Method for preparing ginseng saponin F2 by enzymatic hydrolysis of ginseng saponin Rb1
A technology of ginsenoside and enzymatic hydrolysis, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of cumbersome separation and complex components of reaction products, and achieve low process cost and benefit large quantities The effect of mild preparation and reaction conditions
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Embodiment 1
[0014] Cultivate Fusarium ECU2042 in Chase's medium, medium composition: sucrose 30g / l, NaNO 3 3.0g / l, K 2 HPO 4 1.0g / l, KCl 0.5g / l, MgSO 4 ·7H 2 O 0.5g / l, FeSO 4 ·7H 2 O 0.01 g / l.
[0015] Seed liquid is cultivated in 30 ℃, 180rpm shaking table after 18 hours, inserts in the 500ml shaking flask (filling liquid volume 100ml) by 4% inoculum size, after cultivating 72 hours under the same condition, filter and collect thalli.
[0016] Use the method of grinding to crush the cells, centrifuge the cell crushing liquid with a high-speed refrigerated centrifuge (12,000rpm, 30min), collect the supernatant, slowly add an equal volume of refrigerated acetone under ice bath conditions, continue to stir for 30 minutes, and then use a high-speed refrigerated centrifuge After centrifugation, the precipitate is vacuum freeze-dried to obtain the Fusarium crude enzyme preparation.
Embodiment 2
[0018] In the 100ml grinding mouth Erlenmeyer flask, add successively the crude enzyme that 600mg makes in the embodiment 1, 60mg ginsenoside Rb 1 , then add 30ml of acetate buffer (NaAc / HAc, 0.2M, pH 5.0), plug the ground plug tightly, seal it with raw material tape, react on a rotary shaker at 200rpm at 50°C for 72 hours, and take a sample for HPLC Analysis (detection: UV 203nm; mobile phase: methanol / water=85 / 15, v / v), the conversion rate was about 30.5%.
Embodiment 3
[0020] In the 100ml grinding mouth Erlenmeyer flask, add successively the crude enzyme that 300mg makes in the embodiment 1, 60mg ginsenoside Rb 1 , then add 30ml of acetate buffer solution (NaAc / HAc, 0.2M, pH 5.0), plug the ground plug tightly, seal it with raw material tape, react on a rotary shaker at 200rpm at 50°C for 72 hours, and then add 300mg Crude enzyme, continue to react for 48 hours, add 30ml of n-butanol to extract 4 times respectively, collect n-butanol phase, concentrate by rotary evaporation, and separate through preparative chromatographic column (Venusil XBP C18) to obtain 16mg of ginsenoside F 2 , the conversion rate is about 43.2%, and the yield is 87%, the prepared ginsenoside F 2 The purity is ≥95%.
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