CN102477455A - Method for preparing mogroside IV - Google Patents
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- CN102477455A CN102477455A CN2010105610299A CN201010561029A CN102477455A CN 102477455 A CN102477455 A CN 102477455A CN 2010105610299 A CN2010105610299 A CN 2010105610299A CN 201010561029 A CN201010561029 A CN 201010561029A CN 102477455 A CN102477455 A CN 102477455A
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Abstract
The invention which relates to a method for preparing mogroside IV belongs to the traditional Chinese medicinal field. A technical problem to be solved in the invention is to provide a low cost method for preparing the mogroside IV. The method for preparing the mogroside IV in the invention is characterized in that mogroside V, Momordica grosvenori, or a Momordica grosvenori extract containing the mogroside V is hydrolyzed with beta-glucosidase. The method of the invention, which has the advantages of simple technology and low cost, allows the content of the mogroside IV in Momordica grosvenori powder to be improved to above 1.5% from below 0.05%; and the method allows the content of the mogroside IV in the Momordica grosvenori extract to be improved to above 20%, wherein the content of the mogroside V in the Momordica grosvenori extract is 40%, and the original content of the mogroside IV in the Momordica grosvenori extract is 1.5%. The purity of the mogroside IV can reach above 98% after separation and purification. The method of the invention, which provides a new approach for the preparation of the mogroside IV, has a wide application prospect.
Description
Technical field
The present invention relates to prepare the method for Momordia grosvenori aglycone IV, belong to the Chinese drug field.
Background technology
The Grosvenor Momordica main product is in Guangxi, Jiangxi, and Guangdong also is known as the monk fruit, has the plantation in hundreds of years historical in China, and its application in the traditional Chinese medical science can be traced back to 1813 years.Cucurbitane Triterpene Glycosides compounds in the Grosvenor Momordica comprises Simon glycosides I, Momordia grosvenori aglycone II, Momordia grosvenori aglycone III, Momordia grosvenori aglycone IIIE, Momordia grosvenori aglycone IV, momordica grosvenori glycoside V etc.Wherein the content of Momordia grosvenori aglycone IV is extremely low, and it is high to extract the big cost of difficulty, the relevant report of not seeing the preparation technology that Momordia grosvenori aglycone IV is arranged both at home and abroad.
At present, external medical worker finds that Momordia grosvenori aglycone IV has the effect of good curing third liver, can be separately or with the shared good therapeutic action that plays of other drug.Its toxic side effect of Momordia grosvenori aglycone IV is little, and bioavailability is high, is emerging antiviral natural plant constituent, has vast market and is worth.
The bottleneck problem of restriction Momordia grosvenori aglycone IV application at present is the preparation of Momordia grosvenori aglycone IV, and Momordia grosvenori aglycone IV is content＜0.05% (content of momordica grosvenori glycoside V is about 2%) in the Grosvenor Momordica raw material powder, so glycosides IV preparation difficulty, cost is high.Serious restriction the Application and Development of Momordia grosvenori aglycone IV.Therefore, how to develop a kind of method for preparing Momordia grosvenori aglycone IV efficiently and become the technical barrier that this area presses for solution at present.
Summary of the invention
Technical problem to be solved by this invention provides a kind of lower-cost method for preparing Momordia grosvenori aglycone IV.
The method that the present invention prepares Momordia grosvenori aglycone IV is: adopt beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V, Grosvenor Momordica (can be Grosvenor Momordica fresh fruit or Grosvenor Momordica dry powder) or contain the Fructus Monordicae extract of momordica grosvenori glycoside V.
Wherein, Grosvenor Momordica water extract or ethanolic soln extract all are applicable to the present invention.
Further, the above-mentioned method for preparing Momordia grosvenori aglycone IV, preferred every kg momordica grosvenori glycoside V adopts 5k～50k ten thousand U beta-glucoside enzymic hydrolysiss; Wherein, 1U be beta-glucosidase under the condition of optimum pH and temperature, the beta-glucan solution of 1min hydrolysis 1wt% produces the consumption of the enzyme of 1 μ g glucose.The hydrolysis time of beta-glucosidase is preferably 2～24h.
