Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis

Pterocarpan glucosides are biologically active natural products, in which the glucosyl group plays an important role. However, specific pterocarpan glycosyltransferases have rarely been reported. In this study, two highly efficient glycosyltransferases, AmGT28 and AmGT44, from Astragalus membranaceu...

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Main Authors: Bai-Han Gao, Meng Zhang, Kuan Chen, Lin-Lin Wang, Ming-Ju Yao, Hong-Ye Li, Ming Xiong, Yu-Xi Huang, Yang Zhang, Quan-Li Liu, Juan Guo, Min Ye, Xue Qiao
Format: Article
Language:English
Published: Maximum Academic Press 2024-01-01
Series:Medicinal Plant Biology
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Online Access:https://www.maxapress.com/article/doi/10.48130/mpb-0024-0013
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author Bai-Han Gao
Meng Zhang
Kuan Chen
Lin-Lin Wang
Ming-Ju Yao
Hong-Ye Li
Ming Xiong
Yu-Xi Huang
Yang Zhang
Quan-Li Liu
Juan Guo
Min Ye
Xue Qiao
author_facet Bai-Han Gao
Meng Zhang
Kuan Chen
Lin-Lin Wang
Ming-Ju Yao
Hong-Ye Li
Ming Xiong
Yu-Xi Huang
Yang Zhang
Quan-Li Liu
Juan Guo
Min Ye
Xue Qiao
author_sort Bai-Han Gao
collection DOAJ
description Pterocarpan glucosides are biologically active natural products, in which the glucosyl group plays an important role. However, specific pterocarpan glycosyltransferases have rarely been reported. In this study, two highly efficient glycosyltransferases, AmGT28 and AmGT44, from Astragalus membranaceus, catalyzing the conversion of medicarpin (1) to medicarpin 3-O-glucoside (1a) were identified. They could also use other pterocarpans and isoflavonoids as substrates, showing a preference for pterocarpans over isoflavonoids. A new isoflavone 7-O-glucoside 5a was prepared through a scaled-up enzymatic reaction. Furthermore, AmGT44 was expressed in E. coli to establish a whole-cell biocatalytic system (500 μL with 0.1 mM substrate), which could glucosylate pterocarpans 1-3 with a conversion rate of up to 100% and a titer of 30-38 μg/mL within 25 h. The titer of maackiain 3-O-glucoside (2a) could reach 78.66 μg/mL in a 10-mL system. The study provides efficient catalytic tools for the biosynthesis of pterocarpan glycosides.
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spelling doaj-art-6c1b69901f7c484db2bb80e91132dbb62025-08-20T03:18:23ZengMaximum Academic PressMedicinal Plant Biology2835-69692024-01-01311610.48130/mpb-0024-0013mpb-0024-0013Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysisBai-Han Gao0Meng Zhang1Kuan Chen2Lin-Lin Wang3Ming-Ju Yao4Hong-Ye Li5Ming Xiong6Yu-Xi Huang7Yang Zhang8Quan-Li Liu9Juan Guo10Min Ye11Xue Qiao12State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaQingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences. No. 189 Songling Road, Laoshan District, Qingdao 266101, ChinaState Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, National Resource Center for Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100000, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaState Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, 38 Xueyuan Road, Beijing 100191, ChinaPterocarpan glucosides are biologically active natural products, in which the glucosyl group plays an important role. However, specific pterocarpan glycosyltransferases have rarely been reported. In this study, two highly efficient glycosyltransferases, AmGT28 and AmGT44, from Astragalus membranaceus, catalyzing the conversion of medicarpin (1) to medicarpin 3-O-glucoside (1a) were identified. They could also use other pterocarpans and isoflavonoids as substrates, showing a preference for pterocarpans over isoflavonoids. A new isoflavone 7-O-glucoside 5a was prepared through a scaled-up enzymatic reaction. Furthermore, AmGT44 was expressed in E. coli to establish a whole-cell biocatalytic system (500 μL with 0.1 mM substrate), which could glucosylate pterocarpans 1-3 with a conversion rate of up to 100% and a titer of 30-38 μg/mL within 25 h. The titer of maackiain 3-O-glucoside (2a) could reach 78.66 μg/mL in a 10-mL system. The study provides efficient catalytic tools for the biosynthesis of pterocarpan glycosides.https://www.maxapress.com/article/doi/10.48130/mpb-0024-0013astragalus membranaceuswhole-cell biocatalysispterocarpan glycosyltransferases
spellingShingle Bai-Han Gao
Meng Zhang
Kuan Chen
Lin-Lin Wang
Ming-Ju Yao
Hong-Ye Li
Ming Xiong
Yu-Xi Huang
Yang Zhang
Quan-Li Liu
Juan Guo
Min Ye
Xue Qiao
Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
Medicinal Plant Biology
astragalus membranaceus
whole-cell biocatalysis
pterocarpan glycosyltransferases
title Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
title_full Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
title_fullStr Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
title_full_unstemmed Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
title_short Characterization of two pterocarpan glycosyltransferases in Astragalus membranaceus and their application in whole-cell biocatalysis
title_sort characterization of two pterocarpan glycosyltransferases in astragalus membranaceus and their application in whole cell biocatalysis
topic astragalus membranaceus
whole-cell biocatalysis
pterocarpan glycosyltransferases
url https://www.maxapress.com/article/doi/10.48130/mpb-0024-0013
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