Development of a cannabinoid yeast factory: improving geranyl pyrophosphate production through metabolic engineering

Abstract As the population ages there is a growing concern about the impact of neurological diseases on the healthcare system Neurological diseases e g Alzheimer s Disease are the fastest growing cause of death in the United States and the development of new drugs has been very slow Natural products are powerful sources for the discovery of new medications recently cannabinoids have been identified as a general class of natural compounds with a wide range of proven therapeutic effects including treatment of neurodegenerative diseases In fact studies of cannabinoids led to the discovery of the endocannabinoid system ECS regarded as one of the most important modulators of nervous system processes and a targetable system for neurological diseases Since then cannabinoids have been found to have efficacy in treating pain neuroinflammation and bone loss among many other conditions Cannabinoids can be found in many plants the most well known is Cannabis sativa from which a variety of cannabinoids have been isolated and shown to be medically relevant However it is difficult to source cannabinoids in high purity inhibiting research and making large scale commercial production problematic A biosynthetic approach to producing these natural products not only addresses these problems but allows a detailed understanding of the natural product chemistry furthering medical research and pharmaceutical development into the ECS To enable the development of natural cannabinoids as therapeutic agents Librede has developed a biosynthetic cannabinoid production drug discovery platform by genetically engineering yeast with selected portions of the Cannabis sativa metabolic pathways The production of cannabinoids in yeast is an ideal platform because fermentation and genetic engineering are well established low cost and scalable Librede s biosynthetic cannabinoid production platform is modular by adding or removing expression of different enzymes different cannabinoids can be produced as desired even cannabinoids produced in low abundance naturally Using this approach in preliminary work we have produced Cannabidiolic acid CBDA in yeast the world s first biosynthetically produced cannabinoids outside of plants Although this proof of concept work showed that cannabinoids can be produced by yeast the yield of CBDA production was limited by the availability of a specific chemical intermediate geranyl pyrophosphate GPP In order to make our platform feasible for research and cannabinoid production we will improve GPP levels by reducing its diversion to alternative products by knocking out or down regulate diverting metabolic pathways as well as increasing GPP production Narrative Cannabinoids have been shown to be efficacious to treat multiple sclerosis and other neurodegenerative diseases but they are produced in plants in small amounts Librede has developed the world s first in vivo biosynthetic platform for cannabinoid production from genetically engineered yeast We propose to increase the yield of cannabinoid production in our platform by engineering yeast to increase the production of geranyl pyrophosphate GPP a biochemical intermediate in cannabinoid biosynthesis