Rebaudioside D

Rebaudioside D, a glycoside of the diterpene steviol is a minor constituent found in leaves of Stevia rebaudiana. It is considered non-caloric and is about 200–220 times sweeter than sucrose.
Rebaudioside D is a natural non-caloric sweetener. It is a steviol glycoside that has been found in S. rebaudiana leaves. Rebaudioside D, similarly to rebaudioside A and rebaudioside C, is metabolized by gut microbiota to steviol, a compound whose safety is widely studied.

Rebaudioside D is a steviol glycoside used as a nutritive sweetener and may be used as a replacement for sugar in foods and beverages. Useful in the treatment of blood glucose sensitive diseases such as diabetes, high blood pressure, and obesity.
It may be used as internal standard to optimize the desorption electrospray ionization (DESI) mass spectrometry during the analysis of Stevia leaves for diterpene glycosides.

The rebaudioside D-rich steviol glycoside preparation (~95% rebaudioside D) is produced via an enzymatic conversion process using a strain of Pichia pastoris that has been genetically modified to express UDPglucosyltransferase EUGTll. In the first stage of manufacturing, a steviol glycoside primary extract from the leaves of 5. rebaudiana Bertoni containing 55±5% of rebaudioside A is produced according to the methodology outlined in the Chemical and Technical Assessment (CTA) published by the Food and Agriculture Organization (FAO)/JECFA for steviol glycosides (FAQ, 2016). In the next step, the steviol glycoside primary extract is further purified to a purity of ~95% rebaudioside A through crystallization, also consistent with the CTA methodology. In the third stage, the P. pastoris production strain is subjected to a fermentation step to express the UGT-glucosyltransferase EUGTll, which is used to catalyze the conversion of the high-purity rebaudioside A extracted from 5. rebaudiana Bertoni to rebaudioside D. In the last stage, the rebaudioside D solution is purified and concentrated according to the methodology described in the steviol glycoside CTA, yielding a final product that contains ~95% rebaudioside D.
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