Molecular cloning, functional characterization and expression of the β-amyrin synthase gene involved in saikosaponin biosynthesis in Bupleurum chinense DC.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2023-01-01 DOI:10.1007/s13562-022-00804-2

Yanping Mao, Hua Chen, Jun Zhao, Yuchan Li, Liang Feng, Yuping Yang, Yiguan Zhang, Ping Wei, Dabin Hou

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引用次数: 1

Abstract

Bupleurum chinense DC. is a commonly used plant in traditional Chinese medicine, and saikosaponins(SSs) are the main active oleanane-typetriterpene saponins in B. chinense. β-Amyrin synthase (β-AS) is an important enzyme in oleanane-type triterpenoid saponin synthesis, but its role in saikosaponin synthesis has rarely been studied. Here, the putative β-AS gene BcBAS1(Accession No.ON890382) selected according to metabolomic and transcriptomic analyses was cloned and functionally characterized by heterologous expression in Escherichia coli and Pichia pastoris, and its subcellular localization and expression patterns were examined. The molecular weight of the BcBAS1 recombinant protein was approximately 87 kDa, and this protein could catalyse the production of β-amyrin, the precursor of SSs. Furthermore, BcBAS1 was located in the cytosol, and relative expression in four tissues of the four genotypes was positively correlated with SSa and SSd contents. Our results indicate that BcBAS1 is a β-AS gene and may play an important role in saikosaponin biosynthesis and regulation. This study sheds light on the role of β-AS genes in the synthesis of SSs and provides insights for the metabolic engineering of SSs.

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柴胡皂苷合成相关β-amyrin合成酶基因的克隆、功能表征及表达

柴胡。柴草皂苷(saikosaponins, ss)是柴草中主要的齐墩烷型三萜活性皂苷。β-Amyrin合成酶(β-AS)是齐墩墩烷型三萜皂苷合成中的重要酶，但其在柴胡皂苷合成中的作用研究甚少。本研究通过代谢组学和转录组学分析筛选出的β-AS基因BcBAS1(Accession No.ON890382)克隆并在大肠杆菌和毕氏酵母中进行了异源表达功能表征，并对其亚细胞定位和表达模式进行了研究。重组BcBAS1蛋白分子量约为87 kDa，该蛋白可催化SSs前体β-amyrin的生成。此外，BcBAS1位于细胞质中，4种基因型在4种组织中的相对表达量与SSa和SSd含量呈正相关。我们的结果表明，BcBAS1是一个β-AS基因，可能在柴草皂苷的生物合成和调控中发挥重要作用。本研究揭示了β-AS基因在SSs合成中的作用，为SSs的代谢工程提供了新的思路。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.