CONICET PhD fellow
Director: José Luis Barra
PhD Thesis Topic
Metabolic engineering in lactic acid bacteria
The main objective of this project is to develop genetic and biotechnological tools to accomplish the use of Lactic Acid Bacteria (LAB), known as GRAS (Generally Recognized As Safe) organisms, as cell factories for the production of high-value metabolites for food and Biotech industries, either by cloning single genes or metabolic pathways into plasmids or into the genome. As specific objectives we propose:
1. Construction of a shuttle promoter library for Escherichia coli and Lactococcus lactis using L. lactis genomic fragments with promoter activity, which could lead different levels of expression of genes or metabolic pathways of interest.
2. Design and implementation of polygenic expression systems of metabolic pathways in LAB. Aiming the production of secondary metabolites of interest in LAB, we will study the use of suitable promoter regions, RBS and codon usage, among others, into plasmidic constructions. We will express carotenoids as a case study.
3. Design of CRISPR-Cas9 system for efficient LAB genome editing: We will put together into a single plasmid all the elements to implement CRISPR-Cas9 system for L. lactis genome editing. We expect not only to develop new key tools to use LAB as cell factories but also to accomplish the genetic improvement of industrial LAB strains.