Metabolic engineering of Rhodotorula toruloides for resveratrol production
Abstract Background Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial producti...
| 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/s12934-022-02006-w |
| _version_ | 1797977184847003648 |
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| author | Mengyao Zhang Qidou Gao Yijuan Liu Zhumei Fang Zhiwei Gong Zongbao K. Zhao Xiaobing Yang |
| author_facet | Mengyao Zhang Qidou Gao Yijuan Liu Zhumei Fang Zhiwei Gong Zongbao K. Zhao Xiaobing Yang |
| author_sort | Mengyao Zhang |
| collection | DOAJ |
| description | Abstract Background Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of resveratrol offers a promising alternative to plant- and chemical-based processes. The non-conventional oleaginous yeast Rhodotorula toruloides is a potential workhorse for the production of resveratrol that endowed with an efficient and intrinsic bifunctional phenylalanine/tyrosine ammonia-lyase (RtPAL) and malonyl-CoA pool, which may facilitate the resveratrol synthesis when properly rewired. Results Resveratrol showed substantial stability and would not affect the R. toruloides growth during the yeast cultivation in flasks. The heterologus resveratrol biosynthesis pathway was established by introducing the 4-coumaroyl-CoA ligase (At4CL), and the stilbene synthase (VlSTS) from Arabidopsis thaliana and Vitis labrusca, respectively. Next, The resveratrol production was increased by 634% through employing the cinnamate-4-hydroxylase from A. thaliana (AtC4H), the fused protein At4CL::VlSTS, the cytochrome P450 reductase 2 from A. thaliana (AtATR2) and the endogenous cytochrome B5 of R. toruloides (RtCYB5). Then, the related endogenous pathways were optimized to affect a further 60% increase. Finally, the engineered strain produced a maximum titer of 125.2 mg/L resveratrol in YPD medium. Conclusion The non-conventional oleaginous yeast R. toruloides was engineered for the first time to produce resveratrol. Protein fusion, co-factor channeling, and ARO4 and ARO7 overexpression were efficient for improving resveratrol production. The results demonstrated the potential of R. toruloides for resveratrol and other phenylpropanoids production. |
| first_indexed | 2024-04-11T05:02:55Z |
| format | Article |
| id | doaj.art-394de580279e4fec8da14e8af749af11 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-11T05:02:55Z |
| publishDate | 2022-12-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-394de580279e4fec8da14e8af749af112022-12-25T12:34:47ZengBMCMicrobial Cell Factories1475-28592022-12-012111910.1186/s12934-022-02006-wMetabolic engineering of Rhodotorula toruloides for resveratrol productionMengyao Zhang0Qidou Gao1Yijuan Liu2Zhumei Fang3Zhiwei Gong4Zongbao K. Zhao5Xiaobing Yang6College of Enology, Northwest A&F UniversityCollege of Enology, Northwest A&F UniversityCollege of Enology, Northwest A&F UniversityCollege of Enology, Northwest A&F UniversitySchool of Chemistry and Chemical Engineering, Wuhan University of Science and TechnologyDivision of Biotechnology, Dalian Institute of Chemical Physics, Chinese Academy of SciencesCollege of Enology, Northwest A&F UniversityAbstract Background Resveratrol is a plant-derived phenylpropanoid with diverse biological activities and pharmacological applications. Plant-based extraction could not satisfy ever-increasing market demand, while chemical synthesis is impeded by the existence of toxic impurities. Microbial production of resveratrol offers a promising alternative to plant- and chemical-based processes. The non-conventional oleaginous yeast Rhodotorula toruloides is a potential workhorse for the production of resveratrol that endowed with an efficient and intrinsic bifunctional phenylalanine/tyrosine ammonia-lyase (RtPAL) and malonyl-CoA pool, which may facilitate the resveratrol synthesis when properly rewired. Results Resveratrol showed substantial stability and would not affect the R. toruloides growth during the yeast cultivation in flasks. The heterologus resveratrol biosynthesis pathway was established by introducing the 4-coumaroyl-CoA ligase (At4CL), and the stilbene synthase (VlSTS) from Arabidopsis thaliana and Vitis labrusca, respectively. Next, The resveratrol production was increased by 634% through employing the cinnamate-4-hydroxylase from A. thaliana (AtC4H), the fused protein At4CL::VlSTS, the cytochrome P450 reductase 2 from A. thaliana (AtATR2) and the endogenous cytochrome B5 of R. toruloides (RtCYB5). Then, the related endogenous pathways were optimized to affect a further 60% increase. Finally, the engineered strain produced a maximum titer of 125.2 mg/L resveratrol in YPD medium. Conclusion The non-conventional oleaginous yeast R. toruloides was engineered for the first time to produce resveratrol. Protein fusion, co-factor channeling, and ARO4 and ARO7 overexpression were efficient for improving resveratrol production. The results demonstrated the potential of R. toruloides for resveratrol and other phenylpropanoids production.https://doi.org/10.1186/s12934-022-02006-wRhodotorula toruloidesResveratrolMetabolic engineeringCell factory |
| spellingShingle | Mengyao Zhang Qidou Gao Yijuan Liu Zhumei Fang Zhiwei Gong Zongbao K. Zhao Xiaobing Yang Metabolic engineering of Rhodotorula toruloides for resveratrol production Microbial Cell Factories Rhodotorula toruloides Resveratrol Metabolic engineering Cell factory |
| title | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
| title_full | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
| title_fullStr | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
| title_full_unstemmed | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
| title_short | Metabolic engineering of Rhodotorula toruloides for resveratrol production |
| title_sort | metabolic engineering of rhodotorula toruloides for resveratrol production |
| topic | Rhodotorula toruloides Resveratrol Metabolic engineering Cell factory |
| url | https://doi.org/10.1186/s12934-022-02006-w |
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