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Complete biosynthesis of cannabinoids and their unnatural analogues in yeast

Journal Article · · Nature (London)
Cannabis sativa L. has been cultivated and used around the globe for its medicinal properties for millennia. Some cannabinoids, the hallmark constituents of Cannabis, and their analogues have been investigated extensively for their potential medical applications. Certain cannabinoid formulations have been approved as prescription drugs in several countries for the treatment of a range of human ailments. However, the study and medicinal use of cannabinoids has been hampered by the legal scheduling of Cannabis, the low in planta abundances of nearly all of the dozens of known cannabinoids, and their structural complexity, which limits bulk chemical synthesis. Here we report the complete biosynthesis of the major cannabinoids cannabigerolic acid, Δ9-tetrahydrocannabinolic acid, cannabidiolic acid, Δ9-tetrahydrocannabivarinic acid and cannabidivarinic acid in Saccharomyces cerevisiae, from the simple sugar galactose. To accomplish this, we engineered the native mevalonate pathway to provide a high flux of geranyl pyrophosphate and introduced a heterologous, multi-organism-derived hexanoyl-CoA biosynthetic pathway. We also introduced the Cannabis genes that encode the enzymes involved in the biosynthesis of olivetolic acid, as well as the gene for a previously undiscovered enzyme with geranylpyrophosphate:olivetolate geranyltransferase activity and the genes for corresponding cannabinoid synthases. Furthermore, we established a biosynthetic approach that harnessed the promiscuity of several pathway genes to produce cannabinoid analogues. Feeding different fatty acids to our engineered strains yielded cannabinoid analogues with modifications in the part of the molecule that is known to alter receptor binding affinity and potency. We also demonstrated that our biological system could be complemented by simple synthetic chemistry to further expand the accessible chemical space. Our work presents a platform for the production of natural and unnatural cannabinoids that will allow for more rigorous study of these compounds and could be used in the development of treatments for a variety of human health problems.
Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); Sandia National Laboratories (SNL-CA), Livermore, CA (United States)
Sponsoring Organization:
Jiangnan University Graduate Scholarship; National Science Foundation (NSF); USDOE Office of Science (SC)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1766485
Alternate ID(s):
OSTI ID: 1868480
Journal Information:
Nature (London), Journal Name: Nature (London) Journal Issue: 7746 Vol. 567; ISSN 0028-0836
Publisher:
Nature Publishing GroupCopyright Statement
Country of Publication:
United States
Language:
English

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Additional file 15 of Integrated multi-omics analysis and microbial recombinant protein system reveal hydroxylation and glycosylation involving nevadensin biosynthesis in Lysionotus pauciflorus dataset January 2022
Additional file 1 of Integrated multi-omics analysis and microbial recombinant protein system reveal hydroxylation and glycosylation involving nevadensin biosynthesis in Lysionotus pauciflorus dataset January 2022
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Additional file 3 of Integrated multi-omics analysis and microbial recombinant protein system reveal hydroxylation and glycosylation involving nevadensin biosynthesis in Lysionotus pauciflorus dataset January 2022
Additional file 5 of Integrated multi-omics analysis and microbial recombinant protein system reveal hydroxylation and glycosylation involving nevadensin biosynthesis in Lysionotus pauciflorus dataset January 2022
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Baker's Yeast Catalyzed Multicomponent Reactions: A New Hope? journal November 2019
The renaissance of yeasts as microbial factories in the modern age of biomanufacturing journal September 2019
Production of semi-biosynthetic nepetalactone in yeast journal October 2019
Complex Patterns of Cannabinoid Alkyl Side-Chain Inheritance in Cannabis journal August 2019
Computer-aided re-engineering of nonribosomal peptide and polyketide biosynthetic assembly lines journal January 2019
Genetic platforms for heterologous expression of microbial natural products journal January 2019
Applied evolution: phylogeny-based approaches in natural products research journal January 2019
Impacts of cannabinoid epigenetics on human development: reflections on Murphy et. al. ‘cannabinoid exposure and altered DNA methylation in rat and human sperm’ epigenetics 2018; 13: 1208-1221. journal June 2019
Enhanced scale and scope of genome engineering and regulation using CRISPR/Cas in Saccharomyces cerevisiae journal October 2019
Design of a programmable biosensor-CRISPRi genetic circuits for dynamic and autonomous dual-control of metabolic flux in Bacillus subtilis journal December 2019
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Superloser: A Plasmid Shuffling Vector for Saccharomyces cerevisiae with Exceedingly Low Background journal August 2019
Impact of Nitrogen Nutrition on Cannabis sativa: An Update on the Current Knowledge and Future Prospects journal November 2019

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