Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis
Abstract Amorphadiene is the precursor to synthesize the antimalarial drug artemisinin. The production of amorphadiene and artemisinin from metabolically engineered microbes may provide an alternate to plant secondary metabolite extraction. Microbial consortia can offer division of labor, and microb...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2022-12-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-022-02010-0 |
| _version_ | 1797973401671827456 |
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| author | Monireh Marsafari Fidelis Azi Shaohua Dou Peng Xu |
| author_facet | Monireh Marsafari Fidelis Azi Shaohua Dou Peng Xu |
| author_sort | Monireh Marsafari |
| collection | DOAJ |
| description | Abstract Amorphadiene is the precursor to synthesize the antimalarial drug artemisinin. The production of amorphadiene and artemisinin from metabolically engineered microbes may provide an alternate to plant secondary metabolite extraction. Microbial consortia can offer division of labor, and microbial co-culture system can be leveraged to achieve cost-efficient production of natural products. Using a co-culture system of Y. lipolytica Po1f and Po1g strains, subcellular localization of ADS gene (encoding amorphadiene synthase) into the endoplasmic reticulum, co-utilization of mixed carbon source, and enlargement of the endoplasmic reticulum (ER) surface area, we were able to significantly improve amorphadiene production in this work. Using Po1g/PPtM and Po1f/AaADSERx3/iGFMPDU strains and co-utilization of 5 µM sodium acetate with 20 g/L glucose in YPD media, amorphadiene titer were increased to 65.094 mg/L. The enlargement of the ER surface area caused by the deletion of the PAH1 gene provided more subcellular ER space for the action of the ADS-tagged gene. It further increased the amorphadiene production to 71.74 mg/L. The results demonstrated that the importance of the spatial localization of critical enzymes, and manipulating metabolic flux in the co-culture of Y. lipolytica can be efficient over a single culture for the bioproduction of isoprenoid-related secondary metabolites in a modular manner. Graphical Abstract |
| first_indexed | 2024-04-11T04:03:42Z |
| format | Article |
| id | doaj.art-15baa7fbe8434f4ba5f90e3ef443b035 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-11T04:03:42Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-15baa7fbe8434f4ba5f90e3ef443b0352023-01-01T12:30:40ZengBMCMicrobial Cell Factories1475-28592022-12-0121111010.1186/s12934-022-02010-0Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesisMonireh Marsafari0Fidelis Azi1Shaohua Dou2Peng Xu3Department of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore CountyDepartment of Chemical Engineering, Guangdong Provincial Key Laboratory of Materials and Technologies for Energy Conversion (MATEC), Guangdong Technion – Israel Institute of TechnologyCollege of Life and Health, Dalian UniversityDepartment of Chemical, Biochemical and Environmental Engineering, University of Maryland Baltimore CountyAbstract Amorphadiene is the precursor to synthesize the antimalarial drug artemisinin. The production of amorphadiene and artemisinin from metabolically engineered microbes may provide an alternate to plant secondary metabolite extraction. Microbial consortia can offer division of labor, and microbial co-culture system can be leveraged to achieve cost-efficient production of natural products. Using a co-culture system of Y. lipolytica Po1f and Po1g strains, subcellular localization of ADS gene (encoding amorphadiene synthase) into the endoplasmic reticulum, co-utilization of mixed carbon source, and enlargement of the endoplasmic reticulum (ER) surface area, we were able to significantly improve amorphadiene production in this work. Using Po1g/PPtM and Po1f/AaADSERx3/iGFMPDU strains and co-utilization of 5 µM sodium acetate with 20 g/L glucose in YPD media, amorphadiene titer were increased to 65.094 mg/L. The enlargement of the ER surface area caused by the deletion of the PAH1 gene provided more subcellular ER space for the action of the ADS-tagged gene. It further increased the amorphadiene production to 71.74 mg/L. The results demonstrated that the importance of the spatial localization of critical enzymes, and manipulating metabolic flux in the co-culture of Y. lipolytica can be efficient over a single culture for the bioproduction of isoprenoid-related secondary metabolites in a modular manner. Graphical Abstracthttps://doi.org/10.1186/s12934-022-02010-0Y. lipolyticaCo-cultureAmorphadieneEndoplasmic reticulumCellular localization |
| spellingShingle | Monireh Marsafari Fidelis Azi Shaohua Dou Peng Xu Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis Microbial Cell Factories Y. lipolytica Co-culture Amorphadiene Endoplasmic reticulum Cellular localization |
| title | Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis |
| title_full | Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis |
| title_fullStr | Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis |
| title_full_unstemmed | Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis |
| title_short | Modular co-culture engineering of Yarrowia lipolytica for amorphadiene biosynthesis |
| title_sort | modular co culture engineering of yarrowia lipolytica for amorphadiene biosynthesis |
| topic | Y. lipolytica Co-culture Amorphadiene Endoplasmic reticulum Cellular localization |
| url | https://doi.org/10.1186/s12934-022-02010-0 |
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