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Structural and functional characterization of the glutathione peroxidase-like thioredoxin peroxidase from the fungus Trichoderma reesei.
Adriani, Patricia P; de Paiva, Fernanda C R; de Oliveira, Gabriel S; Leite, Amanda C; Sanches, Adriana S; Lopes, Adriana Rios; Dias, Marcio V B; Chambergo, Felipe S.
Afiliação
  • Adriani PP; Escola de Artes, Ciências e Humanidades, Universidade de São Paulo, 1000 Arlindo Bettio Avenue, São Paulo, Brazil.
  • de Paiva FCR; Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, 1374 Professor Lineu Prestes Avenue, São Paulo, Brazil.
  • de Oliveira GS; Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, 1374 Professor Lineu Prestes Avenue, São Paulo, Brazil.
  • Leite AC; Escola de Artes, Ciências e Humanidades, Universidade de São Paulo, 1000 Arlindo Bettio Avenue, São Paulo, Brazil.
  • Sanches AS; Escola de Artes, Ciências e Humanidades, Universidade de São Paulo, 1000 Arlindo Bettio Avenue, São Paulo, Brazil.
  • Lopes AR; Laboratory of Biochemistry, Instituto Butantan, Av. Vital Brasil, 1500 São Paulo, Brazil.
  • Dias MVB; Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, 1374 Professor Lineu Prestes Avenue, São Paulo, Brazil; Department of Chemistry, University of Warwick, Gibbet Hill, Coventry CV4 7AL, United Kingdom of Great Britain and Northern Ireland.
  • Chambergo FS; Escola de Artes, Ciências e Humanidades, Universidade de São Paulo, 1000 Arlindo Bettio Avenue, São Paulo, Brazil. Electronic address: fscha@usp.br.
Int J Biol Macromol ; 167: 93-100, 2021 Jan 15.
Article em En | MEDLINE | ID: mdl-33259843
Glutathione peroxidases (GPx) are a family of enzymes with the ability to reduce organic and inorganic hydroperoxides to the corresponding alcohols using glutathione or thioredoxin as an electron donor. Here, we report the functional and structural characterization of a GPx identified in Trichoderma reesei (TrGPx). TrGPx was recombinantly expressed in a bacterial host and purified using affinity. Using a thioredoxin coupled assay, TrGPx exhibited activity of 28 U and 12.5 U in the presence of the substrates H2O2 and t-BOOH, respectively, and no activity was observed when glutathione was used. These results indicated that TrGPx is a thioredoxin peroxidase and hydrolyses H2O2 better than t-BOOH. TrGPx kinetic parameters using a pyrogallol assay resulted at Kmapp = 11.7 mM, Vmaxapp = 10.9 IU/µg TrGPx, kcat = 19 s-1 and a catalytic efficiency of 1.6 mM-1 s-1 to H2O2 as substrate. Besides that, TrGPx demonstrated an optimum pH ranging from 9.0-12.0 and a half-life of 36 min at 80 °C. TrGPx 3D-structure was obtained in a reduced state and non-catalytic conformation. The overall fold is similar to the other phospholipid-hydroperoxide glutathione peroxidases. These data contribute to understand the antioxidant mechanism in fungi and provide information for using antioxidant enzymes in biotechnological applications.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Peroxirredoxinas / Hypocreales Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Peroxirredoxinas / Hypocreales Idioma: En Revista: Int J Biol Macromol Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Holanda