Integrating protein engineering and synthetic biology to develop new high-throughput selection systems

• Grant number: 16/18827-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: June 01, 2017
• End date: February 29, 2020
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
• Principal Investigator: María Eugenia Guazzaroni
• Grantee: Lucas Ferreira Ribeiro
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 15/04309-1 - Novel approaches to improve functional screening of biocatalysts in metagenomic libraries, AP.JP

Abstract

The evolution of fields as protein engineering and metagenomics depends on the development of tools to exploit, with efficiency, the immense natural diversity in which the target enzymes are immersed. Therefore, to explore this astronomical sample space is necessary to create high throughput screening or selection (HTS) systems more efficient. However, despite the advancement in the construction of new strategies of HTS, many of these strategies typically have low rates of target enzymes identification. Most of the limitations encountered during the development of new HTS systems can be circumvented by synthetic biology approaches. In this project, we are proposing the development of new HTS technologies to identify enzymes for biomass degradation. These technologies will be developed from a bridge between synthetic biology and protein engineering, in order to create gene circuits capable of recognizing the product of the enzymatic activity and convert this signal on a phenotypic change. Thus, the HTS technology here proposed and named Biomass Activated Switch System (BASS) could work as a more efficient selection system helping to alleviate the limitations encountered during the process of search for new enzymes in directed evolution or metagenomics libraries. (AU)

Scientific publications

(9)

(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)

RIBEIRO, LUCAS F.; AMARELLE, VANESA; RIBEIRO, LILIANE F. C.; GUAZZARONI, MARIA-EUGENIA. Converting a Periplasmic Binding Protein into a Synthetic Biosensing Switch through Domain Insertion. BIOMED RESEARCH INTERNATIONAL, 2019. (16/20358-5, 15/04309-1, 16/18827-7)

RIBEIRO, LUCAS FERREIRA; AMARELLE, VANESA; ALVES, LUANA DE FATIMA; VIANA DE SIQUEIRA, GUILHERME MARCELINO; LOVATE, GABRIEL LENCIONI; BORELLI, TIAGO CABRAL; GUAZZARONI, MARIA-EUGENIA. Genetically Engineered Proteins to Improve Biomass Conversion: New Advances and Challenges for Tailoring Biocatalysts. Molecules, v. 24, n. 16, AUG 2019. (16/18827-7, 15/04309-1, 18/18158-3, 18/07261-8, 19/00390-0, 16/06323-4)

BORELLI, TIAGO CABRAL; LOVATE, GABRIEL LENCIONI; SCARANELLO, ANA FLAVIA TONELLI; RIBEIRO, LUCAS FERREIRA; ZARAMELA, LIVIA; PEREIRA-DOS-SANTOS, FELIPE MARCELO; SILVA-ROCHA, RAFAEL; GUAZZARONI, MARIA-EUGENIA. Combining Functional Genomics and Whole-Genome Sequencing to Detect Antibiotic Resistance Genes in Bacterial Strains Co-Occurring Simultaneously in a Brazilian Hospital. ANTIBIOTICS-BASEL, v. 10, n. 4, APR 2021. (19/00390-0, 15/04309-1, 19/06672-7, 16/18827-7, 19/15675-0, 18/18158-3)

KIKUTI MANCILIO, LUCCA BONJY; RIBEIRO, GUILHERME AUGUSTO; LOPES, ERICA MENDES; KISHI, LUCIANO TAKESHI; MARTINS-SANTANA, LEONARDO; VIANA DE SIQUEIRA, GUILHERME MARCELINO; ANDRADE, ADALGISA RODRIGUES; GUAZZARONI, MARIA-EUGENIA; REGINATTO, VALERIA. Unusual microbial community and impact of iron and sulfate on microbial fuel cell ecology and performance. CURRENT RESEARCH IN BIOTECHNOLOGY, v. 2, p. 10-pg., 2020-11-01. (18/00789-7, 18/15528-4, 15/04309-1, 16/18827-7)

RIBEIRO, LUCAS F.; RIBEIRO, LILIANE F. C.; BARRETO, MATHEUS Q.; WARD, RICHARD J.. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications. INTERNATIONAL JOURNAL OF GENOMICS, v. 2018, p. 12-pg., 2018-01-01. (16/18827-7, 16/20358-5)

RIBEIRO, LUCAS F.; RIBEIRO, LILIANE F. C.; BARRETO, MATHEUS Q.; WARD, RICHARD J.. Protein Engineering Strategies to Expand CRISPR-Cas9 Applications. INTERNATIONAL JOURNAL OF GENOMICS, 2018. (16/18827-7, 16/20358-5)

RIBEIRO, LUCAS FERREIRA; LOPES, ERICA M.; KISHI, LUCIANO T.; COSTA RIBEIRO, LILIANE FRAGA; MENEGUETI, MAYRA GONCALVES; GASPAR, GILBERTO GAMBERO; SILVA-ROCHA, RAFAEL; GUAZZARONI, MARIA-EUGENIA. Microbial Community Profiling in Intensive Care Units Expose Limitations in Current Sanitary Standards. FRONTIERS IN PUBLIC HEALTH, v. 7, AUG 28 2019. (15/04309-1, 16/20358-5, 16/18827-7)

RIBEIRO, LUCAS F.; AMARELLE, VANESA; RIBEIRO, LILIANE F. C.; GUAZZARONI, MARIA-EUGENIA. Converting a Periplasmic Binding Protein into a Synthetic Biosensing Switch through Domain Insertion. BIOMED RESEARCH INTERNATIONAL, v. 2019, p. 15-pg., 2019-01-01. (16/18827-7, 16/20358-5, 15/04309-1)

RIBEIRO, LILIANE F. C.; CHELIUS, CYNTHIA; BOPPIDI, KARTHIK R.; NAIK, NISHA S.; HOSSAIN, SIMIN; RAMSEY, JESSICA J. J.; KUMAR, JYOTHI; RIBEIRO, LUCAS F.; OSTERMEIER, MARC; BAO TRAN; et al. Comprehensive Analysis of Aspergillus nidulans PKA Phosphorylome Identifies a Novel Mode of CreA Regulation. MBIO, v. 10, n. 2, p. 13-pg., 2019-03-01. (16/18827-7, 16/20358-5)