Wherein, the above-mentioned method for preparing Momordia grosvenori aglycone IV, it adopts beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V or contains the concrete steps of Fructus Monordicae extract of momordica grosvenori glycoside V following:
A, dissolving: momordica grosvenori glycoside V, Grosvenor Momordica or contain the Fructus Monordicae extract solubilizing agent dissolving of momordica grosvenori glycoside V, the concentration that makes the momordica grosvenori glycoside V in the solution is 0.025～0.300g/mL;
B, hydrolysis: add the beta-glucoside enzymic hydrolysis in a step gained solution, and the pH value of regulator solution and temperature adapt the enzymolysis pH value and the temperature of itself and beta-glucosidase;
C, separation, purifying obtain Momordia grosvenori aglycone IV.
Wherein, the solvent in the said a step can be conventional solvent, the damping fluid that solvent is preferably water or adapts with the enzymolysis pH value of beta-glucosidase.
Wherein, during above-mentioned a step enzymolysis,, reduce enzymic hydrolysis efficient if the momordica grosvenori glycoside V excessive concentration can form intermolecular ydrogen bonding in solution; If momordica grosvenori glycoside V concentration is low excessively, then the productive rate of Momordia grosvenori aglycone IV is lower, also can increase concentrated cost, and the concentration of the momordica grosvenori glycoside V in the solution is that 0.025～0.300g/mL is comparatively suitable.
Further,, add too much, then can cause waste because the solubleness of beta-glucosidase is limited; The concentration of beta-glucosidase is low excessively, and then hydrolysis efficiency is lower, and the concentration of the beta-glucosidase during above-mentioned b step hydrolysis in the solution is preferably 1.5 ten thousand～5.0 ten thousand U/mL (starting point concentration of enzyme when referring to hydrolysis).
Wherein, preferred hydrolysising condition is: the momordica grosvenori glycoside V concentration 0.1g/mL during hydrolysis in the solution (starting point concentration of momordica grosvenori glycoside V when referring to hydrolysis), and the beta-glucoside enzyme concn is 2.0 ten thousand U/mL (starting point concentrations of enzyme when referring to hydrolysis), hydrolysis time is 16h.
Wherein, Because the kind of beta-glucosidase is more; The pH value that the best enzyme of different beta-glucosidases is lived is different with temperature, therefore, and in order to improve hydrolysis efficiency; PH value and the temperature that can regulate hydrating solution adapts the enzymolysis pH value and the temperature of itself and beta-glucosidase, promptly is adjusted to pH value and temperature that the best enzyme of beta-glucosidase is lived.
Wherein, can adopt conventional separation, purification process in the above-mentioned c step, as: gel filtration chromatography method, HPLC method of purification etc., separation, purifying obtain highly purified Momordia grosvenori aglycone IV product.
Wherein, Above-mentioned beta-glucosidase can be the beta-glucosidase of routine; The beta-glucosidase (beta-glucosidase can prepare by ordinary method) that can adopt commercially available beta-glucosidase or prepare voluntarily; Consider the factor of cost, above-mentioned beta-glucosidase is preferably cellulase, 1, and is at least a in 4-callose glucoside lytic enzyme, the hemicellulase.
The inventive method technology is simple, and cost is lower, can bring up to more than 2.00% with the content of Momordia grosvenori aglycone IV in the Grosvenor Momordica Fruit from＜0.05%; For Fructus Monordicae extract (momordica grosvenori glycoside V content is about 40%, and Momordia grosvenori aglycone IV content is about 1.5%), the inventive method can be brought up to the content of Momordia grosvenori aglycone IV more than 20%.Through separation, purifying, the purity of Momordia grosvenori aglycone IV is reached more than 98%.The inventive method is that the preparation of Momordia grosvenori aglycone IV provides a kind of new approach, has broad application prospects.
Description of drawings
Fig. 1 is the HPLC figure of Fructus Monordicae extract before and after the enzymic hydrolysis, and wherein, figure A is the HPLC figure of the Fructus Monordicae extract before the enzymic hydrolysis, and figure B is the HPLC figure of Fructus Monordicae extract after the enzymic hydrolysis, and drawing the circle place among the figure is Momordia grosvenori aglycone IV.
Fig. 2 is the HPLC figure (figure A) of the Momordia grosvenori aglycone IV after purified and the HPLC figure (figure B) of Momordia grosvenori aglycone IV standard model.
Fig. 3 be the Momordia grosvenori aglycone IV behind the purifying nuclear magnetic spectrum (
1H NMR).
