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Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd

Panax notoginseng is one of the most valuable traditional Chinese herbs. The main active ingredients, dammarane-type ginsenosides, show multiple pharmacological activities. Recently, the key UDP-dependent glycosyltransferases (UGTs) involved in the biosynthesis of common ginsenosides have been widel...

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Autores principales: Lin, Yuan, Wang, Yi Na, Zhang, Guang Hui, Chen, Geng, Yang, Qing Hui, Hao, Bing, Yang, Sheng Chao
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315489/
https://www.ncbi.nlm.nih.gov/pubmed/37405161
http://dx.doi.org/10.3389/fmicb.2023.1191102
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author Lin, Yuan
Wang, Yi Na
Zhang, Guang Hui
Chen, Geng
Yang, Qing Hui
Hao, Bing
Yang, Sheng Chao
author_facet Lin, Yuan
Wang, Yi Na
Zhang, Guang Hui
Chen, Geng
Yang, Qing Hui
Hao, Bing
Yang, Sheng Chao
author_sort Lin, Yuan
collection PubMed
description Panax notoginseng is one of the most valuable traditional Chinese herbs. The main active ingredients, dammarane-type ginsenosides, show multiple pharmacological activities. Recently, the key UDP-dependent glycosyltransferases (UGTs) involved in the biosynthesis of common ginsenosides have been widely studied. However, only a few UGTs that catalyze ginsenoside formation have been reported. This study further investigated the new catalytic function of 10 characterized UGTs from the public database. PnUGT31(PnUGT94B2) and PnUGT53 (PnUGT71B8)exhibited promiscuous sugar-donor specificity of UDP-glucose and UDP-xylose, which could catalyze the glycosylation of C20-OH sites and elongation of the sugar chain at the C3 and/or C20 sites. We further analyzed the expression patterns in P. notoginseng and predicted the catalytic mechanisms of PnUGT31 and PnUGT53 using molecular docking simulations. Moreover, different gene modules were built to increase the yield of ginsenosides in engineered yeast. The metabolic flow of the proginsenediol (PPD) synthetic pathway was enhanced by LPPDS gene modules based on the engineered strain. The resulting yeast was constructed to produce 1.72 g/L PPD in a shaking flask, but cell growth was significantly inhibited. EGH and LKG gene modules were constructed to achieve high-level production of dammarane-type ginsenosides. The production of G-Rg3 controlled by LKG modules increased 3.84 times (254.07 mg/ L), whereas the G-Rd titer reached 56.68 mg/L after 96 h in shaking flask culture under the control of all modules, both of which yielded the highest values for known microbes.
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spelling pubmed-103154892023-07-04 Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd Lin, Yuan Wang, Yi Na Zhang, Guang Hui Chen, Geng Yang, Qing Hui Hao, Bing Yang, Sheng Chao Front Microbiol Microbiology Panax notoginseng is one of the most valuable traditional Chinese herbs. The main active ingredients, dammarane-type ginsenosides, show multiple pharmacological activities. Recently, the key UDP-dependent glycosyltransferases (UGTs) involved in the biosynthesis of common ginsenosides have been widely studied. However, only a few UGTs that catalyze ginsenoside formation have been reported. This study further investigated the new catalytic function of 10 characterized UGTs from the public database. PnUGT31(PnUGT94B2) and PnUGT53 (PnUGT71B8)exhibited promiscuous sugar-donor specificity of UDP-glucose and UDP-xylose, which could catalyze the glycosylation of C20-OH sites and elongation of the sugar chain at the C3 and/or C20 sites. We further analyzed the expression patterns in P. notoginseng and predicted the catalytic mechanisms of PnUGT31 and PnUGT53 using molecular docking simulations. Moreover, different gene modules were built to increase the yield of ginsenosides in engineered yeast. The metabolic flow of the proginsenediol (PPD) synthetic pathway was enhanced by LPPDS gene modules based on the engineered strain. The resulting yeast was constructed to produce 1.72 g/L PPD in a shaking flask, but cell growth was significantly inhibited. EGH and LKG gene modules were constructed to achieve high-level production of dammarane-type ginsenosides. The production of G-Rg3 controlled by LKG modules increased 3.84 times (254.07 mg/ L), whereas the G-Rd titer reached 56.68 mg/L after 96 h in shaking flask culture under the control of all modules, both of which yielded the highest values for known microbes. Frontiers Media S.A. 2023-06-19 /pmc/articles/PMC10315489/ /pubmed/37405161 http://dx.doi.org/10.3389/fmicb.2023.1191102 Text en Copyright © 2023 Lin, Wang, Zhang, Chen, Yang, Hao and Yang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Lin, Yuan
Wang, Yi Na
Zhang, Guang Hui
Chen, Geng
Yang, Qing Hui
Hao, Bing
Yang, Sheng Chao
Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title_full Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title_fullStr Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title_full_unstemmed Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title_short Reconstruction of engineered yeast factory for high yield production of ginsenosides Rg3 and Rd
title_sort reconstruction of engineered yeast factory for high yield production of ginsenosides rg3 and rd
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10315489/
https://www.ncbi.nlm.nih.gov/pubmed/37405161
http://dx.doi.org/10.3389/fmicb.2023.1191102
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