Molecular and biochemical characterization of squalene synthase from Siraitia grosvenorii

Su, Heling; Liu, Yongming; Xiao, Yalun; Yue-jin, Tan; Gu, Yunyan; Liang, Bin; Huang, Honglan

doi:10.1007/s10529-017-2328-z

Cited by 7 publications

(4 citation statements)

References 15 publications

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“…Thirdly, the alpha helices of the monomeric SQS protein form a cave-like active center and transmembrane domain(s) at the C-terminus. In recent years, the transmembrane regions have been identified by bioinformatics approaches in a variety of species, such as Siraitia grosvenorii , wintersweets and Cucurbitaceae family plants [36,44,45]. The enzymatically active center folded by helices supplies an interacting surface with SQS substrate FPP via hydrogen bonds and hydrophobic interactions (Figure 3).…”

Section: Discussionmentioning

confidence: 99%

“…Expression analysis indicated that the ubiquitous MsSQS was expressed preferentially in roots (Figure 6). The root-preferred pattern was observed for GmSQS1 in soybean [11], SgSQS in Siraitia grosvenorii [44], HsSQS1 in Huperzia serrata [46] and TwSQS in traditional Chinese medicinal plant Tripterygium wilfordii [37]. Some plants, such as Withania somnifera [47], Betula platyphylla [15] and Arabidopsis [22], displayed a leaf-predominant pattern, suggesting that the spatial and temporal expression patterns of SQS genes vary greatly in different plants.…”

Section: Discussionmentioning

confidence: 99%

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Molecular Cloning and Functional Identification of a Squalene Synthase Encoding Gene from Alfalfa (Medicago sativa L.)

Zhang

et al. 2019

IJMS

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The quality of alfalfa, a main legume forage worldwide, is of great importance for the dairy industry and is affected by the content of triterpene saponins. These natural terpenoid products of triterpene aglycones are catalyzed by squalene synthase (SQS), a highly conserved enzyme present in eukaryotes. However, there is scare information on alfalfa SQS. Here, an open reading frame (ORF) of SQS was cloned from alfalfa. Sequence analysis showed MsSQS had the same exon/intron composition and shared high homology with its orthologs. Bioinformatic analysis revealed the deduced MsSQS had two transmembrane domains. When transiently expressed, GFP-MsSQS fusion protein was localized on the plasma membrane of onion epidermal cells. Removal of the C-terminal transmembrane domain of MsSQS improved solubility in Escherichia coli. MsSQS was preferably expressed in roots, followed by leaves and stems. MeJA treatment induced MsSQS expression and increased the content of total saponins. Overexpression of MsSQS in alfalfa led to the accumulation of total saponins, suggesting a correlation between MsSQS expression level with saponins content. Therefore, MsSQS is a canonical squalene synthase and contributes to saponin synthesis in alfalfa. This study provides a key candidate gene for genetic manipulation of the synthesis of triterpene saponins, which impact both plant and animal health.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Molecular Cloning and Functional Identification of a Squalene Synthase Encoding Gene from Alfalfa (Medicago sativa L.)

Zhang

et al. 2019

IJMS

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“…An HPLC analysis showed SS oil contained 12.5% squalene ( 9 , 13 ). Squalene has the functions of body building and antifatigue and can treat liver diseases ( 14 ). As a high-quality plant protein resource, its emulsifying and foaming properties are better than those of soybean protein isolate ( 15 ).…”

Section: Introductionmentioning

confidence: 99%

Effects of Siraitia grosvenorii seed flour on the properties and quality of steamed bread

Zhou,

Guo,

Zhang

et al. 2023

Front. Nutr.

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Siraitia grosvenorii seeds are rich in abundant active compounds beneficial to human health. To clarify the digestion characteristics of Siraitia grosvenorii seed flour (SSF) and promote the use of SSF in the processing of functional staple foods, SSF was prepared, its composition and physicochemical properties were studied, and the processing characteristics of SSF-wheat flour were systematically investigated. The results showed that the torque curve and other parameters of the dough were significantly affected by the amount of SSF added. With the increase of SSF proportion, the water absorption showed an increasing trend, while the degree of protein weakening first weakened and then enhanced. At 20% SSF, the dough was more resistant to kneading. In response to an increase in SSF, the L* value decreased significantly, and the a* and b* values increased gradually, while the specific volume decreased gradually. Additionally, the hardness, adhesiveness, and chewiness of the bread enhanced gradually, while its elasticity, cohesiveness, and resilience decreased gradually. After the addition of 30% SSF, the inner tissue of steamed bread was more delicate. With an increase in SSF proportion, the predicted glycemic index (pGI) of steamed bread weakened markedly. Overall, these results showed that SSF, as a kind of food ingredient with hypoglycemic activity, can be used in the production of new functional steamed bread products. This study provides basic research data for the development of products containing S. grosvenorii seed.

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“…Squalene is thought to be the initial substrate and precursor for triterpenoid and sterol biosynthesis. SQE has been generally recognized as the common rate-limiting enzyme in the common pathway from mevalonate (MVA) and methylerythritol phosphate (MEP) pathways, catalyzing squalene to 2,3-oxidosqualene [17,18]. In S. grosvenorii , the initial step in the biosynthesis of cucurbitane-type mogrosides is the cyclization of 2,3-oxidosqualene to form the triterpenoid skeleton of cucurbitadienol, which is catalyzed by CS.…”

Section: Introductionmentioning

confidence: 99%

Identification of a Novel Specific Cucurbitadienol Synthase Allele in Siraitia grosvenorii Correlates with High Catalytic Efficiency

Qiao

Luo

et al. 2019

Molecules

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Mogrosides, the main bioactive compounds isolated from the fruits of Siraitia grosvenorii, are a group of cucurbitane-type triterpenoid glycosides that exhibit a wide range of notable biological activities and are commercially available worldwide as natural sweeteners. However, the extraction cost is high due to their relatively low contents in plants. Therefore, molecular breeding needs to be achieved when conventional plant breeding can hardly improve the quality so far. In this study, the levels of 21 active mogrosides and two precursors in 15 S. grosvenorii varieties were determined by HPLC-MS/MS and GC-MS, respectively. The results showed that the variations in mogroside V content may be caused by the accumulation of cucurbitadienol. Furthermore, a total of four wild-type cucurbitadienol synthase protein variants (50R573L, 50C573L, 50R573Q, and 50C573Q) based on two missense mutation single nucleotide polymorphism (SNP) sites were discovered. An in vitro enzyme reaction analysis indicated that 50R573L had the highest activity, with a specific activity of 10.24 nmol min−1 mg−1. In addition, a site-directed mutant, namely, 50K573L, showed a 33% enhancement of catalytic efficiency compared to wild-type 50R573L. Our findings identify a novel cucurbitadienol synthase allele correlates with high catalytic efficiency. These results are valuable for the molecular breeding of luohanguo.

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Molecular and biochemical characterization of squalene synthase from Siraitia grosvenorii

Abstract: SgSQS gene was cloned, and the molecular structure and biochemical function of SgSQS were characterized.

Cited by 7 publications

References 15 publications

Molecular Cloning and Functional Identification of a Squalene Synthase Encoding Gene from Alfalfa (Medicago sativa L.)

Molecular Cloning and Functional Identification of a Squalene Synthase Encoding Gene from Alfalfa (Medicago sativa L.)

Effects of Siraitia grosvenorii seed flour on the properties and quality of steamed bread

Identification of a Novel Specific Cucurbitadienol Synthase Allele in Siraitia grosvenorii Correlates with High Catalytic Efficiency

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