Fig. 4 be the Momordia grosvenori aglycone IV behind the purifying nuclear magnetic spectrum (
13C NMR).
Fig. 5 is the mass spectrum of the Momordia grosvenori aglycone IV behind the purifying.
Embodiment
The structure of momordica grosvenori glycoside V is shown in (I), and the structure of Momordia grosvenori aglycone IV is shown in (II).
Can find out from structural formula (I) with (II): Momordia grosvenori aglycone IV lacks a glucose than momordica grosvenori glycoside V, in theory, can the glycosyl of momordica grosvenori glycoside V reduced with glycosyl hydrolase, acid or basic hydrolysis, thereby obtain Momordia grosvenori aglycone IV.But, find through a large amount of tests, adopt acid or basic hydrolysis momordica grosvenori glycoside V; Its targeted is not strong; Destroy easily the Momordia grosvenori aglycone structure, can not get Momordia grosvenori aglycone IV or the Momordia grosvenori aglycone IV productive rate that obtains lower, be difficult to satisfy the needs of industrial mass production Momordia grosvenori aglycone IV.Contriver of the present invention is through a large amount of tests; Find the beta-glucoside enzyme dosage that adjustment is suitable, and control suitable enzymolysis time, but can be with the content of the glycosides IV of cucurbitane Triterpene Glycosides compounds in the Grosvenor Momordica from＜0.05%; Bring up to more than 2.00%; For Fructus Monordicae extract (momordica grosvenori glycoside V content is about 40%, and Momordia grosvenori aglycone IV content is about 1.50%), the inventive method can be brought up to the content of Momordia grosvenori aglycone IV more than 20%.All significantly exceeded the Momordia grosvenori aglycone IV that estimates productive rate＜0.05% (is raw material with the mangosteen powder) and＜1.50% (is raw material with the arhat extract); Productive rate＜0.11% (is raw material with the arhat extract) of farther Momordia grosvenori aglycone IV far above acid or basic hydrolysis and＜2.20% (is raw material with the arhat extract), reached the requirement of industrial mass production Momordia grosvenori aglycone IV.
The method that the present invention prepares Momordia grosvenori aglycone IV is: adopt beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V, Grosvenor Momordica or contain the Fructus Monordicae extract of momordica grosvenori glycoside V.Wherein, Grosvenor Momordica water extract or ethanolic soln extract all are applicable to the present invention.
Further, in order to improve the productive rate of Momordia grosvenori aglycone IV, the above-mentioned method for preparing Momordia grosvenori aglycone IV, preferred every kg momordica grosvenori glycoside V employing 5k～50k ten thousand U beta-glucoside enzymic hydrolysiss. Wherein, 1U be beta-glucosidase under the condition of optimum pH and temperature, the beta-glucan solution of 1min hydrolysis 1wt% produces the consumption of the enzyme of 1 μ g glucose.The hydrolysis time of beta-glucosidase is preferably 2～24h.
Wherein, the above-mentioned method for preparing Momordia grosvenori aglycone IV, it adopts beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V or contains the concrete steps of Fructus Monordicae extract or Grosvenor Momordica raw material itself of momordica grosvenori glycoside V following:
A, dissolving: momordica grosvenori glycoside V, Grosvenor Momordica or contain the Fructus Monordicae extract solubilizing agent dissolving of momordica grosvenori glycoside V, the concentration that makes the momordica grosvenori glycoside V in the solution is 0.025～0.300g/mL;
B, hydrolysis: add the beta-glucoside enzymic hydrolysis in a step gained solution, and the pH value of regulator solution and temperature adapt the enzymolysis pH value and the temperature of itself and beta-glucosidase;
C, separation, purifying obtain Momordia grosvenori aglycone IV.
Wherein, the solvent in the said a step can be conventional solvent, the damping fluid that solvent is preferably water or adapts with the enzymolysis pH value of beta-glucosidase.
Wherein, during above-mentioned a step enzymolysis,, reduce enzymic hydrolysis efficient if the momordica grosvenori glycoside V excessive concentration can form intermolecular ydrogen bonding in solution; If momordica grosvenori glycoside V concentration is low excessively, then the productive rate of Momordia grosvenori aglycone IV is lower, also can increase concentrated cost, and the concentration of the momordica grosvenori glycoside V in the solution is that 0.025～0.300g/mL is comparatively suitable.
