogenation by the non-stereospecific α-haloalkanoic acid dehalogenase from Rhizobium sp. RC1

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