A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower
To explore the complete biosynthesis process of flavonoid glycosides in safflower, specifically the key glycosyltransferase that might be involved, as well as to develop an efficient biocatalyst to synthesize flavonoid glycosides, a glycosyltransferase <i>Ct</i>UGT4, with flavonoid-<i...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/28/22/7613 |
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| author | Shuyi Qi Beixuan He Haotian Wang Yaqian Duan Lunuan Wang Yue Gao Meili Guo |
| author_facet | Shuyi Qi Beixuan He Haotian Wang Yaqian Duan Lunuan Wang Yue Gao Meili Guo |
| author_sort | Shuyi Qi |
| collection | DOAJ |
| description | To explore the complete biosynthesis process of flavonoid glycosides in safflower, specifically the key glycosyltransferase that might be involved, as well as to develop an efficient biocatalyst to synthesize flavonoid glycosides, a glycosyltransferase <i>Ct</i>UGT4, with flavonoid-<i>O</i>-glycosyltransferase activity, was identified in safflower. The fusion protein of <i>Ct</i>UGT4 was heterologously expressed in <i>Escherichia coli</i>, and the target protein was purified. The recombinant protein can catalyze quercetin to form quercetin-7-<i>O</i>-glucoside, and kaempferol to form kaempferol-3-<i>O</i> in vitro, and a series of flavones, flavonols, dihydroflavones, chalcones, and chalcone glycosides were used as substrates to generate new products. <i>Ct</i>UGT4 was expressed in the tobacco transient expression system, and the enzyme activity results showed that it could catalyze kaempferol to kaempferol-3-<i>O</i>-glucoside, and quercetin to quercetin-3-<i>O</i>-glucoside. After overexpressing <i>CtUGT4</i> in safflower, the content of quercetin-3-<i>O</i>-rutinoside in the safflower florets increased significantly, and the content of quercetin-3-<i>O</i>-glucoside also tended to increase, which preliminarily confirmed the function of <i>Ct</i>UGT4 flavonoid-<i>O</i>-glycosyltransferase. This work demonstrated the flavonoid-<i>O</i>-glycosyltransferase function of safflower <i>Ct</i>UGT4 and showed differences in the affinity for different flavonoid substrates and the regioselectivity of catalytic sites in safflower, both in vivo and in vitro, providing clues for further research regarding the function of <i>UGT</i> genes, as well as new ideas for the cultivation engineering of the directional improvement of effective metabolites in safflower. |
| first_indexed | 2024-03-09T16:34:49Z |
| format | Article |
| id | doaj.art-f3a278a1729147028b312310096d0302 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T16:34:49Z |
| publishDate | 2023-11-01 |
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| record_format | Article |
| series | Molecules |
| spelling | doaj.art-f3a278a1729147028b312310096d03022023-11-24T14:58:25ZengMDPI AGMolecules1420-30492023-11-012822761310.3390/molecules28227613A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from SafflowerShuyi Qi0Beixuan He1Haotian Wang2Yaqian Duan3Lunuan Wang4Yue Gao5Meili Guo6Department of Pharmacognosy, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaDepartment of Pharmacognosy, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaDepartment of Pharmacognosy, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaChemistry Experimental Teaching Center, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaDepartment of Pharmacognosy, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaChanghai Clinical Research Unit, The First Affiliated Hospital of Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaDepartment of Pharmacognosy, College of Pharmacy, Naval Medical University (Second Military Medical University), Shanghai 200433, ChinaTo explore the complete biosynthesis process of flavonoid glycosides in safflower, specifically the key glycosyltransferase that might be involved, as well as to develop an efficient biocatalyst to synthesize flavonoid glycosides, a glycosyltransferase <i>Ct</i>UGT4, with flavonoid-<i>O</i>-glycosyltransferase activity, was identified in safflower. The fusion protein of <i>Ct</i>UGT4 was heterologously expressed in <i>Escherichia coli</i>, and the target protein was purified. The recombinant protein can catalyze quercetin to form quercetin-7-<i>O</i>-glucoside, and kaempferol to form kaempferol-3-<i>O</i> in vitro, and a series of flavones, flavonols, dihydroflavones, chalcones, and chalcone glycosides were used as substrates to generate new products. <i>Ct</i>UGT4 was expressed in the tobacco transient expression system, and the enzyme activity results showed that it could catalyze kaempferol to kaempferol-3-<i>O</i>-glucoside, and quercetin to quercetin-3-<i>O</i>-glucoside. After overexpressing <i>CtUGT4</i> in safflower, the content of quercetin-3-<i>O</i>-rutinoside in the safflower florets increased significantly, and the content of quercetin-3-<i>O</i>-glucoside also tended to increase, which preliminarily confirmed the function of <i>Ct</i>UGT4 flavonoid-<i>O</i>-glycosyltransferase. This work demonstrated the flavonoid-<i>O</i>-glycosyltransferase function of safflower <i>Ct</i>UGT4 and showed differences in the affinity for different flavonoid substrates and the regioselectivity of catalytic sites in safflower, both in vivo and in vitro, providing clues for further research regarding the function of <i>UGT</i> genes, as well as new ideas for the cultivation engineering of the directional improvement of effective metabolites in safflower.https://www.mdpi.com/1420-3049/28/22/7613safflower<i>O</i>-glycosyltransferaseflavonoidbiosynthesis |
| spellingShingle | Shuyi Qi Beixuan He Haotian Wang Yaqian Duan Lunuan Wang Yue Gao Meili Guo A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower Molecules safflower <i>O</i>-glycosyltransferase flavonoid biosynthesis |
| title | A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower |
| title_full | A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower |
| title_fullStr | A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower |
| title_full_unstemmed | A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower |
| title_short | A Muti-Substrate Flavonol <i>O</i>-glucosyltransferases from Safflower |
| title_sort | muti substrate flavonol i o i glucosyltransferases from safflower |
| topic | safflower <i>O</i>-glycosyltransferase flavonoid biosynthesis |
| url | https://www.mdpi.com/1420-3049/28/22/7613 |
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