Rebaudioside A synthesis
- Product Name:Rebaudioside A
- CAS Number:58543-16-1
- Molecular formula:C44H70O23
- Molecular Weight:967.01
57817-89-7
57-50-1
58543-16-1
A new structure based on (2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl((4R,4aS,6aR,9S,11aR,11bS)-9-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R )-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy-4,11b-dimethyl-8-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylic acid ester and sucrose were used as raw materials, and the general procedure for the synthesis of rebaudioside A was as follows: firstly, the expression in E. coli of pEUGT- SUS vector in E. coli, followed by collecting the cells and washing them twice with potassium phosphate buffer (100 mM, pH 7.2). Next, the cells were broken by sonication in an ice-water bath and centrifuged at 4°C to remove cellular debris to obtain a lysate supernatant containing soluble proteins. This supernatant is the crude extract. The total protein concentration of the crude extract was determined by the Bradford method using bovine serum albumin (BSA) as a standard. Finally, rebaudioside A was synthesized by reacting the substrate acaricide with an equal volume of the crude extract in potassium phosphate buffer (100 mM, pH 7.2) containing different concentrations of UDP-glucose or UDP, sucrose, and 3 mM MgCl2 at 30 °C.
57817-89-7
586 suppliers
$5.00/1g
57-50-1
630 suppliers
$5.00/10g
58543-16-1
274 suppliers
$5.00/25mg
Yield:58543-16-1 78%
Reaction Conditions:
with Arabidopsis thaliana sucrose synthase gene;UDP-glucosyltransferase from Stevia rebaudiana, recombinant;magnesium chloride in aq. phosphate buffer; pH=7.2 at 30; for 30 h;Enzymatic reaction;Time;
Steps:
Enzymatic synthesis of rebaudioside A with UGT76G1 and AtSUS1
After pEUGT-SUS expressionin E. coli, the cells were harvested and washed twice in potassium phosphate buffer (100 mM, pH 7.2). A lysate containing soluble proteins was obtained by sonication on an ice-water bath and subsequent clarification by centrifugation at 4 °C. The supernatant was used as the crude extract. Total protein concentrations were measured by the Bradford assay using bovine serum albumin (BSA) as a standard.23) The substratestevioside was incubated with aliquots of the crude extract at 30 °C in a reaction mixture containing varying concentrations of UDP-glucose or UDP, sucrose, and 3 mM MgCl2 in potassium phosphate buffer (100 mM, pH 7.2) for synthesis of rebaudioside A.
References:
Wang, Yu;Chen, Liangliang;Li, Yan;Li, Yangyang;Yan, Ming;Chen, Kequan;Hao, Ning;Xu, Lin [Bioscience, Biotechnology and Biochemistry,2016,vol. 80,# 1,p. 67 - 73]
57817-89-7
586 suppliers
$5.00/1g
58543-16-1
274 suppliers
$5.00/25mg
133-89-1
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58543-16-1
274 suppliers
$5.00/25mg
57-50-1
630 suppliers
$5.00/10g
58543-16-1
274 suppliers
$5.00/25mg