Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from Panax ginseng.
Molecular Cloning and Identification of NADPH Cytochrome P450 Reductase from Panax ginseng.
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人参NADPH细胞色素P450还原酶的分子克隆及鉴定
DOI：
10.3390/molecules26216654
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发表时间：
2021-11-03
期刊：
影响因子：
--
通讯作者：
Luo Z
中科院分区：
文献类型：
--
作者：
Zou X;Zhang Y;Zeng X;Liu T;Li G;Dai Y;Xie Y;Luo Z
人参（Panax Ginseng C.A. Mey.）是一种珍贵的中药，人参皂苷是其最重要的药用成分。细胞色素P450酶（CYP450）及其主要氧化还原分子伴侣NADPH细胞色素P450还原酶（CPR）在人参皂苷生物合成途径中起关键作用。然而，对人参中CPR基因的系统研究尚未见报道。许多人参皂苷合成生物学研究仍然使用拟南芥CPR （AtCPR1）作为还原酶。本研究从人参不定根中分离出两个CPR基因PgCPR1、PgCPR2。系统进化树分析显示，PgCPR1和PgCPR2均归属于双子叶植物CPRⅡ纲。酶促实验表明，重组蛋白PgCPR1、PgCPR2和AtCPR1均能以NADPH为电子供体还原细胞色素c和铁氰化物，其中PgCPR1酶促活性最高。实时荧光定量PCR分析显示，PgCPR1和PgCPR2转录本在人参各组织中均有表达，且在人参茎和主根中表达量均较高。在茉莉酸甲酯（MeJA）处理后，PgCPR1和PgCPR2s的表达水平均被诱导，其表达模式与人参皂苷积累相匹配。目前的研究表明PgCPR1和PgCPR2与人参皂苷的生物合成有关。该报告将有助于未来的CPR家族研究，并最终通过转基因工程和合成生物学提高人参皂苷的产量。
Ginseng (Panax ginseng 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 Arabidopsis CPR (AtCPR1) as a reductase. In this study, we isolated two CPR genes (PgCPR1, PgCPR2) 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 Panax ginseng 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.
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DOI：
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发表时间：
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期刊：
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