Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering
Longifolene (C15H24) is a tricyclic sesquiterpene widely utilized in the cosmetics and fragrances due to its versatile applications. Traditional extraction methods from plants suffer from low titer and lengthy production cycles, while chemical synthesis is hampered by the compound's complex str...
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| Format: | Article |
| Language: | English |
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KeAi Communications Co., Ltd.
2025-06-01
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| Series: | Synthetic and Systems Biotechnology |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405805X25000043 |
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| author | Yi-Tong Yao Xiao Zhang Chen-Yu Wang Yu-He Zhang Da-Wei Li Wei-Dong Yang Hong-Ye Li Li-Gong Zou |
| author_facet | Yi-Tong Yao Xiao Zhang Chen-Yu Wang Yu-He Zhang Da-Wei Li Wei-Dong Yang Hong-Ye Li Li-Gong Zou |
| author_sort | Yi-Tong Yao |
| collection | DOAJ |
| description | Longifolene (C15H24) is a tricyclic sesquiterpene widely utilized in the cosmetics and fragrances due to its versatile applications. Traditional extraction methods from plants suffer from low titer and lengthy production cycles, while chemical synthesis is hampered by the compound's complex structure, leading to high costs and insufficient market supply. This study aimed to develop a microbial cell factory for enhanced longifolene production. The strategy involved integrating longifolene synthase from Pinus sylvestris (PsTPS) into Yarrowia lipolytica and employing multiple metabolic engineering approaches. Initially, key genes in the mevalonate (MVA) pathway were overexpressed to enhance longifolene precursor availability for longifolene biosynthesis. Subsequently, protein engineering techniques were applied to optimize PsTPS (tPsTPS) for improved catalytic efficiency. Furthermore, co-expression of molecular chaperones was implemented to enhance the synthesis and secretion of PsTPS. The introduction of the isopentenol utilization pathway (IUP) further augmented the supply of C5 substrate. By optimizing the culture conditions, including a reduction in culture temperature, the efflux of longifolene was increased, and the dissolved oxygen levels were enhanced to promote the growth of the strain. These collective efforts resulted culminated in the engineered strain Z03 achieving a noteworthy production level of 34.67 mg/L of longifolene in shake flasks. This study not only demonstrates the feasibility of enhancing sesquiterpene production in Y. lipolytica but also highlights the potential of microbial platforms in meeting industrial demands for complex natural products. |
| format | Article |
| id | doaj-art-94c4b81def5a47faaf7a8d3c6dd747ce |
| institution | Kabale University |
| issn | 2405-805X |
| language | English |
| publishDate | 2025-06-01 |
| publisher | KeAi Communications Co., Ltd. |
| record_format | Article |
| series | Synthetic and Systems Biotechnology |
| spelling | doaj-art-94c4b81def5a47faaf7a8d3c6dd747ce2025-01-26T05:04:13ZengKeAi Communications Co., Ltd.Synthetic and Systems Biotechnology2405-805X2025-06-01102433441Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineeringYi-Tong Yao0Xiao Zhang1Chen-Yu Wang2Yu-He Zhang3Da-Wei Li4Wei-Dong Yang5Hong-Ye Li6Li-Gong Zou7College of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCollege of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaCorresponding author.; College of Life Science and Technology, Jinan University, Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Guangzhou, 510632, ChinaLongifolene (C15H24) is a tricyclic sesquiterpene widely utilized in the cosmetics and fragrances due to its versatile applications. Traditional extraction methods from plants suffer from low titer and lengthy production cycles, while chemical synthesis is hampered by the compound's complex structure, leading to high costs and insufficient market supply. This study aimed to develop a microbial cell factory for enhanced longifolene production. The strategy involved integrating longifolene synthase from Pinus sylvestris (PsTPS) into Yarrowia lipolytica and employing multiple metabolic engineering approaches. Initially, key genes in the mevalonate (MVA) pathway were overexpressed to enhance longifolene precursor availability for longifolene biosynthesis. Subsequently, protein engineering techniques were applied to optimize PsTPS (tPsTPS) for improved catalytic efficiency. Furthermore, co-expression of molecular chaperones was implemented to enhance the synthesis and secretion of PsTPS. The introduction of the isopentenol utilization pathway (IUP) further augmented the supply of C5 substrate. By optimizing the culture conditions, including a reduction in culture temperature, the efflux of longifolene was increased, and the dissolved oxygen levels were enhanced to promote the growth of the strain. These collective efforts resulted culminated in the engineered strain Z03 achieving a noteworthy production level of 34.67 mg/L of longifolene in shake flasks. This study not only demonstrates the feasibility of enhancing sesquiterpene production in Y. lipolytica but also highlights the potential of microbial platforms in meeting industrial demands for complex natural products.http://www.sciencedirect.com/science/article/pii/S2405805X25000043LongifoleneYarrowia lipolyticaSesquiterpeneMolecular chaperoneProtein engineeringIsopentenol utilization pathway |
| spellingShingle | Yi-Tong Yao Xiao Zhang Chen-Yu Wang Yu-He Zhang Da-Wei Li Wei-Dong Yang Hong-Ye Li Li-Gong Zou Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering Synthetic and Systems Biotechnology Longifolene Yarrowia lipolytica Sesquiterpene Molecular chaperone Protein engineering Isopentenol utilization pathway |
| title | Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering |
| title_full | Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering |
| title_fullStr | Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering |
| title_full_unstemmed | Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering |
| title_short | Optimizing longifolene production in Yarrowia lipolytica via metabolic and protein engineering |
| title_sort | optimizing longifolene production in yarrowia lipolytica via metabolic and protein engineering |
| topic | Longifolene Yarrowia lipolytica Sesquiterpene Molecular chaperone Protein engineering Isopentenol utilization pathway |
| url | http://www.sciencedirect.com/science/article/pii/S2405805X25000043 |
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