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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 halogenated 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 muta...

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Autores principales: Abdul Hamid, Azzmer Azzar, Tengku Abdul Hamid, Tengku Haziyamin, Abdul Wahab, Roswanira, Omar, Mohd. Shahir Shamsir, Huyop, Fahrul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376737/
https://www.ncbi.nlm.nih.gov/pubmed/25816329
http://dx.doi.org/10.1371/journal.pone.0121687
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author Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
author_facet Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
author_sort Abdul Hamid, Azzmer Azzar
collection PubMed
description The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated 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 substrate 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.
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spelling pubmed-43767372015-04-04 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 Omar, Mohd. Shahir Shamsir Huyop, Fahrul PLoS One Research Article The non-stereospecific α-haloalkanoic acid dehalogenase E (DehE) degrades many halogenated 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 substrate 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. Public Library of Science 2015-03-27 /pmc/articles/PMC4376737/ /pubmed/25816329 http://dx.doi.org/10.1371/journal.pone.0121687 Text en © 2015 Abdul Hamid et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Abdul Hamid, Azzmer Azzar
Tengku Abdul Hamid, Tengku Haziyamin
Abdul Wahab, Roswanira
Omar, Mohd. Shahir Shamsir
Huyop, Fahrul
An S188V Mutation Alters Substrate Specificity of Non-Stereospecific α-Haloalkanoic Acid Dehalogenase E (DehE)
title 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_short 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)
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4376737/
https://www.ncbi.nlm.nih.gov/pubmed/25816329
http://dx.doi.org/10.1371/journal.pone.0121687
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