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Method for preparing hypoglycosyl dioscin by enzyme method hydrolyzing yam saponin glycosyl

A technology for diosgenin and saponin sugar, which is applied in the field of preparing low-glycosylated diosgenin, can solve the problems of large saponin breakage and pollution, and achieve the effects of low hemolysis, high curative effect and high conversion rate.

Active Publication Date: 2004-03-24
SUNFLOWER PHARM GRP (TIANJIN) DRUG RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to obtain low-glycoside saponins and aglycones, acid-base hydrolysis has been reported (Guo Wensong et al., Guizhou Chemical Industry, 1996, Phase II, 14-17), but the damage to saponins is large and the pollution is serious

Method used

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  • Method for preparing hypoglycosyl dioscin by enzyme method hydrolyzing yam saponin glycosyl

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] a. Take thermoaerobic bacteria Bacillus sp.JF 2 Bacteria (document 1) were cultured with ventilation at a temperature of 60°C for 30-40 hours in a culture medium containing 3% corn flour, 0.2% enzyme-producing inducer - yam extract (dry matter), 0.01% MgSO4, and centrifuged to remove Bacteria, adding 65% saturated ammonium sulfate to the supernatant (containing the mixed solution of enzyme) to precipitate the enzyme protein, dialyzing with 0.02M, pH5 acetic acid buffer, centrifuging to remove slag, freeze-drying to obtain saponinase;

[0033] b. Get 2 grams of the above-mentioned saponin enzyme, 30 grams of dioscin 1, 1500 milliliters of acetic acid buffer (0.2M, pH5.0) and 150 milliliters of ethanol and mix evenly, so that 30 grams of dioscin 1 accounts for 0.1- 10%, stirred and reacted at a temperature of 75°C for 4 hours;

[0034] c. Add 6000 ml of ethanol after the reaction, filter to remove the protein precipitate, and evaporate the filtrate to dryness under reduc...

Embodiment 2

[0039] Other conditions are the same as in Example 1, except that 0.2% of the enzyme-producing inducer in the culture medium—the yam extract (dry matter) is 0.05% of flavonoids.

Embodiment 3

[0041] a. Aspergillus niger (Aspegillus niger) was cultured with agitation and ventilation at a temperature of 28-30°C for 50 days in a medium containing 5% malt extract and 1% enzyme-producing inducer—dioscin extract (dry matter). -100 hours, centrifuge and sterilize to obtain an enzyme-containing mixture, use 60-75% saturated ammonium sulfate to precipitate the enzyme protein, collect the protein, and dissolve it in acetic acid buffer (0.02M, pH5.0) of 1 / 10 of the fermentation broth volume In , dialyze to remove ammonium sulfate, and centrifuge to remove slag, which is the enzyme solution;

[0042] B. get 3 grams of dioscins, 100 milliliters of acetic acid buffer (0.02M, pH5.0) and 50 milliliters of above-mentioned enzyme liquids and mix, make 3 grams of dioscins 1 account for 0.1-10% of total reaction volume, at temperature React at 4°C for 40 hours;

[0043] c. adding 1 / 3 volume of n-butanol to extract the saponins three times, and evaporating to dryness under reduced pre...

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Abstract

The method for preparing high-pharmacodynamic low-hemolytic hypoglycosyl secondary dioscin or diosgenin by enzyme method to hydrolyze dioscin glycosyl includes the following steps: using the enzyme capable of hydrolyzing the dioscin glycosyl making the enzyme, dioscin and buffer solution be mixed and reacted for 4-40 hr., reaction temp. is 4-75 deg.C and pH value is 3-9 so as to obtain the low-glycosyl secondary dioscin or diosgenin.

Description

Technical field: [0001] The invention relates to a method for preparing low-sugar base dioscin, in particular to a method for preparing low-sugar base diosgenin by enzymatically hydrolyzing dioscin sugar base. Background technique: [0002] Dioscorea saponins are sterol saponins, which are the main active ingredients of plants of the genus Dioscorea (Dioscorea genus) in the family Dioscorea. Chinese herbal medicines of the genus Dioscorea containing dioscorea include Dioscorea nipponica Makino, Dioscorea zingiberensis G.H.Wright, Dioscorea hypoglauca, Dioscorea tocoro Makino, Dioscorea gracillima Mip, Sichuan yam (Dioscorea septemloba), hollyhock leaf yam (Dioscoreaal thacoides), etc., the rhizomes are used as medicine, and the average content of saponins is 0.93%-2.26%. [0003] The diosgenin saponins are the same, but the types and amounts of sugar groups are different, and there are many kinds of saponins. The chemical structure is (R is a sugar group): [0004] [...

Claims

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Application Information

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IPC IPC(8): C12P33/20
Inventor 金凤燮鱼红闪马白平
Owner SUNFLOWER PHARM GRP (TIANJIN) DRUG RES INST CO LTD
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