Probiotic fermentation process for improving content of ginsenoside
By inoculating compound probiotics in American ginseng solution to extract ginseng saponins, the problems of low efficiency of traditional extraction methods and easy destruction of active ingredients are solved, the content and purity of ginseng saponins are improved, and its effect in cosmetics is enhanced.
Patent Information
- Application Number
- CN202411738607.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-29
- Publication Date
- 2025-02-28
AI Technical Summary
The traditional ginseng saponin extraction method has limited efficiency, which may destroy the active ingredients, resulting in limited purity and extraction rate, and will not be able to perform better results in cosmetics.
Complex probiotics are used to inoculate ferment and extract ginseng saponin from American ginseng solution. The American ginseng solution prepared through specific processes and components provides a suitable nutritional environment and improves the conversion rate and stability of ginseng saponin.
It improves the content and purity of ginseng saponins, enhances its effect in cosmetics, and promotes the extraction and conversion of ginseng saponins through the metabolic activities of probiotics.
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of probiotic fermentation and relates to a probiotic fermentation process for increasing the content of ginsenosides. Background Art
[0002] As a traditional precious medicinal material, American ginseng has a long history of use. One of its main ingredients is ginsenosides, which have important pharmacological effects such as enhancing immunity and anti-fatigue. It also has antioxidant, anti-ultraviolet and skin care effects, and is widely used in the field of cosmetics. Traditional ginsenoside extraction methods such as solvent extraction and ultrasonic extraction have limited extraction efficiency, and the active ingredients may be destroyed during the extraction process, which limits the purity and extraction rate of ginsenosides and makes it impossible to achieve better effects in cosmetics. Based on this, converting the extraction method to extract ginsenosides has important practical significance, and probiotic fermentation technology, as an emerging biotechnology, has become the main research direction at present. Summary of the invention
[0003] The purpose of the present invention is to provide a probiotic fermentation process for increasing the content of ginsenosides. The present invention adopts a composite probiotic inoculated into American ginseng liquid to ferment and extract ginsenosides. The American ginseng liquid prepared by a specific process and components is rich in a variety of beneficial ingredients, which provides a suitable nutritional environment for the growth of the strain, improves the conversion rate and stability of ginsenosides, and thus increases the content of ginsenosides.
[0004] The purpose of the present invention can be achieved through the following technical solutions:
[0005] A probiotic fermentation process for increasing the content of ginsenosides, the probiotic fermentation process comprising:
[0006] The fermentation liquid of the strain is inoculated into the American ginseng liquid at an inoculation rate of 10-12wt%, the pH is adjusted to 6.2-6.8, the liquid is cultured at 30-40°C for 48-60h, and the American ginseng fermentation liquid is obtained after sterilization at 121°C for 15min.
[0007] Furthermore, the concentration of the strain in the strain fermentation broth is 1-1.2×10 7 CFU / g, and the strain is composed of bifidobacterium and lactobacillus in a ratio of 1.7-2.1:0.8-1.2; the fermentation condition of the strain fermentation liquid is anaerobic conditions, cultured at 34-38°C for 40-50h, and the fermentation is completed to obtain the strain fermentation liquid.
[0008] Furthermore, the pH of the fermentation medium used for the strain fermentation broth is 6.2-6.8, and the raw materials of the fermentation medium include 4.7-6.5 g / L natto extract, 100-120 g / L glucose, 5-6 g / L yeast extract, 1-2 g / L potassium dihydrogen phosphate, 0.3-0.7 g / L magnesium sulfate, 0.8-1 g / L ferrous sulfate and the balance deionized water.
[0009] Furthermore, the raw materials of the American ginseng liquid include, by mass percentage, 81-88wt% of American ginseng extract, 5-9wt% of natto bacteria extract, 1-2wt% of vitamin C and the balance of deionized water.
