Cargando…
Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids
Isatis indigotica accumulates several active substances, including C-glycosylflavonoids, which have important pharmacological activities and health benefits. However, enzymes catalyzing the methylation step of C-glycosylflavonoids in I. indigotica remain unknown. In this study, three O-methyltransfe...
Autores principales: | , , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9437721/ https://www.ncbi.nlm.nih.gov/pubmed/36072835 http://dx.doi.org/10.1093/hr/uhac140 |
_version_ | 1784781681032429568 |
---|---|
author | Tan, Yuping Yang, Jian Jiang, Yinyin Sun, Shufu Wei, Xiaoyan Wang, Ruishan Bu, Junling Li, Dayong Kang, Liping Chen, Tong Guo, Juan Cui, Guanghong Tang, Jinfu Huang, Luqi |
author_facet | Tan, Yuping Yang, Jian Jiang, Yinyin Sun, Shufu Wei, Xiaoyan Wang, Ruishan Bu, Junling Li, Dayong Kang, Liping Chen, Tong Guo, Juan Cui, Guanghong Tang, Jinfu Huang, Luqi |
author_sort | Tan, Yuping |
collection | PubMed |
description | Isatis indigotica accumulates several active substances, including C-glycosylflavonoids, which have important pharmacological activities and health benefits. However, enzymes catalyzing the methylation step of C-glycosylflavonoids in I. indigotica remain unknown. In this study, three O-methyltransferases (OMTs) were identified from I. indigotica that have the capacity for O-methylation of the C-glycosylflavonoid isoorientin. The Type II OMTs IiOMT1 and IiOMT2 efficiently catalyze isoorientin to form isoscoparin, and decorate one of the aromatic vicinal hydroxyl groups on flavones and methylate the C6, C8, and 3′-hydroxyl positions to form oroxylin A, wogonin, and chrysoeriol, respectively. However, the Type I OMT IiOMT3 exhibited broader substrate promiscuity and methylated the C7 and 3′-hydroxyl positions of flavonoids. Further site-directed mutagenesis studies demonstrated that five amino acids of IiOMT1/IiOMT2 (D121/D100, D173/D149, A174/A150R, N200/N176, and D248/D233) were critical residues for their catalytic activity. Additionally, only transient overexpression of Type II OMTs IiOMT1 and IiOMT2 in Nicotiana benthamiana significantly increased isoscoparin accumulation, indicating that the Type II OMTs IiOMT1 and IiOMT2 could catalyze the methylation step of C-glycosylflavonoid, isoorientin at the 3′-hydroxyl position. This study provides insights into the biosynthesis of methylated C-glycosylflavonoids, and IiOMTs could be promising catalysts in the synthesis of bioactive compounds. |
format | Online Article Text |
id | pubmed-9437721 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-94377212022-09-06 Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids Tan, Yuping Yang, Jian Jiang, Yinyin Sun, Shufu Wei, Xiaoyan Wang, Ruishan Bu, Junling Li, Dayong Kang, Liping Chen, Tong Guo, Juan Cui, Guanghong Tang, Jinfu Huang, Luqi Hortic Res Article Isatis indigotica accumulates several active substances, including C-glycosylflavonoids, which have important pharmacological activities and health benefits. However, enzymes catalyzing the methylation step of C-glycosylflavonoids in I. indigotica remain unknown. In this study, three O-methyltransferases (OMTs) were identified from I. indigotica that have the capacity for O-methylation of the C-glycosylflavonoid isoorientin. The Type II OMTs IiOMT1 and IiOMT2 efficiently catalyze isoorientin to form isoscoparin, and decorate one of the aromatic vicinal hydroxyl groups on flavones and methylate the C6, C8, and 3′-hydroxyl positions to form oroxylin A, wogonin, and chrysoeriol, respectively. However, the Type I OMT IiOMT3 exhibited broader substrate promiscuity and methylated the C7 and 3′-hydroxyl positions of flavonoids. Further site-directed mutagenesis studies demonstrated that five amino acids of IiOMT1/IiOMT2 (D121/D100, D173/D149, A174/A150R, N200/N176, and D248/D233) were critical residues for their catalytic activity. Additionally, only transient overexpression of Type II OMTs IiOMT1 and IiOMT2 in Nicotiana benthamiana significantly increased isoscoparin accumulation, indicating that the Type II OMTs IiOMT1 and IiOMT2 could catalyze the methylation step of C-glycosylflavonoid, isoorientin at the 3′-hydroxyl position. This study provides insights into the biosynthesis of methylated C-glycosylflavonoids, and IiOMTs could be promising catalysts in the synthesis of bioactive compounds. Oxford University Press 2022-06-23 /pmc/articles/PMC9437721/ /pubmed/36072835 http://dx.doi.org/10.1093/hr/uhac140 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Nanjing Agricultural University https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Article Tan, Yuping Yang, Jian Jiang, Yinyin Sun, Shufu Wei, Xiaoyan Wang, Ruishan Bu, Junling Li, Dayong Kang, Liping Chen, Tong Guo, Juan Cui, Guanghong Tang, Jinfu Huang, Luqi Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title | Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title_full | Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title_fullStr | Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title_full_unstemmed | Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title_short | Identification and characterization of two Isatis indigotica O-methyltransferases methylating C-glycosylflavonoids |
title_sort | identification and characterization of two isatis indigotica o-methyltransferases methylating c-glycosylflavonoids |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9437721/ https://www.ncbi.nlm.nih.gov/pubmed/36072835 http://dx.doi.org/10.1093/hr/uhac140 |
work_keys_str_mv | AT tanyuping identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT yangjian identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT jiangyinyin identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT sunshufu identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT weixiaoyan identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT wangruishan identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT bujunling identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT lidayong identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT kangliping identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT chentong identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT guojuan identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT cuiguanghong identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT tangjinfu identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids AT huangluqi identificationandcharacterizationoftwoisatisindigoticaomethyltransferasesmethylatingcglycosylflavonoids |