Further,, add too much, then can cause waste because the solubleness of beta-glucosidase is limited; The concentration of beta-glucosidase is low excessively, and then hydrolysis efficiency is lower, and the concentration of the beta-glucosidase during above-mentioned b step hydrolysis in the solution is preferably 1.5 ten thousand～5.0 ten thousand U/mL.
Wherein, preferred hydrolysising condition is: the momordica grosvenori glycoside V concentration 0.1g/mL during hydrolysis in the solution (starting point concentration of momordica grosvenori glycoside V when referring to hydrolysis), and the beta-glucoside enzyme concn is 2.0 ten thousand U/mL (starting point concentrations of enzyme when referring to hydrolysis), hydrolysis time is 16h.
Wherein, Because the kind of beta-glucosidase is more; The pH value that the best enzyme of different beta-glucosidases is lived maybe be different with temperature, therefore, and in order to improve hydrolysis efficiency; PH value and the temperature that can regulate hydrating solution adapts the enzymolysis pH value and the temperature of itself and beta-glucosidase, promptly is adjusted to pH value and temperature that the best enzyme of beta-glucosidase is lived.
Wherein, can adopt conventional separation, purification process in the above-mentioned c step, as: gel filtration chromatography method, HPLC method of purification etc., separation, purifying obtain the highly purified Momordia grosvenori aglycone IV product that obtains.
Wherein, Above-mentioned beta-glucosidase can be the beta-glucosidase of routine; The beta-glucosidase (beta-glucosidase can prepare by ordinary method) that can adopt commercially available beta-glucosidase or prepare voluntarily; Consider the factor of cost, above-mentioned beta-glucosidase is preferably cellulase, 1, and is at least a in 4-callose glucoside lytic enzyme, the hemicellulase.
Do further description below in conjunction with embodiment and Test Example specific embodiments of the invention, therefore do not limit the present invention among the described scope of embodiments.
The shaker test of Test Example 1 beta-glucoside enzyme dosage
Raw material is about 40.00% (content of momordica grosvenori glycoside V among the present invention or Momordia grosvenori aglycone IV all refers to mass content) of momordica grosvenori glycoside V content, Momordia grosvenori aglycone IV content＜1.00%.The beta-glucan enzymic hydrolysis to jade of the He family Bioisystech Co., Ltd is purchased in employing, and the enzyme concn during hydrolysis (referring to starting point concentration) is respectively 0.5 ten thousand U/mL, 2.5 ten thousand U/mL, 5.0 ten thousand U/mL, hydrolysis 12h, the yield of mensuration Momordia grosvenori aglycone IV.The result is as shown in table 1.Can find out that from table 1 when enzyme concn was 2.5 ten thousand～5.0 ten thousand U/mL, Momordia grosvenori aglycone IV yield was higher.
Table 1
The shaker test of Test Example 2 hydrolysis times
Raw material is that momordica grosvenori glycoside V content is about 40.00%, Momordia grosvenori aglycone IV content＜1.00%.The beta-glucan enzymic hydrolysis to jade of the He family Bioisystech Co., Ltd is purchased in employing; Enzyme concn during hydrolysis (referring to starting point concentration) is 2.5 ten thousand U/mL; Momordica grosvenori glycoside V concentration is 0.05g/mL, hydrolysis 2,8,16,24,48 respectively, 72h, the yield of mensuration Momordia grosvenori aglycone IV.The result sees table 2.Can find out that from table 2 when hydrolysis time was 8～24h, the yield of Momordia grosvenori aglycone IV was higher.
Table 2
The shaker test of momordica grosvenori glycoside V concentration during Test Example 3 hydrolysis
Raw material is about 40.00% Fructus Monordicae extract of momordica grosvenori glycoside V content, wherein Momordia grosvenori aglycone IV content＜1.00%.The beta-glucan enzymic hydrolysis to jade of the He family Bioisystech Co., Ltd is purchased in employing; Enzyme concn during hydrolysis (referring to starting point concentration) is 2.5 ten thousand U/mL; Momordica grosvenori glycoside V concentration is respectively 0.05g/mL, 0.25g/mL, 0.50g/mL, hydrolysis 16h, the yield of mensuration Momordia grosvenori aglycone IV.The result sees table 3.Can find out that from table 3 when momordica grosvenori glycoside V concentration was 0.05～0.50g/mL during hydrolysis, the yield of Momordia grosvenori aglycone IV was all higher, wherein, when momordica grosvenori glycoside V concentration was 0.25g/mL, the yield of Momordia grosvenori aglycone IV was the highest.
