Phosphate → Steviol

HRID 223343

Reaction equation

HRID 223343 reaction equation
C[C@H]1O[C@@H]([O:8][C@:9]23[C:24](=[CH2:25])[CH2:23][C@:21]4([CH2:22]2)[C@H:12]([C@@:13]2([CH3:41])[C@H:18]([CH2:19][CH2:20]4)[C@@:17]([C:27]([O:29][C@@H]4O[C@H](CO)[C@@H](O)[C@H](O)[C@H]4O)=[O:28])([CH3:26])[CH2:16][CH2:15][CH2:14]2)[CH2:11][CH2:10]3)[C@@H](O[C@@H]2O[C@H](CO)[C@@H](O)[C@H](O[C@@H]3O[C@H](CO)[C@@H](O)[C@H](O)[C@H]3O)[C@H]2O)[C@@H](O)[C@@H]1O.[K].P([O-])([O-])([O-])=O.SC[C@H]([C@@H](CS)O)O.O[C@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O>OP([O-])(O)=O.[K+].CC#N.C(Cl)(Cl)Cl.CO.O>[CH3:41][C@:13]12[C@@H:12]3[CH2:11][CH2:10][C@@:9]4([OH:8])[C:24]([CH2:23][C@@:21]3([CH2:22]4)[CH2:20][CH2:19][C@@H:18]1[C@@:17]([C:27]([OH:29])=[O:28])([CH3:26])[CH2:16][CH2:15][CH2:14]2)=[CH2:25]

Participants

9

Reactant 5

HCID 1061PhosphateHCID 79025Alpha-D-GlucoseHCID 446094D-1,4-DithiothreitolHCID 5462222PotassiumHCID 44564216[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl] (1R,4S,5R,9S,10R,13S)-13-[(2S,3S,4S,5S,6R)-3-[(2S,3R,4S,5R,6R)-3,5-dihydroxy-6-(hydroxymethyl)-4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyoxan-2-yl]oxy-4,5-dihydroxy-6-methyloxan-2-yl]oxy-5,9-dimethyl-14-methylidenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate2×

Product 1

HCID 452967Steviol

Reagents, catalysts & solvents 3

HCID 6342AcetonitrileSolventHCID 516951Monobasic potassium phosphateSolventHCID 21866399Chloroform methanol waterSolvent

Procedure

Rebaudioside C, 3-sulfopropyl ester, potassium salt prepared as in Part B. was incubated anaerobically for three days at 37° C. with 5% fresh rat cecal contents at concentrations of 0.25, 0.5, and 1.0 mg/ml, in sterile Krebs-Ringer 0.25 M phosphate buffer (pH 7.4) containing 0.25 mg/ml dithiothreitol and 0.25 mg/ml α-D-glucose. TLC [silica gel F-254; CHCl3 :CH3OH:H2O (15:10:2)] and HPLC [30 cm C-18 on μ-Bondapak; 15 min linear gradient of 10-40% CH3CN in 0.005 M KH2PO4 (pH 3.45); 200 nm] analysis showed all the rebaudioside C, 4-sulfopropyl ester (Rf =0.42; tR =14.0 min) to have been consumed within 24 h to yield apparently only the sulfopropyl ester of steviol, (Rf =0.63; tR =16.3 min). No steviol (Rf =0.95; tR =31.3 min) was detected. After 3 days the bacterial cells were sedimented by centrifugation. The sediment was extracted (THF), as was the lyophilized supernatant. HPLC analysis for steviol of the two THF extracts showed none to be detectable. With a detection limit of 0.05 μg, as little as 0.03 and 0.13 percent degradation to steviol could have been detected for the sediment and supernatant fractions, respectively.

Workup

  1. customto have been consumed within 24 h

Sources & evidence

ORD record
ord-e88ee83601aa49e4a59a7ec3e76ed946
Source dataset
uspto-grants-1982_10
Patent number
US04353889

Linked notes

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