A bio-inspired cell-free system for cannabinoid production from inexpensive inputs
- 24 August 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Chemical Biology
- Vol. 16 (12), 1427-1433
- https://doi.org/10.1038/s41589-020-0631-9
Abstract
Moving cannabinoid production away from the vagaries of plant extraction and into engineered microbes could provide a consistent, purer, cheaper and environmentally benign source of these important therapeutic molecules, but microbial production faces notable challenges. An alternative to microbes and plants is to remove the complexity of cellular systems by employing enzymatic biosynthesis. Here we design and implement a new cell-free system for cannabinoid production with the following features: (1) only low-cost inputs are needed; (2) only 12 enzymes are employed; (3) the system does not require oxygen and (4) we use a nonnatural enzyme system to reduce ATP requirements that is generally applicable to malonyl-CoA-dependent pathways such as polyketide biosynthesis. The system produces ~0.5 g l−1 cannabigerolic acid (CBGA) or cannabigerovarinic acid (CBGVA) from low-cost inputs, nearly two orders of magnitude higher than yeast-based production. Cell-free systems such as this may provide a new route to reliable cannabinoid production.Keywords
Funding Information
- DOE | Advanced Research Projects Agency - Energy (DE-AR0000556)
- DOE | SC | Biological and Environmental Research (DE-FC02-02ER63421)
This publication has 36 references indexed in Scilit:
- The cannabinoid Δ9-tetrahydrocannabivarin (THCV) ameliorates insulin sensitivity in two mouse models of obesityNutrition & Diabetes, 2013
- Identification of olivetolic acid cyclase from Cannabis sativa reveals a unique catalytic route to plant polyketidesProceedings of the National Academy of Sciences of the United States of America, 2012
- Cell-free synthetic biology: Thinking outside the cellMetabolic Engineering, 2012
- eQuilibrator--the biochemical thermodynamics calculatorNucleic Acids Research, 2011
- Five hard truths for synthetic biologyNature, 2010
- Characterization of olivetol synthase, a polyketide synthase putatively involved in cannabinoid biosynthetic pathwayFEBS Letters, 2009
- Bioreactor scale-up and oxygen transfer rate in microbial processes: An overviewBiotechnology Advances, 2009
- Functions of Malonate Decarboxylase Subunits fromPseudomonas putidaBioscience, Biotechnology, and Biochemistry, 2003
- A convenient synthesis of disodium acetyl phosphate for use in in situ ATP cofactor regenerationThe Journal of Organic Chemistry, 1983
- Cannabis. X. The isolation and structures of four new propyl cannabinoid acids, tetrahydrocannabivarinic acid, cannabidivarinic acid, cannabichromevarinic acid and cannabigerovarinic acid, from Thai Cannabis, 'Meao variant'.CHEMICAL & PHARMACEUTICAL BULLETIN, 1977