Table 3
Test Example 4 orthogonal tests
Raw material is about 40% Fructus Monordicae extract of momordica grosvenori glycoside V content, wherein Momordia grosvenori aglycone IV content＜1.00%.Result according to Test Example 1～3 determines experimental factor and level, and the design orthogonal table, measures the top condition of hydrolysis.The result sees table 4～6.
Table 4 experimental factor and level
Table 5 orthogonal table and test-results
Table 6 test result analysis
Can find out that from table 4～5 through analyzing, the best of breed that theory obtains is: momordica grosvenori glycoside V concentration 0.10g/mL, enzyme concn 3.5 ten thousand U/mL, hydrolysis time 16h.
Can find out that from table 4～6 during hydrolysis, momordica grosvenori glycoside V concentration shows as utmost point significant difference; Because of excessive concentration forms hydrogen bond between the molecule in Momordia grosvenori aglycone solution; The hydrolysis of the enzyme that hinders not is that concentration is high more good more so show, but best value is arranged in certain scope.The influence of hydrolysis time is after momordica grosvenori glycoside V concentration, before enzyme concn.Time is not that the longer the better yet, because glycosides IV also can be continued to be hydrolyzed conversion.Wherein enzyme concn influences inapparent reason and infers and determined its state that reached capacity for the solubleness of enzyme concn in solution.Because the influence of enzyme is not remarkable relatively, take all factors into consideration the cost reason again, enzyme concn selects 1.5 to 2.5 ten thousand U/mL preferable.
In sum, optimum hydrolysising condition is preferably momordica grosvenori glycoside V concentration 0.10g/mL, enzyme concn 2.0 ten thousand U/mL, hydrolysis time 16h.Reappearing test-results is that Momordia grosvenori aglycone IV content is 21.78%.
With dropping into after the fresh Fructus Momordicae 1000kg fragmentation in the extractor, to extract 3 times down in slight boiling condition with the 4000kg purified water, extraction time is 1.5h, 1h, 1h, the filter residue after each the filtration gets into next time and extracts, and merging filtrate is concentrated into 4000L.Recording momordica grosvenori glycoside V content is 0.27% of Grosvenor Momordica fresh weight, and Momordia grosvenori aglycone IV content is less than 0.01%.Temperature is reduced to 50 ℃ and is added 10g beta-glucanase (jade of the He family Bioisystech Co., Ltd produces, down together), transfers pH to 5, hydrolysis time 20h.Recording momordica grosvenori glycoside V content is 0.10%, Momordia grosvenori aglycone IV content 0.12%.Suction filtration, filtrating is concentrated into certain volume, adds 10%Ca (OH)
2Precipitated impurities, quiet system is filtered; Supernatant is with extracted with diethyl ether ester solubility impurity; Discard the ether part, water continues to adopt D280 polymeric adsorbent (Haiyang Chemical Plant, Qingdao) and the refining carbohydrate, protein-based and pigment molecular removed of 100-200 order silica gel (Haiyang Chemical Plant, Qingdao); Collect the Momordia grosvenori aglycone part, transfer pH to 5 with 1mol/L hydrochloric acid.Adopt the decolouring of 201 type strong basic ion exchange resins.Be concentrated into suitable volumes, through preparation HPLC purifying repeatedly, concentrate, drying obtains sample.HPLC detects and obtains Momordia grosvenori aglycone IV0.97kg, and its content is more than 98%.The HPLC figure of Momordia grosvenori aglycone IV after purified and the HPLC figure of Momordia grosvenori aglycone IV standard model are as shown in Figure 2.The nuclear magnetic spectrum of Momordia grosvenori aglycone IV behind the purifying is as shown in Figure 3, and the mass spectrum of the Momordia grosvenori aglycone IV behind the purifying is as shown in Figure 4.
