Busca avançada
Ano de início
Entree
(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.)

Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization

Texto completo
Autor(es):
de Oliveira, Gabriel Stephani [1] ; Adriani, Patricia Pereira [1] ; Borges, Flavia Garcia [1] ; Lopes, Adriana Rios [2] ; Campana, Patricia T. [1] ; Chambergo, Felipe S. [1]
Número total de Autores: 6
Afiliação do(s) autor(es):
[1] Univ Sao Paulo, Escola Artes Ciencias & Humanidades, Av Arlindo Bettio 1000, Sao Paulo - Brazil
[2] Inst Butantan, Av Vital Brasil 1500, Sao Paulo - Brazil
Número total de Afiliações: 2
Tipo de documento: Artigo Científico
Fonte: International Journal of Biological Macromolecules; v. 89, p. 569-574, AUG 2016.
Citações Web of Science: 6
Resumo

Epoxide hydrolases (EHs) are enzymes that are present in all living organisms and catalyze the hydrolysis of epoxides to the corresponding vicinal diols. EHs have biotechnological potential in chiral chemistry. We report the cloning, purification, enzymatic activity, and conformational analysis of the TrEH gene from Trichoderma reesei strain QM9414 using circular dichroism spectroscopy. The EH gene has an open reading frame encoding a protein of 343 amino acid residues, resulting in a molecular mass of 38.2 kDa. The enzyme presents an optimum pH of 7.2, and it is highly active at temperatures ranging from 23 to 50 degrees C and thermally inactivated at 70 degrees C (t(1/2) = 7.4 min). The Michaelis constants (K-m) were 4.6 mM for racemic substrate, 21.7 mM for (R)-(+)-styrene oxide and 3.0mM for (S)-()-styrene oxide. The k(cat)/K-m analysis indicated that TrEH is enantioselective and preferentially hydrolyzes (S)-(-)-styrene oxide. The conformational stability studies suggested that, despite the extreme conditions (high temperatures and extremely acid and basic pHs), TrEH is able to maintain a considerable part of its regular structures, including the preservation of the native cores in some cases. The recombinant protein showed enantioselectivity that was distinct from other fungus EHs, making this protein a potential biotechnological tool. (C) 2016 Elsevier B.V. All rights reserved. (AU)

Processo FAPESP: 15/03329-9 - Clonagem e caracterização de enzimas epóxido hidrolases de Trichoderma reesei
Beneficiário:Gabriel Stephani de Oliveira
Linha de fomento: Bolsas no Brasil - Doutorado Direto
Processo FAPESP: 14/24107-1 - Estudo e caracterização de enzimas oxidases produzidas por fungos filamentosos
Beneficiário:Felipe Santiago Chambergo Alcalde
Linha de fomento: Auxílio à Pesquisa - Programa BIOEN - Regular