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Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant

Scopolia lurida, a native herbal plant species in Tibet, is one of the most effective producers of tropane alkaloids. However, the tropane alkaloid biosynthesis in this plant species of interest has yet to be studied at the molecular, biochemical, and biotechnological level. Here, we report on the i...

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Autores principales: Zhao, Kaihui, Zeng, Junlan, Zhao, Tengfei, Zhang, Haoxing, Qiu, Fei, Yang, Chunxian, Zeng, Lingjiang, Liu, Xiaoqiang, Chen, Min, Lan, Xiaozhong, Liao, Zhihua
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5650612/
https://www.ncbi.nlm.nih.gov/pubmed/29085381
http://dx.doi.org/10.3389/fpls.2017.01745
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author Zhao, Kaihui
Zeng, Junlan
Zhao, Tengfei
Zhang, Haoxing
Qiu, Fei
Yang, Chunxian
Zeng, Lingjiang
Liu, Xiaoqiang
Chen, Min
Lan, Xiaozhong
Liao, Zhihua
author_facet Zhao, Kaihui
Zeng, Junlan
Zhao, Tengfei
Zhang, Haoxing
Qiu, Fei
Yang, Chunxian
Zeng, Lingjiang
Liu, Xiaoqiang
Chen, Min
Lan, Xiaozhong
Liao, Zhihua
author_sort Zhao, Kaihui
collection PubMed
description Scopolia lurida, a native herbal plant species in Tibet, is one of the most effective producers of tropane alkaloids. However, the tropane alkaloid biosynthesis in this plant species of interest has yet to be studied at the molecular, biochemical, and biotechnological level. Here, we report on the isolation and characterization of a putative short chain dehydrogenase (SDR) gene. Sequence analysis showed that SlTRI belonged to the SDR family. Phylogenetic analysis revealed that SlTRI was clustered with the tropine-forming reductases. SlTRI and the other TA-biosynthesis genes, including putrescine N-methyltransferase (SlPMT) and hyoscyamine 6β-hydroxylase (SlH6H), were preferably or exclusively expressed in the S. lurida roots. The tissue profile of SlTRI suggested that this gene might be involved in tropane alkaloid biosynthesis. By using GC-MS, SlTRI was shown to catalyze the tropinone reduction to yield tropine, the key intermediate of tropane alkaloids. With the purified recombinant SlTRI from Escherichia coli, an enzymatic assay was carried out; its result indicated that SlTRI was a tropine-forming reductase. Finally, the role of SlTRI in promoting the tropane alkaloid biosynthesis was confirmed through metabolic engineering in S. lurida. Specifically, hairy root cultures of S. lurida were established to investigate the effects of SlTRI overexpression on tropane alkaloid accumulation. In the SlTRI-overexpressing root cultures, the hyoscyamine contents were 1.7- to 2.9-fold higher than those in control while their corresponding scopolamine contents were likewise elevated. In summary, this functional identification of SlTRI has provided for a better understanding of tropane alkaloid biosynthesis. It also provides a candidate gene for enhancing tropane alkaloid biosynthesis in S. lurida via metabolic engineering.
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spelling pubmed-56506122017-10-30 Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant Zhao, Kaihui Zeng, Junlan Zhao, Tengfei Zhang, Haoxing Qiu, Fei Yang, Chunxian Zeng, Lingjiang Liu, Xiaoqiang Chen, Min Lan, Xiaozhong Liao, Zhihua Front Plant Sci Plant Science Scopolia lurida, a native herbal plant species in Tibet, is one of the most effective producers of tropane alkaloids. However, the tropane alkaloid biosynthesis in this plant species of interest has yet to be studied at the molecular, biochemical, and biotechnological level. Here, we report on the isolation and characterization of a putative short chain dehydrogenase (SDR) gene. Sequence analysis showed that SlTRI belonged to the SDR family. Phylogenetic analysis revealed that SlTRI was clustered with the tropine-forming reductases. SlTRI and the other TA-biosynthesis genes, including putrescine N-methyltransferase (SlPMT) and hyoscyamine 6β-hydroxylase (SlH6H), were preferably or exclusively expressed in the S. lurida roots. The tissue profile of SlTRI suggested that this gene might be involved in tropane alkaloid biosynthesis. By using GC-MS, SlTRI was shown to catalyze the tropinone reduction to yield tropine, the key intermediate of tropane alkaloids. With the purified recombinant SlTRI from Escherichia coli, an enzymatic assay was carried out; its result indicated that SlTRI was a tropine-forming reductase. Finally, the role of SlTRI in promoting the tropane alkaloid biosynthesis was confirmed through metabolic engineering in S. lurida. Specifically, hairy root cultures of S. lurida were established to investigate the effects of SlTRI overexpression on tropane alkaloid accumulation. In the SlTRI-overexpressing root cultures, the hyoscyamine contents were 1.7- to 2.9-fold higher than those in control while their corresponding scopolamine contents were likewise elevated. In summary, this functional identification of SlTRI has provided for a better understanding of tropane alkaloid biosynthesis. It also provides a candidate gene for enhancing tropane alkaloid biosynthesis in S. lurida via metabolic engineering. Frontiers Media S.A. 2017-10-16 /pmc/articles/PMC5650612/ /pubmed/29085381 http://dx.doi.org/10.3389/fpls.2017.01745 Text en Copyright © 2017 Zhao, Zeng, Zhao, Zhang, Qiu, Yang, Zeng, Liu, Chen, Lan and Liao. http://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) or licensor 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 Plant Science
Zhao, Kaihui
Zeng, Junlan
Zhao, Tengfei
Zhang, Haoxing
Qiu, Fei
Yang, Chunxian
Zeng, Lingjiang
Liu, Xiaoqiang
Chen, Min
Lan, Xiaozhong
Liao, Zhihua
Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title_full Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title_fullStr Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title_full_unstemmed Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title_short Enhancing Tropane Alkaloid Production Based on the Functional Identification of Tropine-Forming Reductase in Scopolia lurida, a Tibetan Medicinal Plant
title_sort enhancing tropane alkaloid production based on the functional identification of tropine-forming reductase in scopolia lurida, a tibetan medicinal plant
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5650612/
https://www.ncbi.nlm.nih.gov/pubmed/29085381
http://dx.doi.org/10.3389/fpls.2017.01745
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