Structural and functional characterization of the glutathione peroxidase-like thioredoxin peroxidase from the fungus Trichoderma reesei

Author(s)

Adriani, Patricia P. ^[1] ; de Paiva, Fernanda C. R. ^[2] ; de Oliveira, Gabriel S. ^[2] ; Leite, Amanda C. ^[1] ; Sanches, Adriana S. ^[1] ; Lopes, Adriana Rios ^[3] ; Dias, Marcio V. B. ^{[2, 4]} ; Chambergo, Felipe S. ^[1]

Affiliation

^[1] Univ Sao Paulo, Escola Artes Ciencias & Humanidades, 1000 Arlindo Bettio Ave, BR-03828000 Sao Paulo - Brazil

^[2] Univ Sao Paulo, Inst Ciencias Biomed, Dept Microbiol, 1374 Prof Lineu Prestes Ave, Sao Paulo - Brazil

^[3] Inst Butantan, Lab Biochem, Av Vital Brasil 1500, Sao Paulo - Brazil

^[4] Univ Warwick, Dept Chem, Gibbet Hill, Coventry CV4 7AL, W Midlands - England

• Document type: Journal article
• Source: International Journal of Biological Macromolecules; v. 167, p. 93-100, JAN 15 2021.
• Web of Science Citations: 0

Abstract

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 K-mapp = 11.7 mM, V-maxapp = 10.9 IU/mu g TrGPx, k(cat) = 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 degrees 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 antioxidantmechanism in fungi and provide information for using antioxidant enzymes in biotechnological applications. (C) 2020 Published by Elsevier B.V. (AU)

• FAPESP's process: 18/00351-1 - Applied structural biology involved in the biosynthesis of natural products: biotechnolgical aplications and study of unusual molecular reactions
• Grantee: Marcio Vinicius Bertacine Dias
• Support Opportunities: Regular Research Grants
• FAPESP's process: 17/25705-8 - Characterization and study of Enzymes to biomass degradation.
• Grantee: Felipe Santiago Chambergo Alcalde
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/05227-0 - Study and characterization of enzymes catalase and gluthathione peroxidase from Trichoderma reesei
• Grantee: Amanda Cristina Esteves Leite
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 14/24107-1 - Characterization and study of oxidases enzymes from filamentous fungi
• Grantee: Felipe Santiago Chambergo Alcalde
• Support Opportunities: Program for Research on Bioenergy (BIOEN) - Regular Program Grants