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马蹄金中 4-香豆酸辅酶 A 连接酶的分子克隆和功能分析及其在木质素和黄烷酮生物合成中的作用。

Molecular cloning and functional analysis of 4-coumarate: CoA ligases from Marchantia paleacea and their roles in lignin and flavanone biosynthesis.

机构信息

Key Laboratory of Chemical Biology of Natural Products, Ministry of Education, School of Pharmaceutical Sciences, Shandong University, Jinan, Shandong, China.

Shandong Provincial Clinical Research Center for Emergency and Critical Care Medicine, Jinan, Shan-dong, China.

出版信息

PLoS One. 2024 Jan 8;19(1):e0296079. doi: 10.1371/journal.pone.0296079. eCollection 2024.

DOI:10.1371/journal.pone.0296079
PMID:38190396
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10773943/
Abstract

Phenylpropanoids play important roles in plant physiology and the enzyme 4-coumarate: coenzyme A ligase (4CL) catalyzes the formation of thioesters. Despite extensive characterization in various plants, the functions of 4CLs in the liverwort Marchantia paleacea remain unknown. Here, four 4CLs from M. paleacea were isolated and functionally analyzed. Heterologous expression in Escherichia coli indicated the presence of different enzymatic activities in the four enzymes. Mp4CL1 and Mp4CL2 were able to convert caffeic, p-coumaric, cinnamic, ferulic, dihydro-p-coumaric, and 5-hydroxyferulic acids to their corresponding CoA esters, while Mp4CL3 and Mp4CL4 catalyzed none. Mp4CL1 transcription was induced when M. paleacea thalli were treated with methyl jasmonate (MeJA). The overexpression of Mp4CL1 increased the levels of lignin in transgenic Arabidopsis. In addition, we reconstructed the flavanone biosynthetic pathway in E. coli. The pathway comprised Mp4CL1, co-expressed with chalcone synthase (CHS) from different plant species, and the efficiency of biosynthesis was optimal when both the 4CL and CHS were obtained from the same species M. paleacea.

摘要

苯丙烷类在植物生理学中起着重要作用，而酶 4-香豆酸：辅酶 A 连接酶（4CL）则催化硫酯的形成。尽管在各种植物中得到了广泛的研究，但在叶苔纲植物 Marchantia paleacea 中 4CL 的功能仍然未知。本文从 M. paleacea 中分离并功能分析了四个 4CL。在大肠杆菌中的异源表达表明，这四种酶具有不同的酶活性。Mp4CL1 和 Mp4CL2 能够将咖啡酸、对香豆酸、肉桂酸、阿魏酸、二氢对香豆酸和 5-羟基阿魏酸转化为相应的 CoA 酯，而 Mp4CL3 和 Mp4CL4 则不能。当 M. paleacea 叶状体受到茉莉酸甲酯（MeJA）处理时，Mp4CL1 的转录被诱导。Mp4CL1 的过表达增加了转基因拟南芥中木质素的水平。此外，我们在大肠杆菌中重建了黄烷酮生物合成途径。该途径包含 Mp4CL1，与来自不同植物物种的查尔酮合酶（CHS）共表达，当 4CL 和 CHS 均来自同一物种 M. paleacea 时，生物合成的效率最佳。

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