Investigation of malonyl-CoA reductase enzyme variants for optimizing bioproduction of 3-hydroxypropionic acid in yeast

• Grant number: 23/00885-4
• Support Opportunities: Scholarships in Brazil - Master
• Start date: February 01, 2024
• End date: February 28, 2025
• 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 Inovação (Brasil). Campinas , SP, Brazil
• Associated research grant: 20/16759-0 - Synthetic biology tools for scalable production of biorenewable 3-hydroxypropionic acid, AP.JP

Abstract

The development of new biotechnological production routes is crucial for the development of a sustainable bioeconomy that consumes fewer petrochemical resources and combats the worsening effects of climate change. 3-Hydroxypropionic acid (3-HP) is considered a platform chemical, capable of being chemically converted into a series of molecules with commercial applications, such as acrylates, 1,3-propanediol, among others. Therefore, this research project seeks to build a strain of Saccharomyces cerevisiae that produces 3-HP from glucose, a renewable and abundant substrate. Studies aimed at the biological production of 3-HP from the malonyl-CoA precursor have mainly used the bifunctional malonyl-CoA reductase (MCR) enzyme from Chloroflexus aurantiacus and reported limited enzymatic activity. Therefore, this project aims to investigate potentially more efficient MCR variants of Chloroflexus aurantiacus (CaMCR). Orthologs will be mapped through a Sequence Similarity Network (SSN) and five of them will be analyzed in in vivo assays in Saccharomyces cerevisiae and in vitro. Furthermore, the C- and N-terminal portions of the C. aurantiacus bifunctional MCR will be uncoupled and expressed at different levels in order to determine an optimized balance of each enzymatic step required for 3-HP production. The results obtained are expected to contribute to the development of an MCR enzyme with greater enzymatic activity for the production of 3-HP, which will serve as a basis for future optimization projects in other aspects of the yeast strain.