[0010] Furthermore, the preparation method of the American ginseng extract is:
[0011] Wash and dry the American ginseng, crush it through a 200-300 mesh sieve, add 10-16 times the weight of deionized water, treat it with high-pressure microfluidization twice, reflux it at 78-82°C for 1.2-2h to obtain an extract, and concentrate it by rotary evaporation to obtain the American ginseng extract.
[0012] Furthermore, the pressure and flow rate of the high-pressure microfluidic treatment are 80-120 MPa and 60-100 mL / min, respectively.
[0013] Furthermore, the rotary evaporation concentration refers to rotary evaporation at 40° C. to reduce the volume of the original extract to 1 / 4.
[0014] Furthermore, the preparation method of the Bacillus natto extract is:
[0015] The soybeans were mixed with deionized water at a solid-liquid ratio of 1:2-6 g / mL, soaked for 3-5 hours, drained and steamed in a 100°C steamer for 20-30 minutes, taken out and cooled, mixed with deionized water at a solid-liquid ratio of 1:4-10 g / mL, heated to 37-41°C, stirred at 50-100 rmp for 1-2 hours to obtain an extract, added with a supplement, adjusted to pH 6.5-7.5, continued stirring to obtain soybean liquid, inoculated, fermented, centrifuged and obtained a supernatant to obtain a Bacillus natto extract.
[0016] Furthermore, the raw materials of the supplement include 3-4wt% of glucose, 0.8-1wt% of vitamin C and 0.3-0.7wt% of dipotassium hydrogen phosphate based on the extract.
[0017] Furthermore, the inoculation refers to 0.8-1.1×10 7 The natto Bacillus subtilis with CFU / g is inoculated into soybean liquid at an inoculation amount of 6-10wt%; the fermentation condition is culturing at 35-40°C for 30-42h; the rotation speed and time of the centrifugation are 8000-12000rmp and 15-25min respectively.
[0018] Beneficial effects of the present invention:
[0019] (1) The present invention utilizes the fermentation liquid of bifidobacterium and lactobacillus strains to inoculate into American ginseng liquid to obtain American ginseng fermentation liquid. The American ginseng liquid is rich in a variety of beneficial ingredients, which provides a suitable nutritional environment for the growth of the strains, promotes the good growth and metabolism of the strains, and thus improves the conversion rate and stability of ginsenosides.
[0020] (2) The American ginseng liquid of the present invention contains American ginseng extract and natto Bacillus extract. The American ginseng extract adopts high-pressure microfluidization treatment, reflux and concentration treatment during the preparation process, which can destroy the cell wall of American ginseng as much as possible, increase the dissolution of effective ingredients, and thus improve the purity and extraction rate of ginsenosides; the natto Bacillus extract obtained by fermentation with soybean extract and supplement as the base provides additional nitrogen source and rich nutrients for the growth of natto Bacillus to a certain extent, promotes the reproduction and metabolic activity of natto Bacillus, and during the fermentation process, natto Bacillus can produce a variety of beneficial metabolites, such as vitamin K2, which promotes the growth and metabolism of probiotics, further improves the fermentation rate, and thus promotes the increase of ginsenoside extraction; in addition, vitamin C is added multiple times during the preparation process of the present invention. Vitamin C has an antioxidant effect and can protect active ingredients such as ginsenosides from being oxidized and destroyed, thereby improving their stability. DETAILED DESCRIPTION
[0021] In order to further illustrate the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the specific implementation methods, structures, features and effects of the present invention are described in detail below in combination with the embodiments.
[0022] Example 1
[0023] A probiotic fermentation process for increasing the content of ginsenosides. The probiotic fermentation process of this embodiment is as follows:
[0024] The fermentation liquid of the strain was inoculated into the American ginseng liquid at an inoculation rate of 10 wt %, the pH was adjusted to 6.2, the liquid was cultured at 30° C. for 48 h, and the American ginseng fermentation liquid was obtained after sterilization at 121° C. for 15 min.
[0025] The concentration of the strain in the strain fermentation broth of this example is 1×10 7 CFU / g, and the strain is composed of bifidobacterium and lactobacillus in a ratio of 1.7:0.8; the fermentation condition of the strain fermentation liquid of this embodiment is anaerobic conditions, cultured at 34°C for 40h, and the fermentation is completed to obtain the strain fermentation liquid.
