Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica
β-carotene is a precursor of vitamin A and has multiple physiological functions. Producing β-carotene by microbial fermentation has attracted much attention to consumers’ preference for natural products. This study focused on improving β-carotene production by constructing codon-adapted genes and mi...
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Frontiers Media S.A.
2021-03-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.627150/full |
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| author | Liang Liu Yu Ling Qu Gui Ru Dong Jing Wang Ching Yuan Hu Ching Yuan Hu Yong Hong Meng |
| author_facet | Liang Liu Yu Ling Qu Gui Ru Dong Jing Wang Ching Yuan Hu Ching Yuan Hu Yong Hong Meng |
| author_sort | Liang Liu |
| collection | DOAJ |
| description | β-carotene is a precursor of vitamin A and has multiple physiological functions. Producing β-carotene by microbial fermentation has attracted much attention to consumers’ preference for natural products. This study focused on improving β-carotene production by constructing codon-adapted genes and minimizing intermediate accumulation. The codon-adapted CarRA and CarB genes from the industrial strain of Blakeslea trispora were integrated into the genome of the Yarrowia lipolytica to construct YL-C0, the baseline strain for producing β-carotene. Thereafter, the β-carotene biosynthetic pathway’s metabolic balance was accurately regulated to reduce the intermediates’ accumulation. Notably, the β-carotene content increased by 21 times to reach 12.5 dry cell weight (DCW) mg/g when minimizing HMG-CoA and FPP accumulation. Further, we improved the expression levels of the CarRA and CarB genes to minimize the accumulation of phytoene and lycopene. Total production of β-carotene of 1.7 g/L and 21.6 mg/g DCW was achieved. These results reveal that the rate-limiting enzymes CarRA and CarB of B. trispora exhibited higher catalytic activity than the same enzymes from other microorganisms. Promoting metabolic balance by minimizing the accumulation of intermediates is a very effective strategy for increasing β-carotene. The β-carotene-producing strain constructed in this study has established the foundation for its potential use in industrial production. These successful engineering strategies also provide a foundation for large-scale production of other terpenoids. |
| first_indexed | 2024-12-16T17:07:37Z |
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| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-16T17:07:37Z |
| publishDate | 2021-03-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-3cbecb3fd6444a2eae6000a3dd9f19fd2022-12-21T22:23:31ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-03-011210.3389/fmicb.2021.627150627150Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolyticaLiang Liu0Yu Ling Qu1Gui Ru Dong2Jing Wang3Ching Yuan Hu4Ching Yuan Hu5Yong Hong Meng6Engineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, ChinaEngineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, ChinaEngineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, ChinaEngineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, ChinaEngineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, ChinaDepartment of Human Nutrition, Food and Animal Sciences, College of Tropical Agriculture and Human Resources, University of Hawai’i at Mānoa, Honolulu, HI, United StatesEngineering Research Center of High Value Utilization of Western China Fruit Resources, Ministry of Education, National Research and Development Center of Apple Processing Technology, Shaanxi Engineering Laboratory for Food Green Processing and Safety Control, College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an, Chinaβ-carotene is a precursor of vitamin A and has multiple physiological functions. Producing β-carotene by microbial fermentation has attracted much attention to consumers’ preference for natural products. This study focused on improving β-carotene production by constructing codon-adapted genes and minimizing intermediate accumulation. The codon-adapted CarRA and CarB genes from the industrial strain of Blakeslea trispora were integrated into the genome of the Yarrowia lipolytica to construct YL-C0, the baseline strain for producing β-carotene. Thereafter, the β-carotene biosynthetic pathway’s metabolic balance was accurately regulated to reduce the intermediates’ accumulation. Notably, the β-carotene content increased by 21 times to reach 12.5 dry cell weight (DCW) mg/g when minimizing HMG-CoA and FPP accumulation. Further, we improved the expression levels of the CarRA and CarB genes to minimize the accumulation of phytoene and lycopene. Total production of β-carotene of 1.7 g/L and 21.6 mg/g DCW was achieved. These results reveal that the rate-limiting enzymes CarRA and CarB of B. trispora exhibited higher catalytic activity than the same enzymes from other microorganisms. Promoting metabolic balance by minimizing the accumulation of intermediates is a very effective strategy for increasing β-carotene. The β-carotene-producing strain constructed in this study has established the foundation for its potential use in industrial production. These successful engineering strategies also provide a foundation for large-scale production of other terpenoids.https://www.frontiersin.org/articles/10.3389/fmicb.2021.627150/fullβ-carotenemetabolic balanceCarRAYarrowia lipolyticaCarB |
| spellingShingle | Liang Liu Yu Ling Qu Gui Ru Dong Jing Wang Ching Yuan Hu Ching Yuan Hu Yong Hong Meng Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica Frontiers in Microbiology β-carotene metabolic balance CarRA Yarrowia lipolytica CarB |
| title | Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica |
| title_full | Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica |
| title_fullStr | Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica |
| title_full_unstemmed | Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica |
| title_short | Elevated β-Carotene Production Using Codon-Adapted CarRA&B and Metabolic Balance in Engineered Yarrowia lipolytica |
| title_sort | elevated β carotene production using codon adapted carra b and metabolic balance in engineered yarrowia lipolytica |
| topic | β-carotene metabolic balance CarRA Yarrowia lipolytica CarB |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.627150/full |
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