Turning the Screw: Engineering Extreme pH Resistance in Escherichia coli through Combinatorial Synthetic Operons

Author(s)

de Siqueira, V, Guilherme M. ; Silva-Rocha, Rafael ^[1] ; Guazzaroni, Maria-Eugenia ^[2]

Affiliation

^[1] Fac Med Ribeirao Preto FMRP USP, Dept Biol Celular & Mol & Bioagentes Patog, BR-14049900 Ribeirao Preto, SP - Brazil

^[2] Fac Filosofia Crencias & Letras Ribeirao Preto FF, Dept Biol, BR-14040901 Ribeirao Preto, SP - Brazil

• Document type: Journal article
• Source: ACS SYNTHETIC BIOLOGY; v. 9, n. 6, p. 1254-1262, JUN 19 2020.
• Web of Science Citations: 0

Abstract

Adoption of microorganisms as platforms for sustainable biobased production requires host cells to be able to withstand harsh conditions, usually very distant from those in which these organisms are naturally adapted to thrive. However, novel survival mechanisms unearthed by the study of microbiomes from extreme habitats may be exploited to enhance microbial robustness under the strict conditions needed for different industrial appplications. In this work, synthetic biology approaches were used to engineer enhanced acidic resistance in Escherichia coli through the characterization of a collection of unique operons composed of combinatorial assemblies of three novel genes from an extreme environment and three synthetic ribosome binding sites. The results here presented illustrate the efficacy of combining different metagenomic genes for resistance in synthetic operons, as expression of these gene clusters increased hundred-fold the survival percentage of cells exposed to an acidic shock in minimal media at pH 1.9 under aerobic conditions. (AU)

• FAPESP's process: 15/04309-1 - Novel approaches to improve functional screening of biocatalysts in metagenomic libraries
• Grantee: María Eugenia Guazzaroni
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 18/07261-8 - Expanding acid resistance in bacteria using synthetic circuits
• Grantee: Guilherme Marcelino Viana de Siqueira
• Support Opportunities: Scholarships in Brazil - Master