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Turkish Journal of Agriculture and Forestry

Abstract

Ilex cornuta (I. cornuta) is a traditional Chinese medicine that is rich in various secondary metabolites with pharmacological activities, among which triterpenoid saponins are one of the main active components. Squalene epoxidase (SE) is a key regulatory enzyme in the biosynthesis of triterpenoids. In this study, we isolated a squalene epoxidase (IcSE1) from I. cornuta and analyzed its molecular characteristics and functions. The results indicated that the total length of the IcSE1 sequence was 1942 bp, which includes a 1638 bp open reading frame (ORF) that encodes 545 amino acids. The theoretical isoelectric point (pI) and molecular weight of the IcSE1 protein were found to be 7.51 and 59.8 kDa, respectively. This protein was determined to be hydrophobic and is predicted to localize to the plasma membrane. Phylogenetic analysis revealed that IcSE1 is closely related to the SE protein of Panax vietnamensis. Tissue expression pattern analysis demonstrated that IcSE1 expression was highest in roots, followed by leaves and male flowers, with the lowest expression observed in female flowers. In this study, we successfully constructed a prokaryotic expression system for the IcSE1 gene, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) results confirmed that the size of the expressed protein was consistent with the predicted values. Furthermore, we successfully established a yeast expression system for the IcSE1 gene. Analysis of the yeast expression products using high-performance liquid chromatography (HPLC) showed that the total amount of triterpene saponins in the transgenic yeast was 1.2 times higher than that of the control. These findings provide a theoretical basis for investigating the molecular mechanisms underlying the biosynthesis of triterpenoid saponins from I. cornuta.

Author ORCID Identifier

XIAOXI ZHANG: 0000-0001-8042-3393

LI ZHU: 0009-0004-5695-5241

WEIWEI ZHANG: 0000-0003-0800-4797

FENG XU: 0000-0003-3212-6284

JIABAO YE: 0000-0002-2076-8880

YONGLING LIAO: 0009-0008-1580-3080

DOI

10.55730/1300-011X.3258

Keywords

Ilex cornuta, squalene epoxidase, triterpenoids saponins, procaryotic expression, yeast expression

First Page

194

Last Page

204

Creative Commons License

Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.

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