Engineering microbes for the production of valuable natural products is often hindered by the regulation of native competing metabolic networks in host. This is particularly evident in the case of terpenoid synthesis in yeast, where the canonical terpenoid precursors are tightly coupled to the biosynthesis of sterols essential for yeast viability. One way to circumvent this limitation is by engineering product pathways less connected to the host native metabolism. Here, we introduce a two-step isopentenol utilization pathway (IUP) in Saccharomyces cerevisiae to augment the native mevalonate pathway by providing a shortcut to the synthesis of the common terpenoid precursors, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). As such, the IUP was capable of elevating the IPP/DMAPP pool by 147-fold compared with the native pathway. We further demonstrate that cofeeding isoprenol and prenol enhances geranyl diphosphate (GPP) content for monoterpene biosynthesis. More importantly, we established a synthetic three-step route for efficient synthesis of di-and tetraterpene precursor geranylgeranyl diphosphate (GGPP), circumventing the competition with farnesyl diphosphate (FPP) for sterol biosynthesis and elevating the GGPP level by 374-fold. We combine these IUP-supported precursor-forming platforms with downstream terpene synthases to harness their potential and improve the production of industrially relevant terpenoids by several fold. In conclusion, our exploration provides a universal and effective platform for supporting terpenoid synthesis in yeast.
Ma, Yongshuo, et al. "Engineering a universal and efficient platform for terpenoid synthesis in yeast." Proceedings of the National Academy of Sciences of the United States of America, vol. 120, no. 1, Dec. 2022. https://doi.org/10.1073/pnas.2207680120
Ma, Yongshuo, Zu, Yuexuan, Huang, Sanwen, & Stephanopoulos, Gregory (2022). Engineering a universal and efficient platform for terpenoid synthesis in yeast. Proceedings of the National Academy of Sciences of the United States of America, 120(1). https://doi.org/10.1073/pnas.2207680120
Ma, Yongshuo, Zu, Yuexuan, Huang, Sanwen, et al., "Engineering a universal and efficient platform for terpenoid synthesis in yeast," Proceedings of the National Academy of Sciences of the United States of America 120, no. 1 (2022), https://doi.org/10.1073/pnas.2207680120
@article{osti_2417703,
author = {Ma, Yongshuo and Zu, Yuexuan and Huang, Sanwen and Stephanopoulos, Gregory},
title = {Engineering a universal and efficient platform for terpenoid synthesis in yeast},
annote = {Engineering microbes for the production of valuable natural products is often hindered by the regulation of native competing metabolic networks in host. This is particularly evident in the case of terpenoid synthesis in yeast, where the canonical terpenoid precursors are tightly coupled to the biosynthesis of sterols essential for yeast viability. One way to circumvent this limitation is by engineering product pathways less connected to the host native metabolism. Here, we introduce a two-step isopentenol utilization pathway (IUP) in Saccharomyces cerevisiae to augment the native mevalonate pathway by providing a shortcut to the synthesis of the common terpenoid precursors, isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). As such, the IUP was capable of elevating the IPP/DMAPP pool by 147-fold compared with the native pathway. We further demonstrate that cofeeding isoprenol and prenol enhances geranyl diphosphate (GPP) content for monoterpene biosynthesis. More importantly, we established a synthetic three-step route for efficient synthesis of di-and tetraterpene precursor geranylgeranyl diphosphate (GGPP), circumventing the competition with farnesyl diphosphate (FPP) for sterol biosynthesis and elevating the GGPP level by 374-fold. We combine these IUP-supported precursor-forming platforms with downstream terpene synthases to harness their potential and improve the production of industrially relevant terpenoids by several fold. In conclusion, our exploration provides a universal and effective platform for supporting terpenoid synthesis in yeast.},
doi = {10.1073/pnas.2207680120},
url = {https://www.osti.gov/biblio/2417703},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {ISSN 0027-8424},
number = {1},
volume = {120},
place = {United States},
publisher = {National Academy of Sciences},
year = {2022},
month = {12}}
Massachusetts Institute of Technology (MIT), Cambridge, MA (United States)
Sponsoring Organization:
USDOE Office of Science (SC); National Key Research and Development Program of China
Grant/Contract Number:
SC0022016
OSTI ID:
2417703
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 1 Vol. 120; ISSN 0027-8424