CN116200447A - A method for preparing Ginseng radix Rubri Leptoradix extract rich in ginsenoside Rg3 and Rg5 by pectase - Google Patents
A method for preparing Ginseng radix Rubri Leptoradix extract rich in ginsenoside Rg3 and Rg5 by pectase Download PDFInfo
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Abstract
The invention discloses a method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectinase, which comprises the following steps: s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:10 to 1:50, heating in water bath and stirring; s2: preparing pectase solution from pectase by using citric acid buffer solution, adding the pectase solution into the reaction system, and reacting for 24-48 h under stirring; s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 30 to 60 minutes, and then the reaction system is concentrated, dried and crushed to obtain the biotransformation dry powder of the red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5.
Description
Technical Field
The invention belongs to the technical field of medicines, and relates to a method for preparing red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5 by using pectase.
Technical Field
The natural products have unique therapeutic effects in the treatment of various diseases. Ginseng is used as a traditional Chinese herbal medicine and has prevention and treatment effects on various diseases. Ginsenoside is natural product of Ginseng radix, and has good pharmacological action. Because of the diversity of chemical structures and physicochemical properties of ginsenoside, different saponins have different pharmacological actions. The most commonly studied are Rb1, rg3, re, rd and Rh1. Among these ginsenosides, rg3 has been shown to have a wide range of pharmacological activities including anti-tumor, anti-cardiovascular disease, anti-inflammatory, anti-viral, anti-fatigue, anti-depressive and neuroprotective effects. Thus, the present study reviewed recent clinical and experimental studies on Rg3 and summarized the modern pharmacological actions of Rg3 and its mechanism of action.
Ginsenoside Rg3 is a panaxadiol tetracyclic triterpenoid saponin. Depending on the localization of hydrogen on carbon 20 (C20), ginsenoside has two stereoisomers: 20 (S) and 20 (R). Rare ginsenoside Rg3 and Rg5 are degradation Products of Protopanaxadiol (PPD), have good pharmacological activity in the aspects of protecting liver, resisting tumors, resisting hypertension, reducing blood sugar, resisting osteoarthritis, improving sleep, improving memory and the like, and become research hot spots in recent years due to low toxicity.
Rare ginsenoside 20 (S, R) -Rg3 and Rg5 can be separated from red ginseng or black ginseng, but the red ginseng or black ginseng has long participation time and low rare saponin content, and the rare ginsenoside is usually enriched and prepared from plants by adopting a traditional acid hydrolysis method, an alkaline hydrolysis method, a heating method and the like, so that the preparation method is complex, the yield is low, and the damage to human bodies, equipment and environment can be caused. There is a need for a mild, efficient and environmentally friendly method for obtaining the above ginsenoside extracts.
Disclosure of Invention
In order to overcome the technical defects, the invention adopts the following technical scheme:
a method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase comprises the following steps:
s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:10 to 1:50, heating in water bath, and stirring while heating;
s2: preparing pectase solution with concentration of 200-400U/L by using citric acid buffer solution, adding the pectase solution into the reaction system to adjust the enzyme activity of the system to 0.225-1.125U/L, and reacting for 24-48 h under stirring;
s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 30-60 min, the pectase is concentrated for 5-10 h at 50-80 ℃ to obtain red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5, and the extract is dried and crushed to obtain red ginseng rootlet extract bioconversion dry powder rich in ginsenoside Rg3 and Rg 5;
in S1, the solid mass percentage of the red ginseng rootlet extract is 35-60%;
further, in S1, the pH of the citric acid buffer solution is 3.0-4.5;
in the step S1, the water bath temperature is 35-65 ℃;
in the step S1 and the step S2, the stirring rotation speed is 100-300 r/min;
further, in S2, the pectase is a food grade commercial pectase from aspergillus niger Aspergillus niger;
further, in S3, the drying temperature is 50-80 ℃;
advantageous effects
The invention adopts pectinase from Aspergillus niger (Aspergillus niger) to convert protopanaxadiol type saponin (PPD) into Rg3 and Rg5. The preparation method disclosed by the invention is mild in condition, simple and convenient, environment-friendly, and the content of ginsenoside Rg3 and Rg5 in the obtained bioconversion red ginseng extract powder is obviously improved by more than 10 times on average, so that the preparation method has a good technical effect, is beneficial to amplifying industrial production, and has good economic benefit.