[0026] The pH of the fermentation medium used for the strain fermentation broth of this embodiment is 6.2, and the raw materials of the fermentation medium include 4.7g / L natto Bacillus extract, 100g / L glucose, 5g / L yeast extract, 1g / L potassium dihydrogen phosphate, 0.3g / L magnesium sulfate, 0.8g / L ferrous sulfate and the balance deionized water.
[0027] The raw materials of the American ginseng liquid of this embodiment include, by weight percentage, 81 wt % of American ginseng extract, 5 wt % of natto Bacillus extract, 1 wt % of vitamin C and the balance of deionized water.
[0028] The preparation method of the American ginseng extract of this embodiment is:
[0029] Wash and dry the American ginseng, grind it through a 200-mesh sieve, add 10 times the weight of deionized water, treat it with high-pressure microfluidization twice, reflux it at 78°C for 1.2h to obtain an extract, and concentrate it by rotary evaporation to obtain the American ginseng extract.
[0030] The pressure and flow rate of the high-pressure microfluidization treatment in this embodiment are 80 MPa and 60 mL / min, respectively.
[0031] The rotary evaporation concentration in this embodiment refers to rotary evaporation at 40° C. until the volume of the original extract is reduced to 1 / 4.
[0032] The preparation method of the Bacillus natto extract of the present embodiment is:
[0033] Soybeans were mixed with deionized water at a solid-liquid ratio of 1:2 g / mL, soaked for 3 hours, drained and steamed in a 100°C steamer for 20 minutes, taken out and cooled, mixed with deionized water at a solid-liquid ratio of 1:4 g / mL, heated to 37°C, stirred at 50 rpm for 1 hour to obtain an extract, added with a supplement, adjusted to pH 6.5, continued stirring to obtain soybean liquid, inoculated, fermented, and centrifuged to obtain the supernatant to obtain a Bacillus natto extract.
[0034] The raw materials of the supplement of this embodiment include 3 wt % of glucose, 0.8 wt % of vitamin C and 0.3 wt % of dipotassium hydrogen phosphate in the extract.
[0035] In this example, the inoculation refers to 0.8×10 7 CFU / g of Bacillus natto was inoculated in soybean liquid at an inoculation rate of 6wt%; the fermentation condition of this example was cultured at 35°C for 30h; the speed and time of centrifugation in this example were 8000rmp and 15min respectively.
[0036] Example 2
[0037] A probiotic fermentation process for increasing the content of ginsenosides. The probiotic fermentation process of this embodiment is as follows:
[0038] The fermentation liquid of the strain was inoculated into the American ginseng liquid at an inoculation rate of 12 wt %, the pH was adjusted to 6.8, the liquid was cultured at 40° C. for 60 h, and the American ginseng fermentation liquid was obtained after sterilization at 121° C. for 15 min.
[0039] The concentration of the strain in the strain fermentation broth of this example is 1.2×10 7 CFU / g, and the strain is composed of bifidobacterium and lactobacillus in a ratio of 2.1:1.2; the fermentation condition of the strain fermentation liquid of this embodiment is anaerobic conditions, cultured at 38°C for 50h, and the fermentation is completed to obtain the strain fermentation liquid.
[0040] The pH of the fermentation medium used for the strain fermentation broth of this embodiment is 6.8, and the raw materials of the fermentation medium include 6.5g / L natto Bacillus extract, 120g / L glucose, 6g / L yeast extract, 2g / L potassium dihydrogen phosphate, 0.7g / L magnesium sulfate, 1g / L ferrous sulfate and the balance deionized water.
[0041] The raw materials of the American ginseng liquid of this embodiment include, by weight percentage, 88 wt % of American ginseng extract, 9 wt % of natto Bacillus extract, 2 wt % of vitamin C and the balance of deionized water.
