Awakening of cryptic biosynthetic gene clusters using obese red yeast

Research Project, 2019 – 2023

Natural products are indispensable in modern medicine, representing over 32% among all new approved drugs. However, most organisms harbouring the required biosynthetic gene clusters (BGCs) are unculturable in the lab, or their BGCs are transcriptionally silent, precluding biosynthesis and further analysis of these unique compounds. In this project we will awaken BGCs by their expression in a heterologous host, with a focus on the classes of natural products known as polyketides and meroterpenoids.The red obese yeast Rhodosporidium toruloides is a potent host for BGC production, as it is a prolific producer of malonyl-CoA and farnesyl-PP, two key precursors required for polyketide and meroterpenoid biosynthesis. To enhance heterologous production of complex natural products, we will first further enhance malonyl-CoA and farnesyl-PP supply in R. toruloides through metabolic engineering. We have developed a computational model that can determine the optimal protein allocation for a certain cellular objective, considering kinetics, protein size and quantitative proteomics measurements. Guided by these model predictions we will then genetically modify the expression of key proteins.Subsequently we will use the reconstructed chassis strain for polyketide and meroterpenoid biosynthesis for heterologous production. Initially using BGCs with known function to validate our methods, we will then embark on unravelling cryptic BGCs, to discover future´s drugs.