Drawings
FIG. 1 HPLC chromatogram of content analysis of S-Rg3, R-Rg3, rg5 and other monomeric ginsenoside in dry powder of Leptoradix Ginseng Rubra extract before bioconversion
FIG. 2 HPLC chromatogram of S-Rg3, R-Rg3, rg5 and other monomeric ginsenoside content analysis in Ginseng radix Rubri dry powder after bioconversion
FIG. 3 influence of pH on conversion of ginsenoside Rb1 to S-Rg3, R-Rg3 and Rg5
FIG. 4 effect of temperature on conversion of ginsenoside Rb1 to S-Rg3, R-Rg3 and Rg5
FIG. 5 effect of enzyme dosage on conversion of ginsenoside Rb1 to S-Rg3, R-Rg3 and Rg5
Detailed Description
Examples 1
Example 1
A method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase comprises the following steps: s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:25 Mixing (100 mL of citric acid buffer solution and 4g of red ginseng rootlet extract), heating in a water bath, and stirring while heating, wherein the weight percentage of solid matters in the red ginseng rootlet extract is 60%, the pH of the citric acid buffer solution is 3.5, the temperature of the water bath is 55 ℃, and the stirring speed is 200r/min; s2: preparing pectase into a high-activity pectase solution by adopting a citric acid buffer solution, adding the pectase solution into the reaction system to adjust the enzyme activity of the system to 0.225 ten thousand U/L, and reacting for 48 hours under stirring, wherein the pectase is food-grade commercial pectase from Aspergillus niger Aspergillus niger, and the stirring rotating speed is 200r/min; s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 40min, then the reaction system is moved into a reduced pressure concentration pot to be concentrated for about 6h under reduced pressure at 55 ℃ to obtain red ginseng rootlet extract extractum rich in ginsenoside Rg3 and Rg5, and then the red ginseng rootlet extract extractum is dried and crushed in an oven at 60 ℃ to obtain the red ginseng rootlet extract bioconversion dry powder rich in ginsenoside S-Rg3, R-Rg3 and Rg5.
The contents of S-Rg3, R-Rg3 and Rg5 in the red ginseng rootlet extract bioconverter powder are 17.67mg/g, 13.37mg/g and 16.37mg/g respectively.
Example 2
A method for enriching ginsenoside S-Rg3, R-Rg3 and Rg5 in red ginseng rootlet extract by using pectase comprises the following steps: s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:10 Mixing (100 mL of citric acid buffer solution and 10g of red ginseng rootlet extract), heating in a water bath, and stirring while heating, wherein the weight percentage of solid matters in the red ginseng rootlet extract is 60%, the pH of the citric acid buffer solution is 3.6, the temperature of the water bath is 55 ℃, and the stirring speed is 300r/min; s2: preparing pectase into pectase solution by using citric acid buffer solution, adding the pectase buffer solution into the reaction system to adjust the enzyme activity of the system to 1.125 ten thousand U/L, and reacting for 36h under stirring, wherein the pectase is food-grade commercial pectase from Aspergillus niger Aspergillus niger, and the stirring rotating speed is 300r/min; s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 50min, then the reaction system is moved into a reduced pressure concentration pot to be concentrated for about 6h under reduced pressure at 60 ℃ to obtain red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5, and then the red ginseng rootlet extract is dried and crushed in an oven at 60 ℃ to obtain the red ginseng rootlet extract bioconversion dry powder rich in ginsenoside Rg3 and Rg5.
The contents of S-Rg3, R-Rg3 and Rg5 in the red ginseng rootlet extract bioconverter powder are 22.70mg/g, 27.45mg/g and 17.07mg/g respectively.
Example 3
A method for enriching ginsenoside S-Rg3, R-Rg3 and Rg5 in red ginseng rootlet extract by using pectase comprises the following steps: s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:10 Mixing (100 mL of citric acid buffer solution and 10g of red ginseng rootlet extract), heating in a water bath, and stirring while heating, wherein the weight percentage of solid matters in the red ginseng rootlet extract is 60%, the pH of the citric acid buffer solution is 3.6, the temperature of the water bath is 55 ℃, and the stirring speed is 300r/min; s2: preparing pectase into pectase solution by using citric acid buffer solution, adding the pectase buffer solution into the reaction system to adjust the enzyme activity of the system to about 1.4 ten thousand U/L, and reacting for 36h under stirring, wherein the pectase is food-grade commercial pectase from Aspergillus niger Aspergillus niger, and the stirring rotating speed is 250r/min; s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 60min, then the reaction system is moved into a reduced pressure concentration pot to be concentrated for about 5h under reduced pressure at 60 ℃ to obtain red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5, and then the red ginseng rootlet extract is dried and crushed in an oven at 60 ℃ to obtain the red ginseng rootlet extract bioconversion dry powder rich in ginsenoside Rg3 and Rg5. The contents of S-Rg3, R-Rg3 and Rg5 in the red ginseng rootlet extract bioconverter powder are respectively 20.31mg/g, 26.83mg/g and 16.15mg/g.
Example 4
A method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase comprises the following steps: s1: mixing 15.310kg of red ginseng rootlet extract (solid content is about 60 percent of total extract mass percent) with 150L of citric acid buffer solution, putting the mixture into a 300L interlayer stainless steel reaction kettle, heating by adopting a water bath, and stirring while heating, wherein Rb1 content in dry powder of the red ginseng rootlet extract is 30-50 mg/g, pH of the citric acid buffer solution is 3.6, the water bath temperature is 55 ℃, and the stirring rotating speed is 200r/min; s2: preparing pectase into a pectase buffer solution by adopting a citric acid buffer solution, adding the pectase buffer solution into the reaction system to adjust the enzyme activity of the system to about 1.125 ten thousand U/L, and reacting for 48 hours under stirring, wherein the pectase is food-grade commercial pectase from Aspergillus niger Aspergillus niger, and the stirring rotating speed is 200r/min; s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 30min, then the reaction system is moved into a reduced pressure concentration pot to be concentrated for about 8h under reduced pressure at 65 ℃ to obtain about 20 kg (the solid content is 39.1 w/w%) of red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5, and then a proper amount of the extract is taken to be dried in an oven at 60 ℃ and crushed to obtain the biological conversion dry powder of red ginseng rootlet extract rich in ginsenoside S-Rg3, R-Rg3 and Rg5.
