Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction
Abstract Background α-Humulene is a plant-derived monocyclic sesquiterpenoid with multiple pharmacological activities, and far-reaching potential for the development of new drugs. Currently, the production of α-humulene is typically achieved via plant extraction, which is not sustainable and limited...
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| Format: | Article |
| Language: | English |
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BMC
2022-05-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
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| Online Access: | https://doi.org/10.1186/s13068-022-02160-8 |
| _version_ | 1828796391264092160 |
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| author | Lihua Zhang Haiquan Yang Yuanyuan Xia Wei Shen Liming Liu Qi Li Xianzhong Chen |
| author_facet | Lihua Zhang Haiquan Yang Yuanyuan Xia Wei Shen Liming Liu Qi Li Xianzhong Chen |
| author_sort | Lihua Zhang |
| collection | DOAJ |
| description | Abstract Background α-Humulene is a plant-derived monocyclic sesquiterpenoid with multiple pharmacological activities, and far-reaching potential for the development of new drugs. Currently, the production of α-humulene is typically achieved via plant extraction, which is not sustainable and limited by low yields. The oleaginous yeast Candida tropicalis has recently emerged as a valuable host for producing high-value-added chemicals. However, the potential of C. tropicalis for terpenoid production has not been exploited. Results In this study, C. tropicalis was engineered for de novo synthesis of α-humulene from glucose. To improve α-humulene production, the codon-optimised α-humulene synthase gene and the entire endogenous farnesyl diphosphate synthesis pathway were co-overexpressed. Furthermore, bottlenecks in the α-humulene synthase pathway were identified and relieved by overexpressing α-humulene synthase, acetoacetyl-CoA thiolase and NADH-dependent HMG-CoA reductase. Combined with fermentation medium optimisation, the engineered strain produced 195.31 mg/L of α-humulene in shake flasks and 4115.42 mg/L in a bioreactor through fed-batch fermentation, a 253- and 5345-fold increase over the initial production, respectively. Conclusions This study demonstrates the potential of C. tropicalis for α-humulene production, and presents a platform for the biosynthesis of other terpenoids. |
| first_indexed | 2024-12-12T04:21:58Z |
| format | Article |
| id | doaj.art-fbc5091ac88e45d5a9527d6332a65822 |
| institution | Directory Open Access Journal |
| issn | 2731-3654 |
| language | English |
| last_indexed | 2024-12-12T04:21:58Z |
| publishDate | 2022-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj.art-fbc5091ac88e45d5a9527d6332a658222022-12-22T00:38:18ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542022-05-0115111210.1186/s13068-022-02160-8Engineering the oleaginous yeast Candida tropicalis for α-humulene overproductionLihua Zhang0Haiquan Yang1Yuanyuan Xia2Wei Shen3Liming Liu4Qi Li5Xianzhong Chen6Key Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityKey Laboratory of Industrial Biotechnology, Ministry of Education, and School of Biotechnology, School of Biotechnology, Jiangnan UniversityAbstract Background α-Humulene is a plant-derived monocyclic sesquiterpenoid with multiple pharmacological activities, and far-reaching potential for the development of new drugs. Currently, the production of α-humulene is typically achieved via plant extraction, which is not sustainable and limited by low yields. The oleaginous yeast Candida tropicalis has recently emerged as a valuable host for producing high-value-added chemicals. However, the potential of C. tropicalis for terpenoid production has not been exploited. Results In this study, C. tropicalis was engineered for de novo synthesis of α-humulene from glucose. To improve α-humulene production, the codon-optimised α-humulene synthase gene and the entire endogenous farnesyl diphosphate synthesis pathway were co-overexpressed. Furthermore, bottlenecks in the α-humulene synthase pathway were identified and relieved by overexpressing α-humulene synthase, acetoacetyl-CoA thiolase and NADH-dependent HMG-CoA reductase. Combined with fermentation medium optimisation, the engineered strain produced 195.31 mg/L of α-humulene in shake flasks and 4115.42 mg/L in a bioreactor through fed-batch fermentation, a 253- and 5345-fold increase over the initial production, respectively. Conclusions This study demonstrates the potential of C. tropicalis for α-humulene production, and presents a platform for the biosynthesis of other terpenoids.https://doi.org/10.1186/s13068-022-02160-8Candida tropicalisα-HumuleneRate-limiting enzymesMetabolic engineeringMevalonate pathway |
| spellingShingle | Lihua Zhang Haiquan Yang Yuanyuan Xia Wei Shen Liming Liu Qi Li Xianzhong Chen Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction Biotechnology for Biofuels and Bioproducts Candida tropicalis α-Humulene Rate-limiting enzymes Metabolic engineering Mevalonate pathway |
| title | Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction |
| title_full | Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction |
| title_fullStr | Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction |
| title_full_unstemmed | Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction |
| title_short | Engineering the oleaginous yeast Candida tropicalis for α-humulene overproduction |
| title_sort | engineering the oleaginous yeast candida tropicalis for α humulene overproduction |
| topic | Candida tropicalis α-Humulene Rate-limiting enzymes Metabolic engineering Mevalonate pathway |
| url | https://doi.org/10.1186/s13068-022-02160-8 |
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