Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast
Abstract Background Metabolic rewiring in microbes is an economical and sustainable strategy for synthesizing valuable natural terpenes. Terpenes are the largest class of nature-derived specialized metabolites, and many have valuable pharmaceutical or biological activity. Squalene, a medicinal terpe...
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| Format: | Article |
| Language: | English |
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BMC
2022-10-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-022-02208-9 |
| _version_ | 1798029965919256576 |
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| author | So-Hee Son Jae-Eung Kim Soo Young Moon In-Seung Jang Byung Jo Yu Ju Young Lee |
| author_facet | So-Hee Son Jae-Eung Kim Soo Young Moon In-Seung Jang Byung Jo Yu Ju Young Lee |
| author_sort | So-Hee Son |
| collection | DOAJ |
| description | Abstract Background Metabolic rewiring in microbes is an economical and sustainable strategy for synthesizing valuable natural terpenes. Terpenes are the largest class of nature-derived specialized metabolites, and many have valuable pharmaceutical or biological activity. Squalene, a medicinal terpene, is used as a vaccine adjuvant to improve the efficacy of vaccines, including pandemic coronavirus disease 2019 (COVID-19) vaccines, and plays diverse biological roles as an antioxidant and anticancer agent. However, metabolic rewiring interferes with inherent metabolic pathways, often in a way that impairs the cellular growth and fitness of the microbial host. In particular, as the key starting molecule for producing various compounds including squalene, acetyl-CoA is involved in numerous biological processes with tight regulation to maintain metabolic homeostasis, which limits redirection of metabolic fluxes toward desired products. Results In this study, focusing on the recycling of surplus metabolic energy stored in lipid droplets, we show that the metabolic recycling of the surplus energy to acetyl-CoA can increase squalene production in yeast, concomitant with minimizing the metabolic interferences in inherent pathways. Moreover, by integrating multiple copies of the rate-limiting enzyme and implementing N-degron-dependent protein degradation to downregulate the competing pathway, we systematically rewired the metabolic flux toward squalene, enabling remarkable squalene production (1024.88 mg/L in a shake flask). Ultimately, further optimization of the fed-batch fermentation process enabled remarkable squalene production of 6.53 g/L. Conclusions Our demonstration of squalene production via engineered yeast suggests that plant- or animal-based supplies of medicinal squalene can potentially be complemented or replaced by industrial fermentation. This approach will also provide a universal strategy for the more stable and sustainable production of high-value terpenes. |
| first_indexed | 2024-04-11T19:33:57Z |
| format | Article |
| id | doaj.art-362cca64b78e45f7bbfe25d4656374c9 |
| institution | Directory Open Access Journal |
| issn | 2731-3654 |
| language | English |
| last_indexed | 2024-04-11T19:33:57Z |
| publishDate | 2022-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj.art-362cca64b78e45f7bbfe25d4656374c92022-12-22T04:06:56ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542022-10-0115111110.1186/s13068-022-02208-9Metabolic recycling of storage lipids promotes squalene biosynthesis in yeastSo-Hee Son0Jae-Eung Kim1Soo Young Moon2In-Seung Jang3Byung Jo Yu4Ju Young Lee5Research Center for Bio-Based Chemistry, Korea Research Institute of Chemical Technology (KRICT)Research Center for Bio-Based Chemistry, Korea Research Institute of Chemical Technology (KRICT)Research Center for Bio-Based Chemistry, Korea Research Institute of Chemical Technology (KRICT)Intelligent Sustainable Materials R&D Group, Research Institute of Sustainable Manufacturing System, Korea Institute of Industrial Technology (KITECH)Intelligent Sustainable Materials R&D Group, Research Institute of Sustainable Manufacturing System, Korea Institute of Industrial Technology (KITECH)Research Center for Bio-Based Chemistry, Korea Research Institute of Chemical Technology (KRICT)Abstract Background Metabolic rewiring in microbes is an economical and sustainable strategy for synthesizing valuable natural terpenes. Terpenes are the largest class of nature-derived specialized metabolites, and many have valuable pharmaceutical or biological activity. Squalene, a medicinal terpene, is used as a vaccine adjuvant to improve the efficacy of vaccines, including pandemic coronavirus disease 2019 (COVID-19) vaccines, and plays diverse biological roles as an antioxidant and anticancer agent. However, metabolic rewiring interferes with inherent metabolic pathways, often in a way that impairs the cellular growth and fitness of the microbial host. In particular, as the key starting molecule for producing various compounds including squalene, acetyl-CoA is involved in numerous biological processes with tight regulation to maintain metabolic homeostasis, which limits redirection of metabolic fluxes toward desired products. Results In this study, focusing on the recycling of surplus metabolic energy stored in lipid droplets, we show that the metabolic recycling of the surplus energy to acetyl-CoA can increase squalene production in yeast, concomitant with minimizing the metabolic interferences in inherent pathways. Moreover, by integrating multiple copies of the rate-limiting enzyme and implementing N-degron-dependent protein degradation to downregulate the competing pathway, we systematically rewired the metabolic flux toward squalene, enabling remarkable squalene production (1024.88 mg/L in a shake flask). Ultimately, further optimization of the fed-batch fermentation process enabled remarkable squalene production of 6.53 g/L. Conclusions Our demonstration of squalene production via engineered yeast suggests that plant- or animal-based supplies of medicinal squalene can potentially be complemented or replaced by industrial fermentation. This approach will also provide a universal strategy for the more stable and sustainable production of high-value terpenes.https://doi.org/10.1186/s13068-022-02208-9Lipid dropletMetabolic recyclingYeastTerpeneSqualeneMetabolic engineering |
| spellingShingle | So-Hee Son Jae-Eung Kim Soo Young Moon In-Seung Jang Byung Jo Yu Ju Young Lee Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast Biotechnology for Biofuels and Bioproducts Lipid droplet Metabolic recycling Yeast Terpene Squalene Metabolic engineering |
| title | Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| title_full | Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| title_fullStr | Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| title_full_unstemmed | Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| title_short | Metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| title_sort | metabolic recycling of storage lipids promotes squalene biosynthesis in yeast |
| topic | Lipid droplet Metabolic recycling Yeast Terpene Squalene Metabolic engineering |
| url | https://doi.org/10.1186/s13068-022-02208-9 |
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