[0042] The preparation method of the American ginseng extract of this embodiment is:
[0043] Wash and dry the American ginseng, grind it through a 300-mesh sieve, add 16 times the weight of deionized water, treat it with high-pressure microfluidization twice, reflux it at 82°C for 2h to obtain an extract, and concentrate it by rotary evaporation to obtain the American ginseng extract.
[0044] The pressure and flow rate of the high-pressure microfluidization treatment in this embodiment are 120 MPa and 100 mL / min, respectively.
[0045] The rotary evaporation concentration in this embodiment refers to rotary evaporation at 40° C. until the volume of the original extract is reduced to 1 / 4.
[0046] The preparation method of the Bacillus natto extract of the present embodiment is:
[0047] Soybeans were mixed with deionized water at a solid-liquid ratio of 1:6 g / mL, soaked for 5 hours, drained and steamed in a 100°C steamer for 30 minutes, taken out and cooled, mixed with deionized water at a solid-liquid ratio of 1:10 g / mL, heated to 41°C, stirred at 100 rpm for 2 hours to obtain an extract, added with a supplement, adjusted to pH 7.5, continued stirring to obtain soybean liquid, inoculated, fermented, and centrifuged to obtain the supernatant to obtain a Bacillus natto extract.
[0048] The raw materials of the supplement of this embodiment include 4 wt % of glucose, 1 wt % of vitamin C and 0.7 wt % of dipotassium hydrogen phosphate in the extract.
[0049] In this example, the inoculation refers to the addition of 1.1×10 7 CFU / g of Bacillus natto was inoculated into soybean liquid at an inoculation rate of 10wt%; the fermentation condition of this embodiment was cultured at 40°C for 30-42h; the centrifugation speed and time of this embodiment were 12000rmp and 25min respectively.
[0050] Example 3
[0051] A probiotic fermentation process for increasing the content of ginsenosides. The probiotic fermentation process of this embodiment is as follows:
[0052] The fermentation liquid of the strain was inoculated into the American ginseng liquid at an inoculation rate of 11 wt %, the pH was adjusted to 6.5, the liquid was cultured at 35° C. for 54 h, and the American ginseng fermentation liquid was obtained after sterilization at 121° C. for 15 min.
[0053] The concentration of the strain in the strain fermentation broth of this example is 1.1×10 7 CFU / g, and the strain is composed of bifidobacterium and lactobacillus in a ratio of 1.9:1; the fermentation condition of the strain fermentation liquid of this embodiment is anaerobic conditions, cultured at 36°C for 45h, and the fermentation is completed to obtain the strain fermentation liquid.
[0054] The pH of the fermentation medium used for the strain fermentation broth of this embodiment is 6.5, and the raw materials of the fermentation medium include 5.6g / L natto Bacillus extract, 110g / L glucose, 5.5g / L yeast extract, 1.5g / L potassium dihydrogen phosphate, 0.5g / L magnesium sulfate, 0.9g / L ferrous sulfate and the balance deionized water.
[0055] The raw materials of the American ginseng liquid of this embodiment include, by weight percentage, 84.5 wt % of American ginseng extract, 7 wt % of natto Bacillus extract, 1.5 wt % of vitamin C and the balance of deionized water.
[0056] The preparation method of the American ginseng extract of this embodiment is:
[0057] Wash and dry the American ginseng, grind it through a 250-mesh sieve, add 13 times the weight of deionized water, treat it with high-pressure microfluidization twice, reflux it at 80°C for 1.6 hours to obtain an extract, and concentrate it by rotary evaporation to obtain the American ginseng extract.
[0058] The pressure and flow rate of the high-pressure microfluidization treatment in this embodiment are 100 MPa and 80 mL / min, respectively.
[0059] The rotary evaporation concentration in this embodiment refers to rotary evaporation at 40° C. until the volume of the original extract is reduced to 1 / 4.
