Multiple NADPH-cytochrome P450 reductases from Lycoris radiata involved in Amaryllidaceae alkaloids biosynthesis.

IF 3.9 2区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Plant Molecular Biology Pub Date : 2024-11-01 DOI:10.1007/s11103-024-01516-y
Yuqing Wu, Yifeng Zhang, Haitong Fan, Jie Gao, Siyu Shen, Jifan Jia, Rong Liu, Ping Su, Yating Hu, Wei Gao, Dan Li
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Abstract

Amaryllidaceae alkaloids (AAs), such as galanthamine and lycorine, are natural products of Lycoris radiata possessing various pharmacological activities including anti-acetylcholinesterase, anti-inflammatory, and antitumour activities. Elucidating the biosynthesis of these special AAs is crucial for understanding their production and potential modification for improved clinical application, of which cytochrome P450 enzymes catalyse the formation of key alkaloid skeletons and subsequent modification processes, with the NAPDH cytochrome P450 reductases (CPRs) serving as essential redox partners. This study identified three CPRs, LrCPR1, LrCPR2, and LrCPR3, encoding 700, 697 and 695 amino acids, respectively, which belong to Class II CPRs. The LrCPRs reduced cytochrome c and ferricyanide in an NADPH-dependent manner, and their activities all followed the typical Michaelis-Menten curve. In yeast, the co-expression of LrCPRs and CYP96T6 produced the galantamine-like alkaloid namely N-demethylnarwedine, suggesting that they support the catalytic activity of CYP96T6. Quantitative analysis of the transcriptional expression profiles showed that LrCPRs were expressed in all the examined tissues of L. radiata, and their gene expression patterns are consistent with other genes that may be involved in the biosynthetic pathway of AAs, including cinnamate 4-hydroxylase and phenylalanine ammonia-lyase. Our study firstly provides the functional characterization of LrCPRs in L. radiata, which will contribute to the discovery of biosynthetic pathways and heterologous production of AAs.

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枸杞中多种 NADPH-细胞色素 P450 还原酶参与金盏花科生物碱的生物合成。
Amaryllidaceae生物碱(AAs),如加兰他敏和番荔枝碱,是Lycoris radiata的天然产物,具有多种药理活性,包括抗乙酰胆碱酯酶、抗炎和抗肿瘤活性。其中细胞色素 P450 酶催化了关键生物碱骨架的形成和随后的修饰过程,而 NAPDH 细胞色素 P450 还原酶(CPRs)则是必不可少的氧化还原伙伴。这项研究发现了三种细胞色素还原酶,分别是 LrCPR1、LrCPR2 和 LrCPR3,它们分别编码 700、697 和 695 个氨基酸,属于第二类细胞色素还原酶。LrCPRs 以 NADPH 依赖性方式还原细胞色素 c 和铁氰化物,其活性均遵循典型的 Michaelis-Menten 曲线。在酵母中,LrCPRs 和 CYP96T6 的共同表达产生了类似加兰他敏的生物碱,即 N-去甲基那韦丁,这表明它们支持 CYP96T6 的催化活性。对转录表达谱的定量分析表明,LrCPRs 在辐射杉所有受检组织中均有表达,其基因表达模式与可能参与 AAs 生物合成途径的其他基因(包括肉桂酸 4-羟化酶和苯丙氨酸氨解酶)一致。我们的研究首次提供了LrCPRs的功能特征,这将有助于发现AAs的生物合成途径和异源生产。
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来源期刊
Plant Molecular Biology
Plant Molecular Biology 生物-生化与分子生物学
自引率
2.00%
发文量
95
审稿时长
1.4 months
期刊介绍: Plant Molecular Biology is an international journal dedicated to rapid publication of original research articles in all areas of plant biology.The Editorial Board welcomes full-length manuscripts that address important biological problems of broad interest, including research in comparative genomics, functional genomics, proteomics, bioinformatics, computational biology, biochemical and regulatory networks, and biotechnology. Because space in the journal is limited, however, preference is given to publication of results that provide significant new insights into biological problems and that advance the understanding of structure, function, mechanisms, or regulation. Authors must ensure that results are of high quality and that manuscripts are written for a broad plant science audience.
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