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Structure and Location Studies on Key Enzymes in Saponins Biosynthesis of Panax notoginseng

by Pengguo Xia 1,2,*, Yujie Zheng 1 and Zongsuo Liang 1,*
1
Laboratory of Plant Secondary Metabolism and Regulation of Zhejiang Province, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China
2
State Key Laboratory of Membrane Biology, Innovation Center for Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(24), 6121; https://doi.org/10.3390/ijms20246121
Received: 17 October 2019 / Revised: 19 November 2019 / Accepted: 3 December 2019 / Published: 4 December 2019
(This article belongs to the Section Molecular Plant Sciences)
Panax notoginseng is one of the most widely used traditional herbs for the treatment of various diseases, in which saponins were the main active components. At present, the research of P. notoginseng mainly focused on the discovery of new compounds and pharmacology. However, there were few studies on the molecular mechanism of the synthesis of secondary metabolites of P. notoginseng. In our study, four coding sequences (CDS) encoding the key enzymes involved in saponin biosynthesis were cloned, namely farnesyl diphosphate synthase (FPS), squalene synthase (SS), squalene epoxidase (SE), and dammarenediol-II synthase (DS), which contained open reading frame (ORF) of 1029 bp, 1248 bp, 1614 bp, and 2310 bp, and coded 342, 415, 537, and 769 amino acids, respectively. At the same time, their domains, secondary structures, three-dimensional structures, and phylogenetics trees were analyzed by kinds of bioinformatics tools. Their phylogenetics relationships were also analyzed. In addition, GFP (Green fluorescent protein) fusion genes were constructed by the plasmid transformation system to determine the subcellular localization. The results of subcellular localization showed that FPS, SE, and DS were mainly located in cytomembrane and its surrounding, while SS was located both in cytoplasm and cytomembrane. Our findings provided data demonstrating the expression patterns of genes involved in saponin biosynthesis and would facilitate efforts to further elucidate the biosynthesis of the bioactive components in P. notoginseng.
Keywords: Panax notoginseng; saponins biosynthesis; key enzymes; subcellular localization Panax notoginseng; saponins biosynthesis; key enzymes; subcellular localization
MDPI and ACS Style

Xia, P.; Zheng, Y.; Liang, Z. Structure and Location Studies on Key Enzymes in Saponins Biosynthesis of Panax notoginseng. Int. J. Mol. Sci. 2019, 20, 6121.

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