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Cloning and Functional Characterization of a Squalene Synthase from Paris polyphylla var. yunnanensis
Gao,Jian-Xiong; Chen,Yue-Gui; Li,De-Sen; Lin,Liang; Liu,Yan; Li,Sheng-Hong
2021
Source PublicationCHEMISTRY & BIODIVERSITY
ISSN1612-1872
Volume18Issue:7
AbstractParis polyphylla Smith var. yunnanensis (Franch.) Hand. - Mazz. is a precious traditional Chinese medicine, and steroidal saponins are its major bioactive constituents possessing extensive biological activities. Squalene synthase (SQS) catalyzes the first dedicated step converting two molecular of farnesyl diphosphate (FDP) into squalene, a key intermediate in the biosynthetic pathway of steroidal saponins. In this study, a squalene synthase gene (PpSQS1) was cloned and functionally characterized from P. polyphylla var. yunnanensis, representing the first identified SQS from the genus Paris. The open reading frame of PpSQS1 is 1239 bp, which encodes a protein of 412 amino acids showing high similarity to those of other plant SQSs. Expression of PpSQS1 in Escherichia coli resulted in production of soluble recombinant proteins. Gas chromatography-mass spectrometry analysis showed that the purified recombinant PpSQS1 protein could produce squalene using FDP as a substrate in the in vitro enzymatic assay. qRT-PCR analysis indicated that PpSQS1 was highly expressed in rhizomes, consistent with the dominant accumulation of steroidal saponins there, suggesting that PpSQS1 is likely involved in the biosynthesis of steroidal saponins in the plant. The findings lay a foundation for further investigation on the biosynthesis and regulation of steroidal saponins, and also provide an alternative gene for manipulation of steroid production using synthetic biology.
KeywordParis polyphylla var Yunnanensis squalene synthase steroidal saponins squalene in vitro enzymatic assay MOLECULAR-CLONING SAPONINS EXPRESSION GENE GLYCOSIDES PLANTS SQS
DOI10.1002/cbdv.202100342
WOS IDWOS:000662745000001
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Document Type期刊论文
Identifierhttp://ir.kib.ac.cn/handle/151853/73449
Collection中国科学院昆明植物研究所
Affiliation1.Chinese Acad Sci, Kunming Inst Bot, State Key Lab Phytochem & Plant Resources West Ch, Kunming 650201, Yunnan, Peoples R China
2.Chinese Acad Sci, Kunming Inst Bot, Yunnan Key Lab Nat Med Chem, Kunming 650201, Yunnan, Peoples R China
3.Chinese Acad Sci, Kunming Inst Bot, Germplasm Bank Wild Species, Kunming 650201, Yunnan, Peoples R China
4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Gao,Jian-Xiong,Chen,Yue-Gui,Li,De-Sen,et al. Cloning and Functional Characterization of a Squalene Synthase from Paris polyphylla var. yunnanensis[J]. CHEMISTRY & BIODIVERSITY,2021,18(7).
APA Gao,Jian-Xiong,Chen,Yue-Gui,Li,De-Sen,Lin,Liang,Liu,Yan,&Li,Sheng-Hong.(2021).Cloning and Functional Characterization of a Squalene Synthase from Paris polyphylla var. yunnanensis.CHEMISTRY & BIODIVERSITY,18(7).
MLA Gao,Jian-Xiong,et al."Cloning and Functional Characterization of a Squalene Synthase from Paris polyphylla var. yunnanensis".CHEMISTRY & BIODIVERSITY 18.7(2021).
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