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Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli

BACKGROUND: Flavonoid C-glycosides have many beneficial effects and are widely used in food and medicine. However, plants contain a limited number of flavonoid C-glycosides, and it is challenging to create these substances chemically. RESULTS: To screen more robust C-glycosyltransferases (CGTs) for...

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Autores principales: Ni, Rong, Liu, Xin-Yan, Zhang, Jiao-Zhen, Fu, Jie, Tan, Hui, Zhu, Ting-Ting, Zhang, Jing, Wang, Hai-Long, Lou, Hong-Xiang, Cheng, Ai-Xia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9563126/
https://www.ncbi.nlm.nih.gov/pubmed/36242071
http://dx.doi.org/10.1186/s12934-022-01940-z
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author Ni, Rong
Liu, Xin-Yan
Zhang, Jiao-Zhen
Fu, Jie
Tan, Hui
Zhu, Ting-Ting
Zhang, Jing
Wang, Hai-Long
Lou, Hong-Xiang
Cheng, Ai-Xia
author_facet Ni, Rong
Liu, Xin-Yan
Zhang, Jiao-Zhen
Fu, Jie
Tan, Hui
Zhu, Ting-Ting
Zhang, Jing
Wang, Hai-Long
Lou, Hong-Xiang
Cheng, Ai-Xia
author_sort Ni, Rong
collection PubMed
description BACKGROUND: Flavonoid C-glycosides have many beneficial effects and are widely used in food and medicine. However, plants contain a limited number of flavonoid C-glycosides, and it is challenging to create these substances chemically. RESULTS: To screen more robust C-glycosyltransferases (CGTs) for the biosynthesis of flavonoid C-glycosides, one CGT enzyme from Stenoloma chusanum (ScCGT1) was characterized. Biochemical analyses revealed that ScCGT1 showed the C-glycosylation activity for phloretin, 2-hydroxynaringenin, and 2-hydroxyeriodictyol. Structure modeling and mutagenesis experiments indicated that the glycosylation of ScCGT1 may be initiated by the synergistic action of conserved residue His26 and Asp14. The P164T mutation increased C-glycosylation activity by forming a hydrogen bond with the sugar donor. Furthermore, when using phloretin as a substrate, the extracellular nothofagin production obtained from the Escherichia coli strain ScCGT1-P164T reached 38 mg/L, which was 2.3-fold higher than that of the wild-type strain. Finally, it is proved that the coupling catalysis of CjFNS I/F2H and ScCGT1-P164T could convert naringenin into vitexin and isovitexin. CONCLUSION: This is the first time that C-glycosyltransferase has been characterized from fern species and provides a candidate gene and strategy for the efficient production of bioactive C-glycosides using enzyme catalysis and metabolic engineering. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01940-z.
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spelling pubmed-95631262022-10-15 Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli Ni, Rong Liu, Xin-Yan Zhang, Jiao-Zhen Fu, Jie Tan, Hui Zhu, Ting-Ting Zhang, Jing Wang, Hai-Long Lou, Hong-Xiang Cheng, Ai-Xia Microb Cell Fact Research BACKGROUND: Flavonoid C-glycosides have many beneficial effects and are widely used in food and medicine. However, plants contain a limited number of flavonoid C-glycosides, and it is challenging to create these substances chemically. RESULTS: To screen more robust C-glycosyltransferases (CGTs) for the biosynthesis of flavonoid C-glycosides, one CGT enzyme from Stenoloma chusanum (ScCGT1) was characterized. Biochemical analyses revealed that ScCGT1 showed the C-glycosylation activity for phloretin, 2-hydroxynaringenin, and 2-hydroxyeriodictyol. Structure modeling and mutagenesis experiments indicated that the glycosylation of ScCGT1 may be initiated by the synergistic action of conserved residue His26 and Asp14. The P164T mutation increased C-glycosylation activity by forming a hydrogen bond with the sugar donor. Furthermore, when using phloretin as a substrate, the extracellular nothofagin production obtained from the Escherichia coli strain ScCGT1-P164T reached 38 mg/L, which was 2.3-fold higher than that of the wild-type strain. Finally, it is proved that the coupling catalysis of CjFNS I/F2H and ScCGT1-P164T could convert naringenin into vitexin and isovitexin. CONCLUSION: This is the first time that C-glycosyltransferase has been characterized from fern species and provides a candidate gene and strategy for the efficient production of bioactive C-glycosides using enzyme catalysis and metabolic engineering. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-022-01940-z. BioMed Central 2022-10-14 /pmc/articles/PMC9563126/ /pubmed/36242071 http://dx.doi.org/10.1186/s12934-022-01940-z Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Ni, Rong
Liu, Xin-Yan
Zhang, Jiao-Zhen
Fu, Jie
Tan, Hui
Zhu, Ting-Ting
Zhang, Jing
Wang, Hai-Long
Lou, Hong-Xiang
Cheng, Ai-Xia
Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title_full Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title_fullStr Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title_full_unstemmed Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title_short Identification of a flavonoid C-glycosyltransferase from fern species Stenoloma chusanum and the application in synthesizing flavonoid C-glycosides in Escherichia coli
title_sort identification of a flavonoid c-glycosyltransferase from fern species stenoloma chusanum and the application in synthesizing flavonoid c-glycosides in escherichia coli
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9563126/
https://www.ncbi.nlm.nih.gov/pubmed/36242071
http://dx.doi.org/10.1186/s12934-022-01940-z
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