Modifying the catalytic preference of alpha-amylase toward n-alkanes for bioremediation purposes using in silico strategies
Modifying the catalytic preference of alpha-amylase toward n-alkanes for bioremediation purposes using in silico strategies
Since the beginning of oil exploration, whole ecosystems have been affected by accidents and bad practices involving petroleum compounds. In this sense, bioremediation stands out as the cheapest and most eco-friendly alternatives to reverse the damage done in oil-impacted areas. However, more efforts must be made to engineer enzymes that could be used in the bioremediation process. Interestingly, a recent work described that α-amylase, one of the most evolutionary conserved enzymes, was able to promiscuously degrade n-alkanes, a class of molecules abundant in the petroleum admixture. Considering that α-amylase is expressed in almost all known organisms, and employed in numerous biotechnological processes, using it can be a great leap toward more efficient applications of enzyme or microorganism-consortia bioremediation approaches. In this work, we employed a strict computational approach to design new α-amylase mutants with potentially enhanced catalytic efficiency toward n-alkanes. Using in silico techniques, such as molecular docking, molecular dynamics, metadynamics, and residue–residue interaction networks, we generated mutants potentially more efficient for degrading n-alkanes, L183Y, and N314A. Our results indicate that the new mutants have an increased binding rate for tetradecane, the longest n-alkane previously tested, which can reside in the catalytic center for more extended periods. Additionally, molecular dynamics and network analysis showed that the new mutations have no negative impact on protein structure than the WT. Our results aid in solidifying this enzyme as one more tool in the petroleum bioremediation toolbox.
alpha-amylase, bioremediation, metadynamics, molecular dynamics, n-alkanes, residue–residue interaction networks
Pinto, Éderson Sales Moreira
712ecaa1-8a6d-4eec-b394-4456e065edfa
Feltes, Bruno César
73ad019c-24a8-40c4-b29f-492c975159ab
Pedebos, Conrado
87801080-118f-4814-8f86-3524184b0d88
Dorn, Márcio
9d421aea-9e2b-4ab2-a9ac-b85f460bf073
15 August 2021
Pinto, Éderson Sales Moreira
712ecaa1-8a6d-4eec-b394-4456e065edfa
Feltes, Bruno César
73ad019c-24a8-40c4-b29f-492c975159ab
Pedebos, Conrado
87801080-118f-4814-8f86-3524184b0d88
Dorn, Márcio
9d421aea-9e2b-4ab2-a9ac-b85f460bf073
Pinto, Éderson Sales Moreira, Feltes, Bruno César, Pedebos, Conrado and Dorn, Márcio
(2021)
Modifying the catalytic preference of alpha-amylase toward n-alkanes for bioremediation purposes using in silico strategies.
Journal of Computational Chemistry.
(doi:10.1002/jcc.26562).
Abstract
Since the beginning of oil exploration, whole ecosystems have been affected by accidents and bad practices involving petroleum compounds. In this sense, bioremediation stands out as the cheapest and most eco-friendly alternatives to reverse the damage done in oil-impacted areas. However, more efforts must be made to engineer enzymes that could be used in the bioremediation process. Interestingly, a recent work described that α-amylase, one of the most evolutionary conserved enzymes, was able to promiscuously degrade n-alkanes, a class of molecules abundant in the petroleum admixture. Considering that α-amylase is expressed in almost all known organisms, and employed in numerous biotechnological processes, using it can be a great leap toward more efficient applications of enzyme or microorganism-consortia bioremediation approaches. In this work, we employed a strict computational approach to design new α-amylase mutants with potentially enhanced catalytic efficiency toward n-alkanes. Using in silico techniques, such as molecular docking, molecular dynamics, metadynamics, and residue–residue interaction networks, we generated mutants potentially more efficient for degrading n-alkanes, L183Y, and N314A. Our results indicate that the new mutants have an increased binding rate for tetradecane, the longest n-alkane previously tested, which can reside in the catalytic center for more extended periods. Additionally, molecular dynamics and network analysis showed that the new mutations have no negative impact on protein structure than the WT. Our results aid in solidifying this enzyme as one more tool in the petroleum bioremediation toolbox.
Text
JCC_Modifying_the_catalytic_preference_of_alpha_amylase
- Accepted Manuscript
More information
Accepted/In Press date: 4 May 2021
e-pub ahead of print date: 20 May 2021
Published date: 15 August 2021
Additional Information:
Funding Information:
Conselho Nacional de Desenvolvimento Científico e Tecnológico, Grant/Award Number: 311611 / 2018‐4; Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Grant/Award Numbers: 88881.198766/2018‐0, Finance Code 001; Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul, Grant/Award Number: 19/2551‐0001906‐8 Funding information
Funding Information:
This work was supported by grants from the Funda??o de Amparo ? Pesquisa do Estado do Rio Grande do Sul (FAPERGS) (19/2551-0001906-8), Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) (311611/2018-4), and was financed, in part, by the Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES) (DAAD/CAPES PROBRAL 88881.198766/2018-0; Finance Code 001) - Brazil.
Funding Information:
This work was supported by grants from the Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) (19/2551‐0001906‐8), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (311611/2018‐4), and was financed, in part, by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (DAAD/CAPES PROBRAL 88881.198766/2018‐0; Finance Code 001) ‐ Brazil.
Publisher Copyright:
© 2021 Wiley Periodicals LLC.
Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.
Keywords:
alpha-amylase, bioremediation, metadynamics, molecular dynamics, n-alkanes, residue–residue interaction networks
Identifiers
Local EPrints ID: 450201
URI: http://eprints.soton.ac.uk/id/eprint/450201
ISSN: 0192-8651
PURE UUID: 334b2e2a-bae2-401e-89ba-619a46776fb7
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Date deposited: 15 Jul 2021 16:40
Last modified: 06 Jun 2024 04:19
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Contributors
Author:
Éderson Sales Moreira Pinto
Author:
Bruno César Feltes
Author:
Conrado Pedebos
Author:
Márcio Dorn
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