Construction of synthetic promoters for the expression of cellulases using novel cis-regulatory elements in Trichoderma reesei

• Grant number: 17/17924-1
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: December 01, 2017
• End date: April 30, 2021
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: Rafael Silva Rocha
• Grantee: Leonardo Martins Santana
• Host Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 12/22921-8 - Synthetic biology approaches for deciphering the logic of signal integration in complex bacterial promoters, AP.JP

Abstract

Microorganisms producing cellulolitic enzymes for the degradation of plant biomass have a tremendous biotechnological interest for the production of high-value compounds. In this sense, the fungus Trichoderma reesei has a notorious place as a great cellulase and hemicellulase producer and has been widely targeted for the generation of new tools and technologies to improve its capabilities. In this organism, the action of transcriptional factors controlling cellulase and hemicellulase gene expression has been widely studied. This regulation is related to the interaction of these transcriptional factors with specific regions of the target promoters, the so-called cis-regulatory elements. The project aims the construction of synthetic promoters through the computational identification and experimental validation of cis-regulatory elements related to the regulation of genes coding for biomass degradation enzymes. For this, experimental assays will be performed to analyzed the kinetics of promoters related to cellulase expression in fusion with the GFP reporter integrated into the chromosome of T. reesei. Additionally, synthetic promoters will be constructed using novel cis-regulatory elements and experimentally characterized. The objectives proposed here should contribute to increase the knowledge about the molecular mechanisms of transcriptional regulation in T. reesei. Furthermore, we expect to develop new tool for the genetic manipulation of T. reesei toward biotechnological applications. (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)

OLIVEIRA MONTEIRO, LUMMY MARIA; ARRUDA, LETICIA MAGALHAES; SANCHES-MEDEIROS, ANANDA; MARTINS-SANTANA, LEONARDO; DE FATIMA ALVES, LUANA; DEFELIPE, LUCAS; GUSTAVO TURJANSKI, ADRIAN; GUAZZARONI, MARIA-EUGENIA; DE LORENZO, VICTOR; SILVA-ROCHA, RAFAEL. Reverse Engineering of an Aspirin-Responsive Transcriptional Regulator in Escherichia coli. ACS SYNTHETIC BIOLOGY, v. 8, n. 8, p. 1890-1900, AUG 2019. (16/19179-9, 17/17924-1, 16/06323-4, 18/04810-0, 15/04309-1, 12/22921-8)

NORA, LUISA CZAMANSKI; WESTMANN, CAUA ANTUNES; MARTINS-SANTANA, LEONARDO; ALVES, LUANA DE FATIMA; OLIVEIRA MONTEIRO, LUMMY MARIA; GUAZZARONI, MARIA-EUGENIA; SILVA-ROCHA, RAFAEL. The art of vector engineering: towards the construction of next-generation genetic tools. MICROBIAL BIOTECHNOLOGY, v. 12, n. 1, p. 23-pg., 2019-01-01. (16/05472-6, 16/06323-4, 17/17924-1, 16/19179-9, 16/03763-3, 12/22921-8, 15/04309-1)

NORA, LUISA CZAMANSKI; WESTMANN, CAUA ANTUNES; MARTINS-SANTANA, LEONARDO; ALVES, LUANA DE FATIMA; OLIVEIRA MONTEIRO, LUMMY MARIA; GUAZZARONI, MARIA-EUGENIA; SILVA-ROCHA, RAFAEL. The art of vector engineering: towards the construction of next-generation genetic tools. MICROBIAL BIOTECHNOLOGY, v. 12, n. 1, p. 125-147, JAN 2019. (16/19179-9, 17/17924-1, 16/06323-4, 16/05472-6, 16/03763-3, 15/04309-1, 12/22921-8)

LEONARDO MARTINS-SANTANA; RENATO GRACIANO DE PAULA; ADRIANO GOMES SILVA; DOUGLAS CHRISTIAN BORGES LOPES; ROBERTO DO NASCIMENTO SILVA; RAFAEL SILVA-ROCHA. CRZ1 regulator and calcium cooperatively modulate holocellulases gene expression in Trichoderma reesei QM6a. GENETICS AND MOLECULAR BIOLOGY, v. 43, n. 2, 2020-05-08. (17/17924-1, 12/22921-8)

MARTINS-SANTANA, LEONARDO; NORA, LUISA C.; SANCHES-MEDEIROS, ANANDA; LOVATE, GABRIEL L.; CASSIANO, MURILO H. A.; SILVA-ROCHA, RAFAEL. Systems and Synthetic Biology Approaches to Engineer Fungi for Fine Chemical Production. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 6, OCT 3 2018. (15/22386-3, 17/17924-1, 16/11093-8, 17/04217-5, 16/03763-3, 12/22921-8)