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Archive > Volume 17 Issue 1 > 2025,17(1):178-188. DOI:https://doi.org/10.1016/j.chmed.2024.11.003 Prev Next

代謝組和轉(zhuǎn)錄組關(guān)聯(lián)研究揭示了黃檗中特異的芐基異喹啉生物堿的生物合成過程

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Metabolome and transcriptome association study reveals biosynthesis of specialized benzylisoquinoline alkaloids in Phellodendron amurense

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    摘要:
    目的：芐基異喹啉生物堿（BIAs）具有藥理功能和臨床用途。BIAs主要分布于無患子科的毛茛目和黃檗屬的植物物種中。人們對毛茛目植物中BIAs的生物合成進(jìn)行了深入研究，但對黃檗屬植物中BIAs的積累機(jī)制卻知之甚少。本研究旨在揭示黃檗中BIAs的生物合成途徑。 方法：在此，我們對黃檗18個組織的轉(zhuǎn)錄組和代謝組數(shù)據(jù)進(jìn)行了測序和分析。利用加權(quán)基因共表達(dá)網(wǎng)絡(luò)分析（WGCNA）確定了參與BIAs生物合成的候選基因。在大腸桿菌中表達(dá)含有候選基因的重組質(zhì)粒，以評估酶活性。 結(jié)果：同源基因在同義位點(diǎn)的同義替換表明至少發(fā)生過一次全基因組復(fù)制事件。本研究提出了黃檗中潛在的BIAs的生物合成途徑，并篩選出了與BIA生物合成相關(guān)的潛在候選基因，包括2個PR10/Bet v1成員、14個CYP450和33個甲基轉(zhuǎn)移酶。其中一個PR10/Bet v1成員被鑒定為去甲烏藥堿合酶，它可以催化多巴胺和4-羥基苯乙醛生成（S）-去甲烏藥堿。 結(jié)論：我們的研究為非毛茛目物種中BIAs的生物合成和進(jìn)化提供了重要的見解。

    Abstract:
    Objective: Benzylisoquinoline alkaloids (BIAs) have pharmacological functions and clinical use. BIAs are mainly distributed in plant species across the order Ranunculales and the genus Phellodendron from Sapindales. The BIA biosynthesis has been intensively investigated in Ranunculales species. However, the accumulation mechanism of BIAs in Phellodendron is largely unknown. The aim of this study is to unravel the biosynthetic pathways of BIAs in Phellodendron amurens. Methods: The transcriptome and metabolome data from 18 different tissues of P. amurense were meticulously sequenced and subsequently subjected to a thorough analysis. Weighted gene co-expression network analysis (WGCNA), a powerful systems biology approach that facilitates the construction and subsequent analysis of co-expression networks, was utilized to identify candidate genes involved in BIAs biosynthesis. Following this, recombinant plasmids containing candidate genes were expressed in Escherichia coli, a widely used prokaryotic expression system. The purpose of this genetic engineering endeavor was to express the candidate genes within the bacteria, thereby enabling the assessment of the resultant enzyme activity. Results: The synonymous substitutions per synonymous site for paralogs indicated that at least one whole genome duplication event has occurred. The potential BIA biosynthetic pathway of P. amurense was proposed, and two PR10/Bet v1 members, 14 CYP450s, and 33 methyltransferases were selected as related to BIA biosynthesis. One PR10/Bet v1 was identified as norcoclaurine synthase, which could catalyze dopamine and 4-hydroxyphenylacetaldehyde into (S)-norcoclaurine. Conclusion: Our studies provide important insights into the biosynthesis and evolution of BIAs in non-Ranunculales species.

    關(guān)鍵詞:
    芐基異喹啉生物堿;生物合成;進(jìn)化;去甲烏藥堿合成酶;黃檗

    Keywords:


    Project Supported:
    This research was funded by grants from the National Key Research and Development Project ‘‘research and demonstration of collection, screening, and breeding technology of ginseng and other genuine medicinal materials” (No. 2021YFD1600901), National Natural Science Foundation of China (No. 82274037), and Heilongjiang Touyan Innovation Team Program (No. HLJTYTP2019001).

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