Heterologous large-scale production of populninic acid in Nicotiana benthamiana system via combinatorial biosynthesis using P450 enzymes and the substrate maytenoic acid

• Grant number: 23/04093-5
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: September 01, 2023
• End date: August 31, 2024
• Field of knowledge: Physical Sciences and Mathematics - Chemistry - Organic Chemistry
• Principal Investigator: Maysa Furlan
• Grantee: Otávio Aguiar de Souza
• Supervisor: Vincent Courdavault
• Host Institution: Instituto de Química (IQ). Universidade Estadual Paulista (UNESP). Campus de Araraquara. Araraquara , SP, Brazil
• Institution abroad: Université François-Rabelais, France
• Associated to the scholarship: 21/05646-2 - Metabolic engineering applied to the elucidation of the biosynthetic pathway and sustainable production of quinonemethide triterpenes via heterologous expression in Saccharomyces cerevisiae, BP.DR

Abstract

Recently we identified in the heterologous system Saccharomyces cerevisiae that three cytochrome P450 enzymes catalyze the oxidation of maytenoic acid (a biosynthetic precursor of quinonemethide triterpenes or QMTs in the plant species Maytenus ilicifolia, as our group proved in 2019) to the putative populninic acid (FAPESP Grant 21/05646-2). This oxidation step may be vital for further elucidation of the complete metabolic pathway of QMTs, but to unequivocally prove the role of the P450 enzymes acting in this oxidation step, it is appropriate to obtain the oxidized derivative in appreciable quantities, which is not always possible using yeast platforms because that would require scaling up the process, and this task is not always trivial. Thus, this project aims at the large-scale production of populninic acid via combinatorial biosynthesis using as the alternative heterologous host the plant species Nicotiana benthamiana. The project was designed to be developed during 6 months in the research group of Professor Vincent Courdavault (Université de Tours, BBV - EA2106 Biomolécules et Biotechnologies Végétales, Tours, France), which has a consolidated experience in combinatorial biosynthesis, metabolic engineering, synthetic biology of plant specialized metabolites and use of N. benthamiana system for the straightforward and optimized large-scale production of such compounds. Additionally, Professor Vincent Courdavault has a laboratory that offers all facilities including a modern analytical platform that will enable the unequivocal identification of the compounds produced and, therefore, the potential role of P450 enzymes in this oxidation step. This will also enable the renewable and optimized production of a derivative of maytenoic acid with interesting pharmacological properties. (AU)