Embodiment 2 adopts the inventive method to prepare Momordia grosvenori aglycone IV
Grosvenor Momordica dry powder 10kg is dropped in the extractor, and with 50L 50% alcohol reflux 3 times, extraction time is 2h at every turn; Filter residue after each the filtration gets into next time and extracts; It is 8.4% that concentrate drying medicinal extract 1kg, HPLC record momordica grosvenori glycoside V content, and Momordia grosvenori aglycone IV content is less than 0.1%.Medicinal extract redissolves in 5L water, the elimination insolubles, and temperature rises to 50 ℃ and adds 10g1, and 4-callose glucoside lytic enzyme is transferred pH to 4.8, hydrolysis time 20h, recording momordica grosvenori glycoside V content is 3.4%, Momordia grosvenori aglycone IV content is 4.2%.Adopt embodiment 1 scheme purifying, obtain Momordia grosvenori aglycone IV0.17kg, its content is more than 98%.
40% momordica grosvenori glycoside V raw material 1kg is dropped in the extractor, use the 5L water dissolution, temperature rises to 50 ℃ and adds the 10g beta-glucanase, transfers pH to 5, and hydrolysis time 16h records momordica grosvenori glycoside V content 15.5%, Momordia grosvenori aglycone IV content 17.3%.Concentrate, through preparation HPLC purifying repeatedly, concentrate, drying obtains Momordia grosvenori aglycone IV0.157kg, and its content is more than 98%.The HPLC figure of Momordia grosvenori aglycone IV content is as shown in Figure 1 in the Fructus Monordicae extract of hydrolysis front and back, and as can beappreciated from fig. 1, the Momordia grosvenori aglycone IV content after the hydrolysis is obviously higher.
With embodiment 1, only wherein beta-glucanase (jade of the He family Bioisystech Co., Ltd) changes cellulase (jade of the He family Bioisystech Co., Ltd) into, obtains Momordia grosvenori aglycone IV1.02kg at last, and its content is more than 98%.
With embodiment 2, only wherein beta-glucosidase (jade of the He family Bioisystech Co., Ltd beta-glucosidase) changes hemicellulase into, obtains Momordia grosvenori aglycone IV0.16kg at last, and its content is more than 98%.
Claims (9)
1. prepare the method for Momordia grosvenori aglycone IV, it is characterized in that: adopt beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V, Grosvenor Momordica or contain the Fructus Monordicae extract of momordica grosvenori glycoside V.
2. the method for preparing Momordia grosvenori aglycone IV according to claim 1 is characterized in that: described Fructus Monordicae extract is Grosvenor Momordica water extract or ethanolic soln extract.
3. the method for preparing Momordia grosvenori aglycone IV according to claim 1 and 2 is characterized in that: every g momordica grosvenori glycoside V adopts the beta-glucoside enzymic hydrolysis of 5～500,000 U; Wherein, 1U be beta-glucosidase under the condition of optimum pH and temperature, the beta-glucan solution of 1min hydrolysis 1wt% produces the consumption of the enzyme of 1 μ g glucose.
4. according to each described method for preparing Momordia grosvenori aglycone IV of claim 1～3, it is characterized in that: the hydrolysis time of beta-glucosidase is 2～24h.
5. according to each described method for preparing Momordia grosvenori aglycone IV of claim 1～4, it is characterized in that: adopt beta-glucoside enzymic hydrolysis momordica grosvenori glycoside V or contain the concrete steps of Fructus Monordicae extract of momordica grosvenori glycoside V following:
A, dissolving: momordica grosvenori glycoside V, Grosvenor Momordica or contain the Fructus Monordicae extract solubilizing agent dissolving of momordica grosvenori glycoside V, the concentration that makes the momordica grosvenori glycoside V in the solution is 0.025～0.300g/mL;
B, hydrolysis: add the beta-glucoside enzymic hydrolysis in a step gained solution, and the pH value of regulator solution and temperature adapt the enzymolysis pH value and the temperature of itself and beta-glucosidase;
C, separation, purifying obtain Momordia grosvenori aglycone IV.
6. the method for preparing Momordia grosvenori aglycone IV according to claim 5 is characterized in that: the solvent in the said a step is water or the damping fluid that adapts with the enzymolysis pH value of beta-glucosidase.
7. according to claim 5 or the 6 described methods that prepare Momordia grosvenori aglycone IV, it is characterized in that: the concentration of the beta-glucosidase during said b step hydrolysis in the solution is 1.5 ten thousand～5.0 ten thousand U/mL.
8. the method for preparing Momordia grosvenori aglycone IV according to claim 5 is characterized in that: the momordica grosvenori glycoside V concentration 0.1g/mL during hydrolysis in the solution, and the beta-glucoside enzyme concn is 2.0 ten thousand U/mL, hydrolysis time is 16h.
