A comprehensive platform for sustainable biosynthesis of rare natural products in Pichia pastoris
Abstract
Plant natural products (NPs) have significantly contributed to the drug discovery field, especially for cancer and infectious diseases. However, their low abundance in plants has limited their extraction, while their chemical synthesis is challenging due to the complex structure of NPs. The Food and Agricultural Organization of the United Nations has estimated that global food production must increase by 70%–100% by 2050, when the world population is predicted to reach 9 billion (2 billion additional people on the planet within 30 years) (FAO, 2009). As another consequence of population growth, the population of older people will increase. By 2050, 130 million people will have osteoarthritis (Mobasheri et al., 2019) demanding pain-relieving NP drugs, and 20 million new cancer cases will occur requiring NP-based anticancer agents for treatment.
To obtain diverse bioactive NP-derived compounds with high yields and to sufficiently produce rare NPs, the feedback regulatory network controlling the expression of the biosynthetic pathway genes must be well understood. COMPLATn - COMprehensive PLATform for sustainable biosynthesis of NP - aims to enable yeast P. pastoris to synthesize the optimized level of plant NPs as an alternative to nature. To this end, we will establish artificial regulators to control the time and expression level of biosynthetic pathway enzymes in the presence of light in yeast. We will also establish a combinatorial optimization strategy to discover the optimal expression level for the individual enzymes. The yeast cells producing the maximized amount of NP will be screened using biological sensors. The platform is united with complementary automatic bioprocessing (to stabilize the productivity of NPs) and droplet-based single-cell analysis techniques (to identify the genetic identity of NP producers). As a result, COMPLATn generates a huge dataset linking the genetic identity and productivity of the system to support the understanding of feedback regulation of NP biosynthetic pathways and the discovery of their key building blocks.
As a proof of concept, we will employ COMPLATn for sustainable biomanufacturing of strictosidine, positioning yeast as a scalable system to produce more than 3,000 natural NPs, and we will convert it to the anticancer small molecule drugs catharanthine, vindoline, and vinblastine directly in yeast, allowing us to find out the key building blocks in the long biosynthetic pathways of alkaloids, an important group of NPs.
Principal investigators
Participating organisational units of HU Berlin
Financer
DFG Emmy Noether Programm
Duration of project
Start date: 04/2024
End date: 03/2030
