Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production
Lutein, made by the α-branch of the methyl-erythritol phosphate (MEP) pathway, is one of the most abundant xanthophylls in plants. It is involved in the structural stabilization of light-harvesting complexes, transfer of excitation energy to chlorophylls and photoprotection. In contrast, lutein and...
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Frontiers Media S.A.
2021-07-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2021.699424/full |
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| author | Martin Lehmann Evgenia Vamvaka Alejandro Torrado Peter Jahns Marcel Dann Lea Rosenhammer Amel Aziba Dario Leister Thilo Rühle |
| author_facet | Martin Lehmann Evgenia Vamvaka Alejandro Torrado Peter Jahns Marcel Dann Lea Rosenhammer Amel Aziba Dario Leister Thilo Rühle |
| author_sort | Martin Lehmann |
| collection | DOAJ |
| description | Lutein, made by the α-branch of the methyl-erythritol phosphate (MEP) pathway, is one of the most abundant xanthophylls in plants. It is involved in the structural stabilization of light-harvesting complexes, transfer of excitation energy to chlorophylls and photoprotection. In contrast, lutein and the α-branch of the MEP pathway are not present in cyanobacteria. In this study, we genetically engineered the cyanobacterium Synechocystis for the missing MEP α-branch resulting in lutein accumulation. A cassette comprising four Arabidopsis thaliana genes coding for two lycopene cyclases (AtLCYe and AtLCYb) and two hydroxylases (AtCYP97A and AtCYP97C) was introduced into a Synechocystis strain that lacks the endogenous, cyanobacterial lycopene cyclase cruA. The resulting synlut strain showed wild-type growth and only moderate changes in total pigment composition under mixotrophic conditions, indicating that the cruA deficiency can be complemented by Arabidopsis lycopene cyclases leaving the endogenous β-branch intact. A combination of liquid chromatography, UV-Vis detection and mass spectrometry confirmed a low but distinct synthesis of lutein at rates of 4.8 ± 1.5 nmol per liter culture at OD730 (1.03 ± 0.47 mmol mol–1 chlorophyll). In conclusion, synlut provides a suitable platform to study the α-branch of the plastidic MEP pathway and other functions related to lutein in a cyanobacterial host system. |
| first_indexed | 2024-12-16T20:36:50Z |
| format | Article |
| id | doaj.art-1512f505d4104121a443e55a599fe318 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-12-16T20:36:50Z |
| publishDate | 2021-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-1512f505d4104121a443e55a599fe3182022-12-21T22:17:13ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2021-07-011210.3389/fpls.2021.699424699424Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein ProductionMartin Lehmann0Evgenia Vamvaka1Alejandro Torrado2Peter Jahns3Marcel Dann4Lea Rosenhammer5Amel Aziba6Dario Leister7Thilo Rühle8Plant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Biochemistry, Heinrich-Heine-University Düsseldorf, Düsseldorf, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyPlant Molecular Biology, Faculty of Biology, Ludwig-Maximilians-University Munich, Munich, GermanyLutein, made by the α-branch of the methyl-erythritol phosphate (MEP) pathway, is one of the most abundant xanthophylls in plants. It is involved in the structural stabilization of light-harvesting complexes, transfer of excitation energy to chlorophylls and photoprotection. In contrast, lutein and the α-branch of the MEP pathway are not present in cyanobacteria. In this study, we genetically engineered the cyanobacterium Synechocystis for the missing MEP α-branch resulting in lutein accumulation. A cassette comprising four Arabidopsis thaliana genes coding for two lycopene cyclases (AtLCYe and AtLCYb) and two hydroxylases (AtCYP97A and AtCYP97C) was introduced into a Synechocystis strain that lacks the endogenous, cyanobacterial lycopene cyclase cruA. The resulting synlut strain showed wild-type growth and only moderate changes in total pigment composition under mixotrophic conditions, indicating that the cruA deficiency can be complemented by Arabidopsis lycopene cyclases leaving the endogenous β-branch intact. A combination of liquid chromatography, UV-Vis detection and mass spectrometry confirmed a low but distinct synthesis of lutein at rates of 4.8 ± 1.5 nmol per liter culture at OD730 (1.03 ± 0.47 mmol mol–1 chlorophyll). In conclusion, synlut provides a suitable platform to study the α-branch of the plastidic MEP pathway and other functions related to lutein in a cyanobacterial host system.https://www.frontiersin.org/articles/10.3389/fpls.2021.699424/fullluteincarotenoidscyanobacteriaSynechocystiscyclasegenetic engineering |
| spellingShingle | Martin Lehmann Evgenia Vamvaka Alejandro Torrado Peter Jahns Marcel Dann Lea Rosenhammer Amel Aziba Dario Leister Thilo Rühle Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production Frontiers in Plant Science lutein carotenoids cyanobacteria Synechocystis cyclase genetic engineering |
| title | Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production |
| title_full | Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production |
| title_fullStr | Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production |
| title_full_unstemmed | Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production |
| title_short | Introduction of the Carotenoid Biosynthesis α-Branch Into Synechocystis sp. PCC 6803 for Lutein Production |
| title_sort | introduction of the carotenoid biosynthesis α branch into synechocystis sp pcc 6803 for lutein production |
| topic | lutein carotenoids cyanobacteria Synechocystis cyclase genetic engineering |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2021.699424/full |
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