Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii
Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, has been m...
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MDPI AG
2017-08-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/22/8/1387 |
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| author | Beibei Xiang Xiaoxue Li Yan Wang Xiaoxuan Tian Zhen Yang Lin Ma Xia Liu Yong Wang |
| author_facet | Beibei Xiang Xiaoxue Li Yan Wang Xiaoxuan Tian Zhen Yang Lin Ma Xia Liu Yong Wang |
| author_sort | Beibei Xiang |
| collection | DOAJ |
| description | Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, has been mainly tested for its anti-hepatitis effects. Here, we identify two candidate genes (SmIS1 and SmIS2) that are homologues of iridoid synthase and that are components of the secoiridoid pathway in S. mussotii. Using sequencing and phylogenetic analyses, we confirm that SmIS1 and SmIS2 contain six conserved short-chain dehydrogenases/reductase (SDR) motifs and thus belong to the P5βRs group. The two purified Escherichia coli-expressed proteins reduced 8-oxogeranial to both nepetalactol and iridodials. A comparison of the kinetic parameters of SmIS1 and SmIS2 recombinant proteins revealed that SmIS2 has a lower affinity than SmIS1 for 8-oxogeranial. Transcript levels of the two genes were analysed in three different tissues of S. mussotii using semi-quantitative RT-PCR and RT-qPCR. SmIS1 and SmIS2 expression levels were more abundant in leaves and stems. This investigation adds to our knowledge of P5βRs genes in the secoiridoid synthesis pathway and provides candidate genes for genetically improving S. mussotii by enhancing secondary metabolite production. |
| first_indexed | 2024-12-13T23:15:08Z |
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| id | doaj.art-95704b9e7a2c410b8732a2910cb35c85 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-13T23:15:08Z |
| publishDate | 2017-08-01 |
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| series | Molecules |
| spelling | doaj.art-95704b9e7a2c410b8732a2910cb35c852022-12-21T23:27:58ZengMDPI AGMolecules1420-30492017-08-01228138710.3390/molecules22081387molecules22081387Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia MussotiiBeibei Xiang0Xiaoxue Li1Yan Wang2Xiaoxuan Tian3Zhen Yang4Lin Ma5Xia Liu6Yong Wang7School of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Anshan road 312, Tianjin 300193, ChinaCollege of Life Science, Nankai University, Weijin road 94, 300071 Tianjin, ChinaSchool of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Anshan road 312, Tianjin 300193, ChinaTianjin State Key Laboratory of Modern Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Anshan road 312, Tianjin 300193, ChinaSchool of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Anshan road 312, Tianjin 300193, ChinaSchool of Chinese Materia Medica, Tianjin University of Traditional Chinese Medicine, Anshan road 312, Tianjin 300193, ChinaKey Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Ministry of Education, No. 29, 13th Street, TEDA 300457, Tianjin, ChinaCollege of Life Science, Nankai University, Weijin road 94, 300071 Tianjin, ChinaSwertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, has been mainly tested for its anti-hepatitis effects. Here, we identify two candidate genes (SmIS1 and SmIS2) that are homologues of iridoid synthase and that are components of the secoiridoid pathway in S. mussotii. Using sequencing and phylogenetic analyses, we confirm that SmIS1 and SmIS2 contain six conserved short-chain dehydrogenases/reductase (SDR) motifs and thus belong to the P5βRs group. The two purified Escherichia coli-expressed proteins reduced 8-oxogeranial to both nepetalactol and iridodials. A comparison of the kinetic parameters of SmIS1 and SmIS2 recombinant proteins revealed that SmIS2 has a lower affinity than SmIS1 for 8-oxogeranial. Transcript levels of the two genes were analysed in three different tissues of S. mussotii using semi-quantitative RT-PCR and RT-qPCR. SmIS1 and SmIS2 expression levels were more abundant in leaves and stems. This investigation adds to our knowledge of P5βRs genes in the secoiridoid synthesis pathway and provides candidate genes for genetically improving S. mussotii by enhancing secondary metabolite production.https://www.mdpi.com/1420-3049/22/8/1387Swertia mussotiimedicinal plantsecoiridoid biosynthesisiridoid synthaseprogesterone 5-β-reductaseheterologous expressionfunctional characterization |
| spellingShingle | Beibei Xiang Xiaoxue Li Yan Wang Xiaoxuan Tian Zhen Yang Lin Ma Xia Liu Yong Wang Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii Molecules Swertia mussotii medicinal plant secoiridoid biosynthesis iridoid synthase progesterone 5-β-reductase heterologous expression functional characterization |
| title | Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii |
| title_full | Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii |
| title_fullStr | Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii |
| title_full_unstemmed | Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii |
| title_short | Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii |
| title_sort | cloning and characterization of two iridoid synthase homologs from swertia mussotii |
| topic | Swertia mussotii medicinal plant secoiridoid biosynthesis iridoid synthase progesterone 5-β-reductase heterologous expression functional characterization |
| url | https://www.mdpi.com/1420-3049/22/8/1387 |
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