The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma
Shikonin derivatives are natural naphthoquinone compounds and the main bioactive components produced by several boraginaceous plants, such as Lithospermum erythrorhizon and Arnebia euchroma. Phytochemical studies utilizing both L. erythrorhizon and A. euchroma cultured cells indicate the existence o...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-04-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2023.1160571/full |
| _version_ | 1827960394494246912 |
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| author | Ruishan Wang Changzheng Liu Chaogeng Lyu Jiahui Sun Chuanzhi Kang Ying Ma Xiufu Wan Juan Guo Linyuan Shi Jinye Wang Luqi Huang Sheng Wang Lanping Guo |
| author_facet | Ruishan Wang Changzheng Liu Chaogeng Lyu Jiahui Sun Chuanzhi Kang Ying Ma Xiufu Wan Juan Guo Linyuan Shi Jinye Wang Luqi Huang Sheng Wang Lanping Guo |
| author_sort | Ruishan Wang |
| collection | DOAJ |
| description | Shikonin derivatives are natural naphthoquinone compounds and the main bioactive components produced by several boraginaceous plants, such as Lithospermum erythrorhizon and Arnebia euchroma. Phytochemical studies utilizing both L. erythrorhizon and A. euchroma cultured cells indicate the existence of a competing route branching out from the shikonin biosynthetic pathway to shikonofuran. A previous study has shown that the branch point is the transformation from (Z)-3’’-hydroxy-geranylhydroquinone to an aldehyde intermediate (E)-3’’-oxo-geranylhydroquinone. However, the gene encoding the oxidoreductase that catalyzes the branch reaction remains unidentified. In this study, we discovered a candidate gene belonging to the cinnamyl alcohol dehydrogenase family, AeHGO, through coexpression analysis of transcriptome data sets of shikonin-proficient and shikonin-deficient cell lines of A. euchroma. In biochemical assays, purified AeHGO protein reversibly oxidized (Z)-3’’-hydroxy-geranylhydroquinone to produce (E)-3’’-oxo-geranylhydroquinone followed by reversibly reducing (E)-3’’-oxo-geranylhydroquinone to (E)-3’’-hydroxy-geranylhydroquinone, resulting in an equilibrium mixture of the three compounds. Time course analysis and kinetic parameters showed that the reduction of (E)-3’’-oxo-geranylhydroquinone was stereoselective and efficient in presence of NADPH, which determined that the overall reaction proceeded from (Z)-3’’-hydroxy-geranylhydroquinone to (E)-3’’-hydroxy-geranylhydroquinone. Considering that there is a competition between the accumulation of shikonin and shikonofuran derivatives in cultured plant cells, AeHGO is supposed to play an important role in the metabolic regulation of the shikonin biosynthetic pathway. Characterization of AeHGO should help expedite the development of metabolic engineering and synthetic biology toward production of shikonin derivatives. |
| first_indexed | 2024-04-09T16:06:37Z |
| format | Article |
| id | doaj.art-a59dc5ffcb604729a11c252556b300fd |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-09T16:06:37Z |
| publishDate | 2023-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-a59dc5ffcb604729a11c252556b300fd2023-04-25T05:21:16ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2023-04-011410.3389/fpls.2023.11605711160571The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchromaRuishan WangChangzheng LiuChaogeng LyuJiahui SunChuanzhi KangYing MaXiufu WanJuan GuoLinyuan ShiJinye WangLuqi HuangSheng WangLanping GuoShikonin derivatives are natural naphthoquinone compounds and the main bioactive components produced by several boraginaceous plants, such as Lithospermum erythrorhizon and Arnebia euchroma. Phytochemical studies utilizing both L. erythrorhizon and A. euchroma cultured cells indicate the existence of a competing route branching out from the shikonin biosynthetic pathway to shikonofuran. A previous study has shown that the branch point is the transformation from (Z)-3’’-hydroxy-geranylhydroquinone to an aldehyde intermediate (E)-3’’-oxo-geranylhydroquinone. However, the gene encoding the oxidoreductase that catalyzes the branch reaction remains unidentified. In this study, we discovered a candidate gene belonging to the cinnamyl alcohol dehydrogenase family, AeHGO, through coexpression analysis of transcriptome data sets of shikonin-proficient and shikonin-deficient cell lines of A. euchroma. In biochemical assays, purified AeHGO protein reversibly oxidized (Z)-3’’-hydroxy-geranylhydroquinone to produce (E)-3’’-oxo-geranylhydroquinone followed by reversibly reducing (E)-3’’-oxo-geranylhydroquinone to (E)-3’’-hydroxy-geranylhydroquinone, resulting in an equilibrium mixture of the three compounds. Time course analysis and kinetic parameters showed that the reduction of (E)-3’’-oxo-geranylhydroquinone was stereoselective and efficient in presence of NADPH, which determined that the overall reaction proceeded from (Z)-3’’-hydroxy-geranylhydroquinone to (E)-3’’-hydroxy-geranylhydroquinone. Considering that there is a competition between the accumulation of shikonin and shikonofuran derivatives in cultured plant cells, AeHGO is supposed to play an important role in the metabolic regulation of the shikonin biosynthetic pathway. Characterization of AeHGO should help expedite the development of metabolic engineering and synthetic biology toward production of shikonin derivatives.https://www.frontiersin.org/articles/10.3389/fpls.2023.1160571/fullshikonin derivativesshikonofuran derivativesArnebia euchromabiosynthesiscinnamyl alcohol dehydrogenaseAeHGO |
| spellingShingle | Ruishan Wang Changzheng Liu Chaogeng Lyu Jiahui Sun Chuanzhi Kang Ying Ma Xiufu Wan Juan Guo Linyuan Shi Jinye Wang Luqi Huang Sheng Wang Lanping Guo The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma Frontiers in Plant Science shikonin derivatives shikonofuran derivatives Arnebia euchroma biosynthesis cinnamyl alcohol dehydrogenase AeHGO |
| title | The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma |
| title_full | The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma |
| title_fullStr | The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma |
| title_full_unstemmed | The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma |
| title_short | The discovery and characterization of AeHGO in the branching route from shikonin biosynthesis to shikonofuran in Arnebia euchroma |
| title_sort | discovery and characterization of aehgo in the branching route from shikonin biosynthesis to shikonofuran in arnebia euchroma |
| topic | shikonin derivatives shikonofuran derivatives Arnebia euchroma biosynthesis cinnamyl alcohol dehydrogenase AeHGO |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2023.1160571/full |
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