[0060] The preparation method of the Bacillus natto extract of the present embodiment is:
[0061] Soybeans were mixed with deionized water at a solid-liquid ratio of 1:4 g / mL, soaked for 4 hours, drained and steamed in a 100°C steamer for 25 minutes, taken out and cooled, mixed with deionized water at a solid-liquid ratio of 1:7 g / mL, heated to 39°C, stirred at 75 rpm for 1.5 hours to obtain an extract, added a supplement, adjusted the pH to 7, continued stirring to obtain soybean liquid, inoculated, fermented, and centrifuged to obtain the supernatant to obtain a Bacillus natto extract.
[0062] The raw materials of the supplement of this embodiment include 3.5 wt % of glucose, 0.9 wt % of vitamin C and 0.5 wt % of dipotassium hydrogen phosphate in the extract.
[0063] In this example, the inoculation refers to 0.95×10 7 CFU / g of Bacillus natto was inoculated in soybean liquid at an inoculation rate of 8wt%; the fermentation condition of this example was cultured at 37.5°C for 36h; the speed and time of centrifugation in this example were 10000rmp and 20min respectively.
[0064] Comparative Example 1
[0065] On the basis of Example 3, the American ginseng liquid was removed and replaced with an equal weight of American ginseng extract, and other conditions were consistent with Example 3.
[0066] Comparative Example 2
[0067] On the basis of Example 3, the Bacillus natto extract in the American ginseng solution was removed and replaced with an equal weight of deionized water. Other conditions were the same as those in Example 3.
[0068] Comparative Example 3
[0069] On the basis of Example 3, the Bacillus natto extract in the fermentation medium of the strain fermentation liquid was removed and replaced with an equal weight of deionized water. Other conditions were the same as those in Example 3.
[0070] Taking the ginsenoside content of the control group as a benchmark, the American ginseng fermentation broth obtained in Examples 1-3 and Comparative Examples 1-3 was used as a sample, and the test was performed using an Agilent 1260 chromatograph, a ZORBAX SB-C18 (4.6 mm×250 mm, 5 μm) chromatographic column, the column temperature was set to 35°C, the column flow rate was 0.8 mL / min, the injection volume was 5 μL, 0.2% acetic acid water was used as mobile phase A, and acetonitrile was used as mobile phase B to detect the increase in the amount of ginsenosides.
[0071] The extraction method of ginsenosides in the control group was:
[0072] Wash and dry the American ginseng, grind it through a 250-mesh sieve, add 13 times the weight of deionized water, reflux at 80°C for 1.6 hours to obtain an extract, and concentrate it by rotary evaporation to 1 / 4 of the original extract volume to obtain the American ginseng extract.
[0073] Table 1
[0074] Increased amount of ginsenosides% Control group 0 Example 1 1.92 Example 2 1.93 Example 3 1.96 Comparative Example 1 0.35 Comparative Example 2 1.66 Comparative Example 3 1.43
[0075] As can be seen from Table 1, the ginsenoside content in Examples 1-3 of the present invention is improved compared with conventional alcohol extraction and reflux treatment. Although Comparative Example 1 directly uses American ginseng extract instead of American ginseng liquid, it also uses high-pressure microfluidization treatment during the extraction process, which can more effectively extract ginsenosides in American ginseng; Comparative Examples 2 and 3 remove the natto Bacillus extract in the corresponding steps. The natto Bacillus extract provides an additional nitrogen source and rich nutrients for the growth of Bacillus natto to a certain extent, thereby promoting the growth and metabolism of bifidobacteria and lactobacilli, and has a certain promoting effect on the extraction and conversion of ginsenosides, so the extraction amount of ginsenosides is reduced.
[0076] The above description is only a preferred embodiment of the present invention and does not limit the present invention in any form. Although the present invention has been disclosed as a preferred embodiment as above, it is not used to limit the present invention. Any technical personnel in this field can make some changes or modify the technical contents disclosed above into equivalent embodiments without departing from the scope of the technical solution of the present invention. However, any indirect modification, equivalent change and modification made to the above embodiments according to the technical essence of the present invention without departing from the content of the technical solution of the present invention still fall within the scope of the technical solution of the present invention.
