Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production
Carnosic acid (CA), a phenolic tricyclic diterpene, has many biological effects, including anti-inflammatory, anticancer, antiobesity, and antidiabetic activities. In this study, an efficient biosynthetic pathway was constructed to produce CA in Saccharomyces cerevisiae. First, the CA precursor milt...
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| Format: | Article |
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2022.916605/full |
| _version_ | 1828338182627786752 |
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| author | Panpan Wei Chuanbo Zhang Xueke Bian Wenyu Lu Wenyu Lu Wenyu Lu |
| author_facet | Panpan Wei Chuanbo Zhang Xueke Bian Wenyu Lu Wenyu Lu Wenyu Lu |
| author_sort | Panpan Wei |
| collection | DOAJ |
| description | Carnosic acid (CA), a phenolic tricyclic diterpene, has many biological effects, including anti-inflammatory, anticancer, antiobesity, and antidiabetic activities. In this study, an efficient biosynthetic pathway was constructed to produce CA in Saccharomyces cerevisiae. First, the CA precursor miltiradiene was synthesized, after which the CA production strain was constructed by integrating the genes encoding cytochrome P450 enzymes (P450s) and cytochrome P450 reductase (CPR) SmCPR. The CA titer was further increased by the coexpression of CYP76AH1 and SmCPR ∼t28SpCytb5 fusion proteins and the overexpression of different catalases to detoxify the hydrogen peroxide (H2O2). Finally, engineering of the endoplasmic reticulum and cofactor supply increased the CA titer to 24.65 mg/L in shake flasks and 75.18 mg/L in 5 L fed-batch fermentation. This study demonstrates that the ability of engineered yeast cells to synthesize CA can be improved through metabolic engineering and synthetic biology strategies, providing a theoretical basis for microbial synthesis of other diterpenoids. |
| first_indexed | 2024-04-13T22:24:16Z |
| format | Article |
| id | doaj.art-ee4b7d7bfe3047fcae1aec0d4b7f1c94 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-13T22:24:16Z |
| publishDate | 2022-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-ee4b7d7bfe3047fcae1aec0d4b7f1c942022-12-22T02:27:06ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852022-06-011010.3389/fbioe.2022.916605916605Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid ProductionPanpan Wei0Chuanbo Zhang1Xueke Bian2Wenyu Lu3Wenyu Lu4Wenyu Lu5School of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSchool of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSchool of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaSchool of Chemical Engineering and Technology, Tianjin University, Tianjin, ChinaKey Laboratory of Systems Bioengineering of the Ministry of Education, Tianjin University, Tianjin, ChinaSynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin, ChinaCarnosic acid (CA), a phenolic tricyclic diterpene, has many biological effects, including anti-inflammatory, anticancer, antiobesity, and antidiabetic activities. In this study, an efficient biosynthetic pathway was constructed to produce CA in Saccharomyces cerevisiae. First, the CA precursor miltiradiene was synthesized, after which the CA production strain was constructed by integrating the genes encoding cytochrome P450 enzymes (P450s) and cytochrome P450 reductase (CPR) SmCPR. The CA titer was further increased by the coexpression of CYP76AH1 and SmCPR ∼t28SpCytb5 fusion proteins and the overexpression of different catalases to detoxify the hydrogen peroxide (H2O2). Finally, engineering of the endoplasmic reticulum and cofactor supply increased the CA titer to 24.65 mg/L in shake flasks and 75.18 mg/L in 5 L fed-batch fermentation. This study demonstrates that the ability of engineered yeast cells to synthesize CA can be improved through metabolic engineering and synthetic biology strategies, providing a theoretical basis for microbial synthesis of other diterpenoids.https://www.frontiersin.org/articles/10.3389/fbioe.2022.916605/fullcarnosic acidSaccharomyces cerevisiaeterpenoidmiltiradienesynthetic biology |
| spellingShingle | Panpan Wei Chuanbo Zhang Xueke Bian Wenyu Lu Wenyu Lu Wenyu Lu Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production Frontiers in Bioengineering and Biotechnology carnosic acid Saccharomyces cerevisiae terpenoid miltiradiene synthetic biology |
| title | Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production |
| title_full | Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production |
| title_fullStr | Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production |
| title_full_unstemmed | Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production |
| title_short | Metabolic Engineering of Saccharomyces cerevisiae for Heterologous Carnosic Acid Production |
| title_sort | metabolic engineering of saccharomyces cerevisiae for heterologous carnosic acid production |
| topic | carnosic acid Saccharomyces cerevisiae terpenoid miltiradiene synthetic biology |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2022.916605/full |
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