Referring to fig. 1, 2 and table 1, the contents of S-Rg3, R-Rg3 and Rg5 in the dry powder of red ginseng rootlet extract and the dry powder of bioconversion thereof were measured by HPLC analysis method, and the contents of S-Rg3, R-Rg3 and Rg5 in the dry powder of red ginseng rootlet extract and the dry powder of bioconversion thereof were 18.24mg/g, 20.35mg/g and 16.76mg/g, respectively.
TABLE 1 comparison of the content of ginsenoside monomers in Ginseng radix Rubri extract and Ginseng radix Rubri extract bioconversion Dry powder
Experimental example 2
2.1 screening of experimental conditions
As shown in FIGS. 3-5, the reaction conditions in actual industrial production were finally determined according to the changes in the amounts of S-Rg3, R-Rg3 and Rg5 produced by 1.5mg of ginsenoside Rb1 monomer in 6mL of citric acid buffer solutions with different pH values, different temperatures and different enzyme dosage ranges.
(1) As shown in FIG. 3, the production of S-Rg3, R-Rg3 and Rg5 was highest at pH3.6 in the optimum pH range of the pectinase;
(2) As shown in FIG. 4, the formation of S-Rg3, R-Rg3 and Rg5 was high at temperatures of 55℃and 60℃in the optimum temperature range of pectinase, and similar results were exhibited. From the viewpoint of production cost, the optimal temperature is preferably 60 ℃;
(3) As shown in FIG. 5, the amounts of pectinase used were 3.750U and 7.500U, the amounts of S-Rg3, R-Rg3 and Rg5 produced were higher than those obtained when 1.875U was used, and both groups exhibited similar results. From the viewpoint of production cost, the enzyme amount of 3.750U is preferable.
The foregoing is merely a preferred embodiment of the present invention, and it should be noted that modifications and variations could be made by those skilled in the art without departing from the technical principles of the present invention, and such modifications and variations should also be regarded as being within the scope of the invention.
Claims (7)
1. A method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by using pectase is characterized by comprising the following steps:
s1: the red ginseng rootlet extract and the citric acid buffer solution are mixed according to the mass ratio of 1:10 to 1:50, heating in water bath, and stirring while heating;
s2: preparing pectase solution with concentration of 200-400U/L by using citric acid buffer solution, adding the pectase solution into the reaction system to adjust the enzyme activity of the system to 0.225-1.125U/L, and reacting for 24-48 h under stirring;
s3: after the reaction is finished, the temperature of the reaction system is increased to more than 80 ℃ to inactivate pectase for 30-60 min, the pectase is concentrated for 5-10 h at 50-80 ℃ to obtain red ginseng rootlet extract rich in ginsenoside Rg3 and Rg5, and the extract is dried and crushed to obtain the red ginseng rootlet extract bioconversion dry powder rich in ginsenoside Rg3 and Rg5.
2. The method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase according to claim 1, wherein in S1, the solid mass percentage of the red ginseng rootlet extract is 35-60%.
3. The method for enriching ginsenoside Rg3 and Rg5 in Leptoradix Ginseng Rubra extract with pectase according to claim 1, wherein in S1, the pH of the citric acid buffer solution is 3.0-4.5.
4. The method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase according to claim 1, wherein in S1, the water bath temperature is 35-65 ℃.
5. The method for enriching ginsenoside Rg3 and Rg5 in Leptoradix Ginseng Rubra extract with pectase according to claim 1, wherein in S1 and S2, the stirring speed is 100-300 r/min.
6. The method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase according to claim 1, wherein in S2, the pectase is food-grade commercial pectase from Aspergillus niger Aspergillus niger.
7. The method for enriching ginsenoside Rg3 and Rg5 in red ginseng rootlet extract by pectase according to claim 1, wherein in S3, the drying temperature is 50-80 ℃.
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| CN117883347A (en) * | 2024-03-13 | 2024-04-16 | 广州梵之容化妆品有限公司 | Extraction and conversion method for preparing rare ginsenoside and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117883347A (en) * | 2024-03-13 | 2024-04-16 | 广州梵之容化妆品有限公司 | Extraction and conversion method for preparing rare ginsenoside and application thereof |
| CN117883347B (en) * | 2024-03-13 | 2024-05-28 | 广州梵之容化妆品有限公司 | Extraction and conversion method for preparing rare ginsenoside and application thereof |
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