9. according to each described method for preparing Momordia grosvenori aglycone IV of claim 1～8, it is characterized in that: described beta-glucosidase is a cellulase, 1, and is at least a in 4-callose glucoside lytic enzyme, the hemicellulase.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010561029.9A CN102477455B (en) | 2010-11-26 | 2010-11-26 | Method for preparing mogroside IV |
| PCT/CN2011/073861 WO2012068832A1 (en) | 2010-11-26 | 2011-05-10 | Method for preparing mogroside iv |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201010561029.9A CN102477455B (en) | 2010-11-26 | 2010-11-26 | Method for preparing mogroside IV |
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| CN102477455B CN102477455B (en) | 2014-03-26 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107281463A (en) * | 2017-08-23 | 2017-10-24 | 上海新肌生物科技有限公司 | A kind of comprehensive radicals scavenging method for the free radical generating source that can remove 99% |
| CN109055474A (en) * | 2018-10-31 | 2018-12-21 | 大闽食品（漳州）有限公司 | A method of improving mogroside sugariness |
| JPWO2018016483A1 (en) * | 2016-07-19 | 2019-06-20 | サントリーホールディングス株式会社 | Method of producing mogrol or mogrol glycoside |
| CN114592028A (en) * | 2020-12-07 | 2022-06-07 | 成都普睿法药物研发有限公司 | Method for preparing notoginsenoside Fd and esculin XIII |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016117549A1 (en) * | 2015-01-20 | 2016-07-28 | サントリーホールディングス株式会社 | Method for preparing mogroside |
| US11180789B2 (en) * | 2019-09-26 | 2021-11-23 | National Taiwan University | Method for bioconversion of mogroside extracts into siamenoside I |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2003277270A (en) * | 2002-03-20 | 2003-10-02 | Univ Nihon | Carcinogensis preventing agent |
| CN101200753A (en) * | 2007-08-09 | 2008-06-18 | 桂林惠通生物科技有限公司 | Fructus momordicae extract with mogroside V content being more than or equal to 40% and preparation method thereof |
| JP4147038B2 (en) * | 2002-03-01 | 2008-09-10 | サラヤ株式会社 | Rakan fruit glycosides with improved taste and method for producing the same |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101664138B (en) * | 2009-09-27 | 2012-05-09 | 何伟平 | Fructus momordicae instant powder and preparation method thereof |
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2010
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP4147038B2 (en) * | 2002-03-01 | 2008-09-10 | サラヤ株式会社 | Rakan fruit glycosides with improved taste and method for producing the same |
| JP2003277270A (en) * | 2002-03-20 | 2003-10-02 | Univ Nihon | Carcinogensis preventing agent |
| CN101200753A (en) * | 2007-08-09 | 2008-06-18 | 桂林惠通生物科技有限公司 | Fructus momordicae extract with mogroside V content being more than or equal to 40% and preparation method thereof |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPWO2018016483A1 (en) * | 2016-07-19 | 2019-06-20 | サントリーホールディングス株式会社 | Method of producing mogrol or mogrol glycoside |
| EP3502265A4 (en) * | 2016-07-19 | 2020-03-25 | Suntory Holdings Limited | Method for producing mogrol or mogrol glycoside |
| US11008600B2 (en) | 2016-07-19 | 2021-05-18 | Suntory Holdings Limited | Method for producing mogrol or mogrol glycoside |
| CN107281463A (en) * | 2017-08-23 | 2017-10-24 | 上海新肌生物科技有限公司 | A kind of comprehensive radicals scavenging method for the free radical generating source that can remove 99% |
| CN109055474A (en) * | 2018-10-31 | 2018-12-21 | 大闽食品（漳州）有限公司 | A method of improving mogroside sugariness |
| CN114592028A (en) * | 2020-12-07 | 2022-06-07 | 成都普睿法药物研发有限公司 | Method for preparing notoginsenoside Fd and esculin XIII |
| CN114592028B (en) * | 2020-12-07 | 2024-02-09 | 成都普睿法药物研发有限公司 | Method for preparing notoginsenoside Fd and esculin XIII |
Also Published As
| Publication number | Publication date |
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| CN102477455B (en) | 2014-03-26 |
| WO2012068832A1 (en) | 2012-05-31 |
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