An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)

An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)

The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halo- genated compounds but is ineffective against β-halogenated compounds such as 3- chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing th...

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Main Authors: Abdul Hamid, Azzmer Azzar, Tengku Abdul Hamid, Tengku Haziyamin, Abdul Wahab, Roswanira, Shamsir, Mohd Shahir, Huyop, Fahrul Zaman
Format: Article
Language:English
Published: The Public Library of Science (PLoS) 2015
Subjects:
QR Microbiology
TP248.13 Biotechnology
Online Access:http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/1/42599.pdf
id iium-42599
recordtype eprints
spelling iium-425992017-11-03T02:31:34Z http://irep.iium.edu.my/42599/ An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) Abdul Hamid, Azzmer Azzar Tengku Abdul Hamid, Tengku Haziyamin Abdul Wahab, Roswanira Shamsir, Mohd Shahir Huyop, Fahrul Zaman QR Microbiology TP248.13 Biotechnology The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halo- genated compounds but is ineffective against β-halogenated compounds such as 3- chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves sub- strate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2- chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by K m . This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies. The Public Library of Science (PLoS) 2015-03-27 Article PeerReviewed application/pdf en http://irep.iium.edu.my/42599/1/42599.pdf Abdul Hamid, Azzmer Azzar and Tengku Abdul Hamid, Tengku Haziyamin and Abdul Wahab, Roswanira and Shamsir, Mohd Shahir and Huyop, Fahrul Zaman (2015) An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE). PLoS ONE, 10 (3). pp. 1-21. ISSN 1932-6203 http://www.plosone.org/ :10.1371/journal.pone.0121687
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
topic QR Microbiology
TP248.13 Biotechnology
spellingShingle QR Microbiology
TP248.13 Biotechnology
Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Shamsir, Mohd Shahir
Huyop, Fahrul Zaman
An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
description The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halo- genated compounds but is ineffective against β-halogenated compounds such as 3- chloropropionic acid (3CP). Using molecular dynamics (MD) simulations and site-directed mutagenesis we show here that introducing the mutation S188V into DehE improves sub- strate specificity towards 3CP. MD simulations showed that residues W34, F37, and S188 of DehE were crucial for substrate binding. DehE showed strong binding ability for D-2- chloropropionic acid (D-2CP) and L-2-chloropropionic acid (L-2CP) but less affinity for 3CP. This reduced affinity was attributed to weak hydrogen bonding between 3CP and residue S188, as the carboxylate of 3CP forms rapidly interconverting hydrogen bonds with the backbone amide and side chain hydroxyl group of S188. By replacing S188 with a valine residue, we reduced the inter-molecular distance and stabilised bonding of the carboxylate of 3CP to hydrogens of the substrate-binding residues. Therefore, the S188V can act on 3CP, although its affinity is less strong than for D-2CP and L-2CP as assessed by K m . This successful alteration of DehE substrate specificity may promote the application of protein engineering strategies to other dehalogenases, thereby generating valuable tools for future bioremediation technologies.
format Article
author Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Shamsir, Mohd Shahir
Huyop, Fahrul Zaman
author_facet Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Shamsir, Mohd Shahir
Huyop, Fahrul Zaman
author_sort Abdul Hamid, Azzmer Azzar
title An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
title_short An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
title_full An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
title_fullStr An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
title_full_unstemmed An S188V mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase E (DehE)
title_sort s188v mutation alters substrate specificity of non-stereospecific α-haloalkanoic acid dehalogenase e (dehe)
publisher The Public Library of Science (PLoS)
publishDate 2015
url http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/
http://irep.iium.edu.my/42599/1/42599.pdf
first_indexed 2023-09-18T21:00:41Z
last_indexed 2023-09-18T21:00:41Z
_version_ 1777410602728161280

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