Phosphate → Steviol

HRID 1262901

Reaction equation

HRID 1262901 reaction equation
[CH3:1][C@:2]12[C@@H:11]3[CH2:12][CH2:13][C@@:14]4([O:19][C@@H]5O[C@@H](CO)[C@@H](O)[C@H](O)[C@@H]5O[C@@H]5O[C@@H](CO)[C@@H](O)[C@H](O)[C@@H]5O)[C:16]([CH2:18][C@@:10]3([CH2:15]4)[CH2:9][CH2:8][C@@H:7]1[C@@:6]([C:43]([O-:45])=[O:44])([CH3:42])[CH2:5][CH2:4][CH2:3]2)=[CH2:17].[Na+].[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>[CH3:1][C@:2]12[C@@H:11]3[CH2:12][CH2:13][C@@:14]4([OH:19])[C:16]([CH2:18][C@@:10]3([CH2:15]4)[CH2:9][CH2:8][C@@H:7]1[C@@:6]([C:43]([OH:45])=[O:44])([CH3:42])[CH2:5][CH2:4][CH2:3]2)=[CH2:17]

Participants

9

Reactant 5

HCID 1061PhosphateHCID 79025Alpha-D-GlucoseHCID 446094D-1,4-DithiothreitolHCID 5462222PotassiumHCID 73352876HCID 73352876, compound

Product 1

HCID 452967Steviol

Reagents, catalysts & solvents 3

HCID 6342AcetonitrileSolventHCID 516951Monobasic potassium phosphateSolventHCID 21866399Chloroform methanol waterSolvent

Procedure

Steviolbioside, 3-sulfopropyl ester, potassium salt prepared in Part B. was incubated anaerobically for three days at 37° C. with 5 wt% fresh rat cecal contents, at concentrations of 0.25, 0.5, and 1.0 mg/ml in sterile Krebs-Ringer 0.25M 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 0.005M KH2PO4 (pH 3.45); 200 nm] analysis showed all the initial 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 supernatant, after lyophilization. HPLC analysis for steviol of the two THF extracts showed none to be detectable. With a detection limit of 0.05 g, 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. customtR =14.0 min) to have been consumed within 24 h

Sources & evidence

ORD record
ord-2348d9a2efe5469ba8a16b193cc98e61
Source dataset
uspto-grants-1984_06
Patent number
US04454290

Linked notes

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