Development and/or improvement of enzymatic cocktails designed for plant biomass hydrolysis: use of site-directed mutagenesis and chimeric proteins design

• Grant number: 16/19775-0
• Support Opportunities: Scholarships in Brazil - Post-Doctorate
• Effective date (Start): March 01, 2017
• Effective date (End): October 13, 2019
• Field of knowledge: Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
• Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: Anete Pereira de Souza
• Grantee: Clelton Aparecido dos Santos
• Host Institution: Centro de Biologia Molecular e Engenharia Genética (CBMEG). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated scholarship(s): 17/17390-7 - Substrate Specificity, Unusual Enzymatic Activity, and Synergistic Relationships of beta-glucosidases from Trichoderma harzianum, BE.EP.PD

Abstract

The conversion of sugar derived from plant biomass degradation via enzymatic hydrolysis for biofuels production is a challenge. Therefore, the search for new microorganisms and enzymes that can increase the efficiency of the saccharification of cellulosic substrates remains an important and high-priority area of study. Using a rational approach, comparing RNA-Seq and structural genomics data, our group has been able to identify the major beta-glucosidase expressed by Trichoderma harzianum (ThBgl) under biomass degradation conditions. However, after the structural and functional ThBgl characterization, the results showed that although this enzyme has been produced in high yield and purity, it did not have the two major properties desired for its effective use to supplement enzyme cocktails to the degradation of biomass - thermostability, and highglucose tolerance. In this context, the goals of this project are to provide new insights into protein engineering field, especially those related in the degradation of plant biomass, with the implementation of a platform using site-directed mutagenesis (SDM) and chimeric proteins design for the development of more efficient enzymes to be used in the supplemental enzyme cocktails to the saccharification of cellulosic substrates. (AU)

Scientific publications

(4)

(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)

FERREIRA FILHO, JAIRE ALVES; CRIVELENTE HORTA, MARIA AUGUSTA; BELOTI, LILIAN LUZIA; DOS SANTOS, CLELTON APARECIDO; DE SOUZA, ANETE PEREIRA. Carbohydrate-active enzymes in Trichoderma harzianum: a bioinformatic analysis bioprospecting for key enzymes for the biofuels industry. BMC Genomics, v. 18, OCT 12 2017. (16/19775-0, 15/09202-0, 14/18856-1)

LEAL MOTTA, MARIA LORENZA; FERREIRA FILHO, JAIRE ALVES; DE MELO, RICARDO RODRIGUES; ZANPHORLIN, LETICIA MARIA; DOS SANTOS, CLELTON APARECIDO; DE SOUZA, ANETE PEREIRA. A novel fungal metal-dependent alpha-l-arabinofuranosidase of family 54 glycoside hydrolase shows expanded substrate specificity. SCIENTIFIC REPORTS, v. 11, n. 1, MAY 26 2021. (17/14253-9, 15/09202-0, 18/19660-4, 16/19775-0, 19/08855-1)

CRIVELENTE HORTA, MARIA AUGUSTA; FERREIRA FILHO, JAIRE ALVES; MURAD, NATALIA FARAJ; SANTOS, EIDY DE OLIVEIRA; DOS SANTOS, CLELTON APARECIDO; MENDES, JULIANO SALES; BRANDAO, MARCELO MENDES; AZZONI, SINDELIA FREITAS; DE SOUZA, ANETE PEREIRA. Network of proteins, enzymes and genes linked to biomass degradation shared by Trichoderma species. SCIENTIFIC REPORTS, v. 8, JAN 22 2018. (11/00417-3, 14/18856-1, 15/09202-0, 15/50612-8, 16/19775-0)

FERREIRA FILHO, JAIRE A.; HORTA, MARIA AUGUSTA C.; DOS SANTOS, CLELTON A.; ALMEIDA, DEBORAH A.; MURAD, NATALIA F.; MENDES, JULIANO S.; SFORCA, DANILO A.; SILVA, CLAUDIO BENICIO C.; CRUCELLO, ALINE; DE SOUZA, ANETE P.. ``Integrative genomic analysis of the bioprospection of regulators and accessory enzymes associated with cellulose degradation in a filamentous fungus (Trichoderma harzianum){''}. BMC Genomics, v. 21, n. 1, NOV 2 2020. (17/26781-0, 17/17782-2, 15/09202-0, 18/19660-4, 16/19775-0, 14/18856-1)