Claims
1. A probiotic fermentation process for increasing the content of ginsenosides, characterized in that: The probiotic fermentation process is: The fermentation liquid of the strain is inoculated into the American ginseng liquid at an inoculation rate of 10-12wt%, the pH is adjusted to 6.2-6.8, the liquid is cultured at 30-40°C for 48-60h, and the American ginseng fermentation liquid is obtained after sterilization at 121°C for 15min.
2. The probiotic fermentation process for increasing the content of ginsenosides according to claim 1, characterized in that: The concentration of the strain in the fermentation broth is 1-1.2×10 7 CFU / g, and the strain is composed of bifidobacterium and lactobacillus in a ratio of 1.7-2.1:0.8-1.2; the fermentation condition of the strain fermentation liquid is anaerobic conditions, cultured at 34-38°C for 40-50h, and the fermentation is completed to obtain the strain fermentation liquid.
3. The probiotic fermentation process for increasing the content of ginsenosides according to claim 2, characterized in that: The pH of the fermentation medium used for the strain fermentation liquid is 6.2-6.8, and the raw materials of the fermentation medium include 4.7-6.5 g / L natto extract, 100-120 g / L glucose, 5-6 g / L yeast extract, 1-2 g / L potassium dihydrogen phosphate, 0.3-0.7 g / L magnesium sulfate, 0.8-1 g / L ferrous sulfate and the balance of deionized water.
4. The probiotic fermentation process for increasing the content of ginsenosides according to claim 1, characterized in that: The raw materials of the American ginseng liquid include, by mass percentage, 81-88wt% of American ginseng extract, 5-9wt% of natto bacteria extract, 1-2wt% of vitamin C and the balance of deionized water.
5. The probiotic fermentation process for increasing the content of ginsenosides according to claim 4, characterized in that: The preparation method of the American ginseng extract is: Wash and dry the American ginseng, crush it through a 200-300 mesh sieve, add 10-16 times the weight of deionized water, treat it with high-pressure microfluidization twice, reflux it at 78-82°C for 1.2-2h to obtain an extract, and concentrate it by rotary evaporation to obtain the American ginseng extract.
6. A probiotic fermentation process for increasing the content of ginsenosides according to claim 5, characterized in that: The pressure and flow rate of the high-pressure microfluidization treatment are 80-120 MPa and 60-100 mL / min, respectively.
7. The probiotic fermentation process for increasing the content of ginsenosides according to claim 5, characterized in that: The rotary evaporation concentration refers to rotary evaporation at 40° C. to reduce the volume of the original extract to 1 / 4.
8. A probiotic fermentation process for increasing the content of ginsenosides according to claim 3 or 4, characterized in that: The preparation method of the Bacillus natto extract is: The soybeans were mixed with deionized water at a solid-liquid ratio of 1:2-6 g / mL, soaked for 3-5 hours, drained and steamed in a 100°C steamer for 20-30 minutes, taken out and cooled, mixed with deionized water at a solid-liquid ratio of 1:4-10 g / mL, heated to 37-41°C, stirred at 50-100 rmp for 1-2 hours to obtain an extract, added with a supplement, adjusted to pH 6.5-7.5, continued stirring to obtain soybean liquid, inoculated, fermented, centrifuged and obtained a supernatant to obtain a Bacillus natto extract.
9. A probiotic fermentation process for increasing the content of ginsenosides according to claim 8, characterized in that: The raw materials of the supplement include 3-4wt% of glucose, 0.8-1wt% of vitamin C and 0.3-0.7wt% of dipotassium hydrogen phosphate in the extract.
10. The probiotic fermentation process for increasing the content of ginsenosides according to claim 8, characterized in that: The inoculation refers to 0.8-1.1×10 7 The natto Bacillus subtilis with CFU / g is inoculated into soybean liquid at an inoculation amount of 6-10wt%; the fermentation condition is culturing at 35-40°C for 30-42h; the rotation speed and time of the centrifugation are 8000-12000rmp and 15-25min respectively.