Cloning and characterization of the enzymes epoxide hydrolases from Trichoderma reesei

• Grant number: 15/03329-9
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: October 01, 2015
• End date: March 25, 2018
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Felipe Santiago Chambergo Alcalde
• Grantee: Gabriel Stephani de Oliveira
• Host Institution: Escola de Artes, Ciências e Humanidades (EACH). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 16/12859-4 - Screening of Substrates and Inhibitors for epoxide hydrolase from Trichoderma reesei, BE.EP.DD

Abstract

Enzymes are important biological molecules widely studied and used in economic activities. Epoxide hydrolase enzymes catalyze the hydrolysis of epoxides to their corresponding diols, and several studies have shown its great potential for biotechnological applications in industrial processes and bioremediation. This project describes the steps already completed in the master's program, as the identification and characterization of two genes encoding epoxide hydrolases enzymes (TrEH 1 and TrEH 2) of the fungus Trichoderma reesei and cloning in pProEX-HTa expression vector (Life Technologies, USA), expression in Escherichia coli BL 21, and purifying of the epoxide hydrolase 2 (TrEH 2). Developed preliminary tests show enzymatic activity and enantioselectivity on the substrate styrene oxide (epoxide). For the continuation of the project (direct doctorate) proposes the expression and purification of recombinant protein TrEH 1 and with the two enzymes (TrEH 1 and TrEH 2), we intend to conduct the kinetic enzymatic characterization, tests of enantioselectivity determination with High Performance Liquid Chromatography (HPLC) and structural predictions. This proposal is the first in the description and characterization of the Trichoderma reesei's epoxide hydrolases. (AU)

Scientific publications

(5)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE OLIVEIRA, GABRIEL S.; ADRIANI, PATRICIA P.; WU, HAO; MORISSEAU, CHRISTOPHE; HAMMOCK, BRUCE D.; CHAMBERGO, FELIPE S.. Substrate and inhibitor selectivity, and biological activity of an epoxide hydrolase from Trichoderma reesei. MOLECULAR BIOLOGY REPORTS, v. 46, n. 1, p. 371-379, FEB 2019. (16/12859-4, 15/03329-9, 14/24107-1, 17/25705-8)

DEOLIVEIRA, GABRIEL STEPHANI; ADRIANI, PATRICIA PEREIRA; BORGES, FLAVIA GARCIA; LOPES, ADRIANA RIOS; CAMPANA, PATRICIA T.; CHAMBERGO, FELIPE S.. Data set of optimal parameters for colorimetric red assay of epoxide hydrolase activity. DATA IN BRIEF, v. 8, p. 436-440, SEP 2016. (14/24107-1, 15/03329-9)

WILSON, CAROLINA; DE OLIVEIRA, GABRIEL S.; ADRIANI, PATRICIA P.; CHAMBERGO, FELIPE S.; DIAS, MARCIO V. B.. Structure of a soluble epoxide hydrolase identified in Trichoderma reesei. BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, v. 1865, n. 8, p. 1039-1045, AUG 2017. (15/09188-8, 10/15971-3, 14/24107-1, 15/03329-9)

DE OLIVEIRA, GABRIEL STEPHANI; ADRIANI, PATRICIA PEREIRA; BORGES, FLAVIA GARCIA; LOPES, ADRIANA RIOS; CAMPANA, PATRICIA T.; CHAMBERGO, FELIPE S.. Epoxide hydrolase of Trichoderma reesei: Biochemical properties and conformational characterization. International Journal of Biological Macromolecules, v. 89, p. 569-574, AUG 2016. (14/24107-1, 15/03329-9)

DE OLIVEIRA, GABRIEL S.; ADRIANI, PATRICIA P.; RIBEIRO, JOAO AUGUSTO; MORISSEAU, CHRISTOPHE; HAMMOCK, BRUCE D.; DIAS, MARCIO VINICIUS B.; CHAMBERGO, FELIPE S.. The molecular structure of an epoxide hydrolase from Trichoderma reesei in complex with urea or amide-based inhibitors. International Journal of Biological Macromolecules, v. 129, p. 653-658, MAY 15 2019. (13/15906-5, 16/12859-4, 15/03329-9, 15/09188-8, 18/00351-1, 14/24107-1, 17/25705-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

OLIVEIRA, Gabriel Stephani de. Cloning and characterization of the enzyme epoxide hydrolase of the Trichoderma reesei.. 2018. Doctoral Thesis - Universidade de São Paulo (USP). Instituto de Ciências Biomédicas (ICB/SDI) São Paulo.