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Construction of a microbial biofactory for production of 3-hydroxypropionic acid using CRISPR/Cas system

Grant number: 21/12060-4
Support Opportunities:Scholarships in Brazil - Scientific Initiation
Effective date (Start): February 01, 2022
Effective date (End): February 28, 2023
Field of knowledge:Biological Sciences - Genetics - Molecular Genetics and Genetics of Microorganisms
Principal Investigator:Nádia Maria Vieira Sampaio
Grantee:Leonardo Blankenburg Lins Barroso Arruda
Host Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia e Inovações (Brasil). Campinas , SP, Brazil

Abstract

The global economy is heavily dependent on the petrochemical industry, which has contributed to worsening climate change. In this context, the development of microorganisms capable of producing renewable biochemicals is essential for the development of a sustainable bioeconomy. Aiming to contribute to the solution of this challenge, this project aims to develop a strain of Saccharomyces cerevisiae capable of producing 3-hydroxypropionic acid (3-HP) from glucose, a renewable and abundant substrate. 3-HP acid is considered a high value-added platform chemical, as it can be converted into several derivatives with applications in the production of paints, superabsorbent materials, plastics, among others. For this purpose, a collection of six laboratory and industrial strains will be evaluated in 3-HP tolerance tests to determine the most promising one to be used as a production platform. The strain design, already in progress, combines literature data, as well as data from in silico optimization using genomic-scale metabolic models to identify genetic modifications, not yet described, with the potential to enhance 3-HP biosynthesis via malonyl-CoA pathway. The construction of the producing strain will be carried out in stages using the CRISPR-Cas technique to integrate the genes of interest into the yeast genome. Finally, the impact of these genetic modifications on the growth and stability of the final strain will be evaluated by monitoring variations in growth and production rates over 80 generations. It is expected that the results obtained will contribute to the development of a proof-of-concept strain to produce 3-HP that will serve as a basis for future optimization projects. (AU)

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