Separation of rebaudioside D from steviol glycoside by frontal chromatography

The biosynthesis of steviol glycosides is affected by both genetic and environmental factors. To evaluate the influence of total daily solar radiation or daily light integral (DLI) under long-day conditions on steviol glycoside concentration, we grew Stevia rebaudiana under ambient irradiance or varying levels of shading at different times of the year in both greenhouse and field environments, resulting in DLIs ranging from 3.55 to 20.31 mol·m−2·d−1 in the greenhouse and 10.32 to 39.7 mol·m−2·d−1 in the field. Total steviol glycoside concentration of selected leaves from greenhouse-grown plants increased as DLI increased up to ca. 10 mol·m−2·d−1, remaining constant with further increases in DLI, and was similar across the range of DLIs evaluated in the field. DLI influenced both the concentration and the relative proportions of specific steviol glycosides. Rebaudioside A concentration increased as DLI increased from 3.55 to 8.53 mol·m−2·d−1, remaining similar with further increases in DLI. Rebaudioside D and stevioside concentration of selected leaves from field-grown plants decreased by 22% and 13%, respectively, as DLI increased from 10.32 to 39.7 mol·m−2·d−1, while rebaudioside A and M concentrations remained similar across this DLI range. Collectively, these results indicate that the greatest influence of DLI on steviol glycoside concentration occurs under relatively low DLIs (<10 mol·m−2·d−1). However, higher DLIs can significantly affect the synthesis of minor glycosides of increasing commercial importance including rebaudioside D.

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Isolation and Structure Elucidation of Rebaudioside D2 from Bioconversion Reaction of Rebaudioside A to Rebaudioside D

Natural Product Communications ◽

10.1177/1934578x1400900819 ◽

2014 ◽

Vol 9 (8) ◽

pp. 1934578X1400900 ◽

Cited By ~ 3

Author(s):

Indra Prakash ◽

Cynthia Bunders ◽

Krishna P. Devkota ◽

Romila D. Charan ◽

Catherine Ramirez ◽

...

Keyword(s):

Spectral Data ◽

Structure Elucidation ◽

Rebaudioside A ◽

Mass Spectral Data ◽

Complete Structure ◽

Mass Spectral ◽

Steviol Glycoside ◽

Extensive Analysis ◽

Rebaudioside D

We report the isolation and complete structure of an isomer of rebaudioside D, known as rebaudioside D2. This novel steviol glycoside was isolated from a bioconversion reaction of rebaudioside A to rebaudioside D. Rebaudioside D2 possesses a relatively rare 1→6 sugar linkage, which was discovered by extensive analysis of NMR (1H, 13C, COSY, HSQC-DEPT, HMBC, 1D TOCSY and NOESY) and mass spectral data.

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Development and Validation of a Reversed-Phase High-Performance Liquid Chromatography Method for Routine Identification and Purity Assessment of High-Purity Steviol Glycoside Sweeteners

Journal of Agricultural and Food Chemistry ◽

10.1021/jf4047557 ◽

2014 ◽

Vol 62 (6) ◽

pp. 1384-1389 ◽

Cited By ~ 5

Author(s):

Tsion Bililign ◽

Jeffrey C. Moore ◽

Shane Tan ◽

Allan T. Leeks

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Purity ◽

High Performance ◽

Reversed Phase ◽

Chromatography Method ◽

Steviol Glycoside ◽

Routine Identification ◽

Liquid Chromatography Method ◽

Development And Validation

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Daminozide Enhances Vigor and Steviol Glycoside Yield of Stevia (Stevia rebaudiana Bert.) Propagated in the Temporary Immersion Bioreactor

10.21203/rs.3.rs-1223930/v1 ◽

2022 ◽

Author(s):

Rizka Tamania Saptari ◽

Rizkita Rachmi Esyanti ◽

Riza Arief Putranto

Keyword(s):

Stevia Rebaudiana ◽

Feedback Regulation ◽

Optimum Concentration ◽

Temporary Immersion Bioreactor ◽

Temporary Immersion ◽

Steviol Glycoside ◽

Kaurenoic Acid ◽

Biosynthesis Gene ◽

Inhibitor Treatment ◽

Ga Biosynthesis

Abstract Stevia (Stevia rebaudiana Bertoni) contains sweet compound widely used as natural sweetener, steviol glycoside (SG). SG is a diterpenoid secondary metabolite synthesized from ent-kaurenoic acid, the same precursor of Gibberellin (GA). Therefore, in this study, a GA inhibitor, Daminozide (0, 10, 20 ppm) was used to block ent-kaurenoic acid conversion towards GA synthesis in attempt to increase SG content of stevia propagated in Temporary Immersion Bioreactor (TIB). Daminozide in 10 mg/L was observed to be the optimum concentration which increased biomass weight and SG content (stevioside and rebaudioside A) up to 40%. The treatment also increased transcripts accumulation of genes enrolled in SG biosynthesis, such as SrKA13H, SrUGT85C2, and SrUGT76G1, indicating SG pathway become more active due to the inhibition of GA pathway. Furthermore, the inhibition of GA was also indicated by the upregulated expression of GA biosynthesis gene (GA3ox) as the result of feedback regulation, and the downregulated expression of GA catabolism gene (GA2ox2) as the result of feed-forward regulation caused by inhibitor treatment.

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