Characterization of resistance to high osmolarity in Pseudomonas putida using synthetic circuits

• Grant number: 25/13896-0
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• Start date: September 01, 2025
• End date: August 31, 2026
• Field of knowledge: Biological Sciences - Microbiology - Biology and Physiology of Microorganisms
• Principal Investigator: María Eugenia Guazzaroni
• Grantee: Jhenyfer Teodoro Genesio
• Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated research grant: 21/01748-5 - Discovery and development of new non-model bacterial chassis for biotechnological applications, AP.TEM

Abstract

Microorganisms that inhabit extreme environments have significant genetic potential, adapting to variations in temperature, radiation, salinity, acidity and pressure. The study of microbial diversity, focusing on the metabolic activities of these organisms, allows us to identify biochemical strategies essential for survival, driving advances in industrial processes. In this context, the use of microorganisms in bioprocesses is highly advantageous, as it enables the conversion of low-value substances into valuable products. However, the high demand for fresh water in these operations results in a water footprint that is harmful to the environment. A promising alternative is the use of seawater, which represents the majority of global water resources. However, the high concentration of salts in this environment poses challenges to the growth and viability of microorganisms, making it essential to identify strains resistant to this condition.This study aims to investigate whether genes previously identified in metagenomes from extreme environments confer resistance to high osmolarity in bacteria. To this end, we propose to evaluate the functionality of these genes in synthetic circuits using Pseudomonas putida, a bacterium of industrial relevance. Genes previously characterized by our research group in Escherichia coli, using a high-copy-number vector, will be cloned and transferred to P. putida through the pVANT plasmid. We will then conduct growth experiments and salinity tolerance assays in the presence of NaCl, analysing kinetic parameters to identify genetic circuits that confer efficient resistance to saline stress in this bacterium.