Method for enzymatic synthesis of gadol glycoside through separation and coupling
A salidroside and enzymatic synthesis technology, applied in chemical recovery, fermentation, etc., can solve problems such as efficiency impact, and achieve high added value, high purity, and low cost effects
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Embodiment 1
[0027] Preparation of apple seed powder: take 1kg of apple seed waste from the preserved fruit factory, grind or mash it into powder, wash with ethyl acid 3 times (300ml×3), then wash with 300ml acetone once, drain and place Dry in a vacuum drying oven to obtain a total of 350g of dry apple seed powder, which is placed in a refrigerator at 4°C for later use.
Embodiment 2
[0029] Add 49mg glucose to 0.1mL potassium phosphate buffer (50mmol / L, pH6.5) and heat to dissolve. The final concentration of sugar in 1mL reaction system is 0.25mol / L; 0.383g p-hydroxyphenethyl alcohol is added to 0.9mL tert-butyl Heat and dissolve in alcohol. The final concentration of alcohol in 1mL reaction system is 2.5mol / L. After cooling, add 15mg (1.52U) of crude apple seed enzyme powder. After shaking and reacting at 50℃ for 192 hours, salidroside is analyzed and determined. The final yield was 28.2%.
Embodiment 3
[0031] Add 250mg apple seed powder, 272mg glucose, 0.5ml phosphate buffer (70mmol / L Na 2 HPO 4 -KH 2 PO 4 , PH6.0), 2.28g p-hydroxyphenethyl alcohol, 5.0ml dioxane, stopper the ground bottle stopper tightly, and seal with raw material tape, react at 45℃, 160rpm shaker for 120h.
[0032] After the reaction, 10ml of methanol was added to remove the enzyme by filtration. The filtered enzyme was washed with methanol (20ml×3). The filtrate was dried by adding anhydrous sodium sulfate, and the solvent was removed by rotary evaporation. Flash column chromatography (ethyl acetate: methanol = 13:1), 66 mg of salidroside was obtained, and the yield relative to glucose was 15.8%.
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