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(Referência obtida automaticamente do Web of Science, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

Transition Path Sampling Study of the Feruloyl Esterase Mechanism

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Autor(es):
Silveira, Rodrigo L. [1, 2, 3, 4] ; Knott, Brandon C. [3] ; Pereira, Caroline S. [1, 2, 3] ; Crowley, Michael F. [3] ; Skaf, Munir S. [1, 2] ; Beckham, Gregg T. [3]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Estadual Campinas, Inst Chem, BR-13084862 Campinas, SP - Brazil
[2] Univ Estadual Campinas, Ctr Comp Engn & Sci, BR-13084862 Campinas, SP - Brazil
[3] Natl Renewable Energy Lab, Renewable Resources & Enabling Sci Ctr, Golden, CO 80401 - USA
[4] Univ Fed Rio de Janeiro, Inst Chem, BR-21941909 Rio De Janeiro, RJ - Brazil
Número total de Afiliações: 4
Tipo de documento: Artigo Científico
Fonte: Journal of Physical Chemistry B; v. 125, n. 8, p. 2018-2030, MAR 4 2021.
Citações Web of Science: 0
Resumo

Serine hydrolases cleave peptide and ester bonds and are ubiquitous in nature, with applications in biotechnology, in materials, and as drug targets. The serine hydrolase two-step mechanism employs a serine-histidine-aspartate/glutamate catalytic triad, where the histidine residue acts as a base to activate poor nucleophiles (a serine residue or a water molecule) and as an acid to allow the dissociation of poor leaving groups. This mechanism has been the subject of debate regarding how histidine shuttles the proton from the nucleophile to the leaving group. To elucidate the reaction mechanism of serine hydrolases, we employ quantum mechanics/molecular mechanics-based transition path sampling to obtain the reaction coordinate using the Aspergillus niger feruloyl esterase A (AnFaeA) as a model enzyme. The optimal reaction coordinates include terms involving nucleophilic attack on the carbonyl carbon and proton transfer to, and dissociation of, the leaving group. During the reaction, the histidine residue undergoes a reorientation on the time scale of hundreds of femtoseconds that supports the ``moving histidine{''} mechanism, thus calling into question the ``ring flip{''} mechanism. We find a concerted mechanism, where the transition state coincides with the tetrahedral intermediate with the histidine residue pointed between the nucleophile and the leaving group. Moreover, motions of the catalytic aspartate toward the histidine occur concertedly with proton abstraction by the catalytic histidine and help stabilize the transition state, thus partially explaining how serine hydrolases enable poor nucleophiles to attack the substrate carbonyl carbon. Rate calculations indicate that the second step (deacylation) is rate-determining, with a calculated rate constant of 66 s(-1). Overall, these results reveal the pivotal role of active-site dynamics in the catalytic mechanism of AnFaeA, which is likely similar in other serine hydrolases. (AU)

Processo FAPESP: 15/25031-1 - Dinâmica molecular de enzimas modificadores de de carbohidratos para desconstrução e valorização de biomassa lignocelulósica
Beneficiário:Caroline Simões Pereira
Linha de fomento: Bolsas no Brasil - Doutorado
Processo FAPESP: 17/01151-3 - Simulações de QM/MM-TPS de transglicosilases líticas
Beneficiário:Caroline Simões Pereira
Linha de fomento: Bolsas no Exterior - Estágio de Pesquisa - Doutorado
Processo FAPESP: 16/22956-7 - Simulações de QM/MM híbridas de feruloil esterases: mecanismo de clivagem de complexos lignina-carboidrato em paredes celulares de plantas
Beneficiário:Rodrigo Leandro Silveira
Linha de fomento: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado
Processo FAPESP: 13/08293-7 - CECC - Centro de Engenharia e Ciências Computacionais
Beneficiário:Munir Salomao Skaf
Linha de fomento: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
Processo FAPESP: 14/10448-1 - Aspectos moleculares da arquitetura de paredes celulares de plantas
Beneficiário:Rodrigo Leandro Silveira
Linha de fomento: Bolsas no Brasil - Pós-Doutorado