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Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus

Acetylation contributes to the bioactivity of numerous medicinally important natural products. However, little is known about the acetylation on sugar moieties. Here we report a saponin acetyltransferase from Astragalus membranaceus. AmAT7-3 is discovered through a stepwise gene mining approach and...

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Autores principales: Wang, Linlin, Jiang, Zhihui, Zhang, Jiahe, Chen, Kuan, Zhang, Meng, Wang, Zilong, Wang, Binju, Ye, Min, Qiao, Xue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10519980/
https://www.ncbi.nlm.nih.gov/pubmed/37749089
http://dx.doi.org/10.1038/s41467-023-41599-7
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author Wang, Linlin
Jiang, Zhihui
Zhang, Jiahe
Chen, Kuan
Zhang, Meng
Wang, Zilong
Wang, Binju
Ye, Min
Qiao, Xue
author_facet Wang, Linlin
Jiang, Zhihui
Zhang, Jiahe
Chen, Kuan
Zhang, Meng
Wang, Zilong
Wang, Binju
Ye, Min
Qiao, Xue
author_sort Wang, Linlin
collection PubMed
description Acetylation contributes to the bioactivity of numerous medicinally important natural products. However, little is known about the acetylation on sugar moieties. Here we report a saponin acetyltransferase from Astragalus membranaceus. AmAT7-3 is discovered through a stepwise gene mining approach and characterized as the xylose C3′/C4′-O-acetyltransferse of astragaloside IV (1). To elucidate its catalytic mechanism, complex crystal structures of AmAT7-3/1 and AmAT7-3(A310G)/1 are obtained, which reveal a large active pocket decided by a specific sequence AADAG. Combining with QM/MM computation, the regiospecificity of AmAT7-3 is determined by sugar positioning modulated by surrounding amino acids including #A310 and #L290. Furthermore, a small mutant library is built using semi-rational design, where variants A310G and A310W are found to catalyze specific C3′-O and C4′-O acetylation, respectively. AmAT7-3 and its variants are also employed to acetylate other bioactive saponins. This work expands the understanding of saponin acetyltransferases, and provide efficient catalytic tools for saponin acetylation.
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spelling pubmed-105199802023-09-27 Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus Wang, Linlin Jiang, Zhihui Zhang, Jiahe Chen, Kuan Zhang, Meng Wang, Zilong Wang, Binju Ye, Min Qiao, Xue Nat Commun Article Acetylation contributes to the bioactivity of numerous medicinally important natural products. However, little is known about the acetylation on sugar moieties. Here we report a saponin acetyltransferase from Astragalus membranaceus. AmAT7-3 is discovered through a stepwise gene mining approach and characterized as the xylose C3′/C4′-O-acetyltransferse of astragaloside IV (1). To elucidate its catalytic mechanism, complex crystal structures of AmAT7-3/1 and AmAT7-3(A310G)/1 are obtained, which reveal a large active pocket decided by a specific sequence AADAG. Combining with QM/MM computation, the regiospecificity of AmAT7-3 is determined by sugar positioning modulated by surrounding amino acids including #A310 and #L290. Furthermore, a small mutant library is built using semi-rational design, where variants A310G and A310W are found to catalyze specific C3′-O and C4′-O acetylation, respectively. AmAT7-3 and its variants are also employed to acetylate other bioactive saponins. This work expands the understanding of saponin acetyltransferases, and provide efficient catalytic tools for saponin acetylation. Nature Publishing Group UK 2023-09-25 /pmc/articles/PMC10519980/ /pubmed/37749089 http://dx.doi.org/10.1038/s41467-023-41599-7 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This 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/) .
spellingShingle Article
Wang, Linlin
Jiang, Zhihui
Zhang, Jiahe
Chen, Kuan
Zhang, Meng
Wang, Zilong
Wang, Binju
Ye, Min
Qiao, Xue
Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title_full Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title_fullStr Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title_full_unstemmed Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title_short Characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from Astragalus membranaceus
title_sort characterization and structure-based protein engineering of a regiospecific saponin acetyltransferase from astragalus membranaceus
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10519980/
https://www.ncbi.nlm.nih.gov/pubmed/37749089
http://dx.doi.org/10.1038/s41467-023-41599-7
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