Engineering a microbial platform for de novo biosynthesis of diverse methylxanthines |
| |
| Affiliation: | 1. Department of Bioengineering, Stanford University, 443 Via Ortega, MC 4245, Stanford, CA 94305, United States;2. Synthetic Genomics, La Jolla, CA 92037, United States;1. Division of Medical Oncology, Department of Internal Medicine, The Ohio State University College of Medicine, 410 W 12th Avenue, Columbus, OH, 43210, USA;2. The Ohio State University Comprehensive Cancer Center - Arthur G. James Cancer Hospital and Richard J. Solove Institute, 410 W 12th Avenue, Columbus, OH, 43210, USA;3. Department of Nutrition, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA, 02115, USA;4. Division of Women''s Health, Brigham and Women''s Hospital, 75 Francis Street, Boston, MA, 02115, USA;5. Department of Epidemiology, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA, 02115, USA;6. Department of Biostatistics, Harvard T.H. Chan School of Public Health, 677 Huntington Avenue, Boston, MA, 02115, USA;7. Channing Division of Network Medicine, Brigham and Women''s Hospital, 181 Longwood Avenue, Boston, MA, 02115, USA;8. Department of Biostatistics and Epidemiology, University of Massachusetts-Amherst, 715 N Pleasant Street, Amherst, MA, 01003, USA;9. Department of Medicine, Harvard Medical School, 25 Shattuck Street, Boston, MA, 02115, USA;10. Division of Preventive Medicine, Brigham and Women''s Hospital, 900 Commonwealth Avenue, Boston, MA, 02115, USA;11. Division of Research, Kaiser Permanente Northern California, 2051 Franklin Street, Oakland, CA, 94612, USA;12. Department of Social Sciences and Health Policy, Wake Forest School of Medicine, 475 Vine Street, Winston-Salem, NC, 27101, USA;13. Department of Preventive Medicine, Northwestern University, 680 N Lakeshore Dr, Chicago, IL, 60611, USA;14. Broad Institute of the Massachusetts Institute of Technology and Harvard University, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA;1. Key laboratory of Biomass Chemical Engineering (Ministry of Education), College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;2. Xiangya International Academy of Translational Medicine, Central South University, Changsha 410013, Hunan, China;1. Department of Chemical and Biomolecular Engineering, National University of Singapore, 4 Engineering Drive 4, Singapore 117585, Singapore;2. Temasek Laboratories, National University of Singapore, T-Lab Building 5A, Engineering Drive 1, Singapore 117411, Singapore;3. Singapore Institute of Technology, 10 Dover Drive, Singapore 138683, Singapore;1. College of Chemistry and Chemical Engineering, Chemical Synthesis and Pollution Control Key Laboratory of Sichuan Province, China West Normal University, Nanchong 637002, People’s Republic of China;2. College of Chemistry, Key Laboratory of Green Chemistry and Technology in Ministry of Education, Sichuan University, Chengdu 610064, People’s Republic of China;1. Rudjer Boskovic Institute, Division for Marine and Environmental Research, Bijenicka c. 54, 10000 Zagreb, Croatia;2. IRCCS – Istituto di Ricerche Farmacologiche “Mario Negri”, Department of Environmental Health Sciences, Via La Masa 19, 20156 Milan, Italy |
| |
| Abstract: | Engineered microbial biosynthesis of plant natural products can support manufacturing of complex bioactive molecules and enable discovery of non-naturally occurring derivatives. Purine alkaloids, including caffeine (coffee), theophylline (antiasthma drug), theobromine (chocolate), and other methylxanthines, play a significant role in pharmacology and food chemistry. Here, we engineered the eukaryotic microbial host Saccharomyces cerevisiae for the de novo biosynthesis of methylxanthines. We constructed a xanthine-to-xanthosine conversion pathway in native yeast central metabolism to increase endogenous purine flux for the production of 7-methylxanthine, a key intermediate in caffeine biosynthesis. Yeast strains were further engineered to produce caffeine through expression of several enzymes from the coffee plant. By expressing combinations of different N-methyltransferases, we were able to demonstrate re-direction of flux to an alternate pathway and develop strains that support the production of diverse methylxanthines. We achieved production of 270 μg/L, 61 μg/L, and 3700 μg/L of caffeine, theophylline, and 3-methylxanthine, respectively, in 0.3-L bench-scale batch fermentations. The constructed strains provide an early platform for de novo production of methylxanthines and with further development will advance the discovery and synthesis of xanthine derivatives. |
| |
| Keywords: | Plant natural products Purine alkaloids Methylxanthines Synthetic biology Yeast |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
