Journal
FOOD CHEMISTRY
Volume 259, Issue -, Pages 286-291Publisher
ELSEVIER SCI LTD
DOI: 10.1016/j.foodchem.2018.03.126
Keywords
Stevia rebaudiana; UDP-glycosyltransferase; UGTSL2; Rebaudioside D; Rebaudioside M2
Funding
- NSFC - China [21106068]
- Open Fund Program of the Yichang Key Laboratory of Biocatalysis - China [2015NP01]
- Provincial Key R&D Plan of Jiangsu - China [BE2017703]
- TAPP - China
- Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture - China
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Steviol glycosides from Stevia rebaudiana leaves are used in stevia-based sweeteners for their intense sweetness and low calories. Rebaudioside D is present in leaves in minute quantities (similar to 0.4-0.5% w/w total dry weight), but it is similar to 350 times sweeter than sucrose, and sweeter than the more abundant rebaudioside A and stevioside. In the present study, pathways for rebaudioside D synthesis and UDP-glucose recycling were developed by coupling recombinant UDP-glucosyltransferase UGTSL2 from Solanum lycopersicum and sucrose synthase StSUS1 from Solanum tuberosum. Reaction parameters, including substrate ratio, sucrose concentration, temperature, crude extract concentration, and reaction time, were evaluated, and 17.4 g/l of rebaudioside D (yield = 74.6%) was obtained from 20 g/l of rebaudioside A after 20 h, using UDP or UDP-glucose in recombinant cell crude extracts. Extending the reaction time generated rebaudioside M2 from further glycosylation of rebaudioside D. K-m values for UGTSL2 indicated a higher affinity for rebaudioside D than for rebaudioside A.
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