Metabolic Engineering of the acetate-evolved Synechocystis sp. B12 for improved PHB production

• Grant number: 24/05662-6
• Support Opportunities: Scholarships abroad - Research Internship - Post-doctor
• Start date: December 18, 2024
• End date: December 17, 2025
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Engineering
• Principal Investigator: Cassius Vinicius Stevani
• Grantee: Dielle Pierotti Procópio
• Supervisor: Tomas Morosinotto
• Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Institution abroad: Università degli Studi di Padova, Italy
• Associated to the scholarship: 23/07986-0 - Adaptive laboratory evolution of the PHB-producing Synechocystis sp. PCC6803 for improved acetate consumption, BP.PD

Abstract

The cyanobacterium Synechocystis sp. B12, initially isolated from the mangrove habitat of Brazil, has emerged as a proficient producer of a specialized derivative of polyhydroxyalkanoates (PHA), specifically poly(3-hydroxybutyrate) (PHB), exhibiting an impressive yield of 31% cell dry weight (cdw) under light intensity of 300 ¼mol photons/m2s1. This distinctive characteristic presents substantial promise for biotechnological exploitation, particularly in directly synthesizing PHB from CO2. In pursuit of enhancing PHB accumulation in the B12 strain, this investigation aims to explore the genetic manipulation and metabolic engineering of an acetate-evolved Synechocystis sp. B12 for augmented PHB production. Rational metabolic engineering approaches will be deployed to integrate the PHB biosynthetic pathway genes (phaA, phaB, phaE, and phaC) into the neutral locus of acetate-adapted Synechocystis sp. B12, derived from the ongoing postdoctoral endeavor of the current researcher (FAPESP 202307986-0). To facilitate the heightened expression of the PHB biosynthetic pathway in Synechocystis sp. B12, plasmids PEERM3_phaABEC will be engineered by amplifying the gene sequences with overhangs homologous to the PEERM3 plasmid.