Evolutionary engineering of Escherichia coli for anaerobic production of polyhydroxyalkanoates from sugarcane-derived sucrose

• Grant number: 20/02246-0
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• Start date: June 01, 2020
• End date: May 31, 2024
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Agreement: Netherlands Organisation for Scientific Research (NWO)
• Principal Investigator: Jonas Contiero
• Grantee: Joneclei Alves Barreto
• Host Institution: Instituto de Pesquisa em Bioenergia (IPBEN). Universidade Estadual Paulista (UNESP). Campus de Rio Claro. Rio Claro , SP, Brazil
• Associated research grant: 17/50249-6 - A novel strategy for anaerobic production of polyhydroxyalkanoates (PHA), AP.TEM

Abstract

Adaptive Laboratory Evolution (ALE) is a powerful tool to shape microorganisms for industrial applications. This project aims at using ALE for optimizing Escherichia coli to growth under anaerobic conditions and to use sucrose as a carbon source. For these purposes, E. coli strains will be first engineered with the csc genes (cscB, cscK, cscA), for uptake of sucrose, and with the gene sucP, for sucrose phosphorolysis. Genetically modified strains will be then challenged (ALE approach) to propagate under anaerobic conditions and to consume increasing amounts of sucrose from synthetic medium. Mutations that accumulated during ALE will be discovered by next-generation sequencing of the genomes and then reverse engineered into the parental strains. The resulting strains will be further submitted to ALE for propagation in sugarcane-derived molasses and juices. Finally, strains that were optimized for anaerobic growth on sucrose will be subjected to Metabolic Engineering for polyhydroxyalkanoates (PHAs) synthesis, resulting in ideal strains for bioplastic production from sugarcane feedstock. (AU)