Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>

Ginseng (<i>Panax ginseng</i> C.A. Mey.) is a precious Chinese traditional medicine, for which ginsenosides are the most important medicinal ingredients. Cytochrome P450 enzymes (CYP450) and their primary redox molecular companion NADPH cytochrome P450 reductase (CPR) play a key role in...

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Autores principales:	Xian Zou, Yue Zhang, Xu Zeng, Tuo Liu, Gui Li, Yuxin Dai, Yuanzhu Xie, Zhiyong Luo
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	NADPH-cytochrome P450 reductase ginsenoside methyl jasmonate prokaryotic expression PgCPR1 and PgCPR2 Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/1c51639e42ab4fe6ac51cf7d52f9124c
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spelling	oai:doaj.org-article:1c51639e42ab4fe6ac51cf7d52f9124c2021-11-11T18:37:18ZMolecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>10.3390/molecules262166541420-3049https://doaj.org/article/1c51639e42ab4fe6ac51cf7d52f9124c2021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/21/6654https://doaj.org/toc/1420-3049Ginseng (<i>Panax ginseng</i> C.A. Mey.) is a precious Chinese traditional medicine, for which ginsenosides are the most important medicinal ingredients. Cytochrome P450 enzymes (CYP450) and their primary redox molecular companion NADPH cytochrome P450 reductase (CPR) play a key role in ginsenoside biosynthesis pathway. However, systematic studies of CPR genes in ginseng have not been reported. Numerous studies on ginsenoside synthesis biology still use <i>Arabidopsis</i> CPR (AtCPR1) as a reductase. In this study, we isolated two CPR genes (<i>PgCPR1</i>, <i>PgCPR2</i>) from ginseng adventitious roots. Phylogenetic tree analysis showed that both PgCPR1 and PgCPR2 are grouped in classⅡ of dicotyledonous CPR. Enzyme experiments showed that recombinant proteins PgCPR1, PgCPR2 and AtCPR1 can reduce cytochrome c and ferricyanide with NADPH as the electron donor, and PgCPR1 had the highest enzymatic activities. Quantitative real-time PCR analysis showed that PgCPR1 and PgCPR2 transcripts were detected in all examined tissues of <i>Panax ginseng</i> and both showed higher expression in stem and main root. Expression levels of the PgCPR1 and PgCPR2s were both induced after a methyl jasmonate (MeJA) treatment and its pattern matched with ginsenoside accumulation. The present investigation suggested PgCPR1 and PgCPR2 are associated with the biosynthesis of ginsenoside. This report will assist in future CPR family studies and ultimately improving ginsenoside production through transgenic engineering and synthetic biology.Xian ZouYue ZhangXu ZengTuo LiuGui LiYuxin DaiYuanzhu XieZhiyong LuoMDPI AGarticleNADPH-cytochrome P450 reductaseginsenosidemethyl jasmonateprokaryotic expressionPgCPR1 and PgCPR2Organic chemistryQD241-441ENMolecules, Vol 26, Iss 6654, p 6654 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	NADPH-cytochrome P450 reductase ginsenoside methyl jasmonate prokaryotic expression PgCPR1 and PgCPR2 Organic chemistry QD241-441
spellingShingle	NADPH-cytochrome P450 reductase ginsenoside methyl jasmonate prokaryotic expression PgCPR1 and PgCPR2 Organic chemistry QD241-441 Xian Zou Yue Zhang Xu Zeng Tuo Liu Gui Li Yuxin Dai Yuanzhu Xie Zhiyong Luo Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
description	Ginseng (<i>Panax ginseng</i> C.A. Mey.) is a precious Chinese traditional medicine, for which ginsenosides are the most important medicinal ingredients. Cytochrome P450 enzymes (CYP450) and their primary redox molecular companion NADPH cytochrome P450 reductase (CPR) play a key role in ginsenoside biosynthesis pathway. However, systematic studies of CPR genes in ginseng have not been reported. Numerous studies on ginsenoside synthesis biology still use <i>Arabidopsis</i> CPR (AtCPR1) as a reductase. In this study, we isolated two CPR genes (<i>PgCPR1</i>, <i>PgCPR2</i>) from ginseng adventitious roots. Phylogenetic tree analysis showed that both PgCPR1 and PgCPR2 are grouped in classⅡ of dicotyledonous CPR. Enzyme experiments showed that recombinant proteins PgCPR1, PgCPR2 and AtCPR1 can reduce cytochrome c and ferricyanide with NADPH as the electron donor, and PgCPR1 had the highest enzymatic activities. Quantitative real-time PCR analysis showed that PgCPR1 and PgCPR2 transcripts were detected in all examined tissues of <i>Panax ginseng</i> and both showed higher expression in stem and main root. Expression levels of the PgCPR1 and PgCPR2s were both induced after a methyl jasmonate (MeJA) treatment and its pattern matched with ginsenoside accumulation. The present investigation suggested PgCPR1 and PgCPR2 are associated with the biosynthesis of ginsenoside. This report will assist in future CPR family studies and ultimately improving ginsenoside production through transgenic engineering and synthetic biology.
format	article
author	Xian Zou Yue Zhang Xu Zeng Tuo Liu Gui Li Yuxin Dai Yuanzhu Xie Zhiyong Luo
author_facet	Xian Zou Yue Zhang Xu Zeng Tuo Liu Gui Li Yuxin Dai Yuanzhu Xie Zhiyong Luo
author_sort	Xian Zou
title	Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
title_short	Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
title_full	Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
title_fullStr	Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
title_full_unstemmed	Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>
title_sort	molecular cloning and identification of nadph cytochrome p450 reductase from <i>panax ginseng</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/1c51639e42ab4fe6ac51cf7d52f9124c
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Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from <i>Panax ginseng</i>

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