A method for increasing arginine glycosides in red ginseng extract and American ginseng extract

Abstract

The present invention provides a method for increasing arginine glycosides in red ginseng extract and American ginseng extract, and the specific steps include ginseng crushing and screening, composite strain fermentation, hot reflux extraction, and reduced pressure concentration, wherein the composite strain fermentation adopts licheniformis and Bacillus subtilis for cooperative fermentation, metabolizes and decomposes protein, starch, etc., produces arginine and reducing sugar, and provides a good reaction substrate basis for the generation of arginine glycosides, and the high reaction substrate basis can induce the Maillard reaction to proceed in the reaction direction of carbonyl aldehyde condensation, and reduce the influence of Maillard reaction byproducts on arginine glycosides. In addition, the application of catechins can inhibit the Maillard reaction of other amino-containing substances and carbonyl substances in the subsequent heating extraction step, reduce the influence of the final product of the Maillard reaction on the quality, ensure the excellent quality of red ginseng extract and American ginseng extract, and provide solid technical support for the sustainable development of the health care product market.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine processing, and particularly relates to a method for increasing arginine glycoside in red ginseng extract and American ginseng extract. Background Art

[0002] American ginseng extract and red ginseng extract are both precious health products derived from ginseng plants, each with unique effects and characteristics. American ginseng extract is refined from the dried roots of American ginseng, a plant of the Araliaceae family, through modern extraction technology. It is rich in saponins, arginine glycosides, fatty acids, sterols and other active ingredients, giving American ginseng extract a variety of health benefits such as replenishing qi and nourishing yin, clearing away heat and promoting fluid production, anti-fatigue, anti-aging, and enhancing immunity; while red ginseng extract is made by processing ginseng through a complex process into red ginseng, and then further processed into an extract. It not only retains the original nutrients of red ginseng such as ginsenosides, arginine glycosides, polyphenols, flavonoids, etc., but also gives red ginseng extract good medicinal effects, such as replenishing qi and blood, nourishing yin and promoting fluid production, strengthening the heart and spleen, calming the nerves, etc., and makes it easier for the human body to absorb in the form of an extract, effectively regulating the immune and endocrine systems, and promoting physical recovery.

[0003] Arginine glycoside is an arginine derivative, including arginine disaccharide and arginine monoglycoside, which is the intermediate product of the Maillard reaction between arginine and maltose during the steaming and drying process of fresh ginseng processed into red ginseng. Studies have shown that amino acid derivatives are very rich in red ginseng, up to 4% or more, and have significant effects in anti-oxidation, dilation of blood vessels, promotion of microcirculation, enhancement of immunity, anti-diabetes, inhibition of tyrosinase activity, inhibition of melanin production, and inhibition of arginine protein nitrogen methyltransferase activity.

[0004] In the prior art, arginine glycosides can be artificially synthesized by providing arginine and reducing sugar substrates. However, the biological activities of artificially synthesized arginine glycosides are somewhat different from those of natural arginine glycosides in red ginseng extract and American ginseng extract. Their effects may not be as stable and reliable as those of natural arginine glycosides. They usually coexist with multiple other active ingredients to have a highly efficient synergistic effect, while artificially synthesized products are often only a single ingredient, and it is difficult to completely simulate this natural advantage.

[0005] However, during the processing of red ginseng extract and American ginseng extract, red ginseng and American ginseng contain other ingredients besides Maillard reaction substrates such as protein and starch, which will induce the main direction of the Maillard reaction and increase the probability of Maillard side reactions. At the same time, arginine glycosides are generated in the early stage of the Maillard reaction, and there is an excessive Maillard reaction later to form a late glycosylation final product, which will significantly reduce the content of arginine glycosides and produce harmful substances, affecting the quality of red ginseng extract and American ginseng extract. Therefore, it is urgent to provide a method for increasing arginine glycosides in red ginseng extract and American ginseng extract, so as to provide solid technical support and quality assurance for the sustainable development of the health care product market such as red ginseng extract and American ginseng extract. Summary of the invention

[0006] Technical problems to be solved: In view of the above technical problems, the purpose of the present invention is to provide a method for increasing the arginine glycosides in red ginseng extract and American ginseng extract, the specific steps include ginseng crushing and screening, composite strain fermentation, hot reflux extraction, and reduced pressure concentration, wherein the composite strain fermentation uses Bacillus licheniformis and Bacillus subtilis for synergistic fermentation, metabolizes and decomposes proteins, starch, etc., produces arginine and reducing sugars, and provides a good reaction substrate basis for the generation of arginine glycosides, and the high reaction substrate basis can induce the Maillard reaction to proceed in the direction of carbonyl aldehyde condensation, reducing the influence of Maillard reaction byproducts on arginine glycosides. In addition, the application of catechins can inhibit the Maillard reaction of other amino-containing substances and carbonyl substances in the subsequent heating extraction step, reduce the influence of the final product of the Maillard reaction on the quality, ensure the excellent quality of red ginseng extract and American ginseng extract, and provide solid technical support for the sustainable development of the health care product market.

[0007] Technical solution: A method for increasing arginine glycoside in red ginseng extract and American ginseng extract, comprising the following steps: S1. selecting ginseng of good quality, crushing and passing through a 50-80 mesh sieve to obtain ginseng powder;

[0008] S2. Ginseng powder was added with water at a solid-liquid ratio of 1: (2-5) and stirred to obtain a ginseng fermentation matrix;

[0009] S3. Inoculate 2-8% of the composite bacterial solution in the ginseng fermentation matrix and ferment at 30-40 ° C for 12-48h to obtain ginseng fermentation liquid;

[0010] S4. The ginseng fermentation liquid was heated at 50-100 ° C for 1-3 h, and then ethanol was added at a solid-liquid ratio of 1: (2-5) and refluxed for 1-3 h to obtain an alcohol-extracted ginseng fermentation liquid;

[0011] S5. The alcohol-extracted ginseng fermentation liquid is mixed with water at a solid-liquid ratio of 1:(5-10), and then 0.01-0.04% catechin is added, and the mixture is subjected to hot reflux extraction for 1-3 hours, and concentrated under reduced pressure to obtain a ginseng extract.

[0012] Furthermore, the ginseng in step S1 includes red ginseng and American ginseng.

[0013] Furthermore, in step S3, the composite bacterial species are Bacillus licheniformis and Bacillus subtilis, and the ratio of Bacillus licheniformis to Bacillus subtilis is (3-1):(1-3).

[0014] Furthermore, the heating temperature of the ethanol reflux extraction in step S4 is 60-80° C., and the number of extractions is 1-3 times.

[0015] Furthermore, the heating temperature of the hot reflux extraction in step S5 is 60-100° C., and the number of extractions is 1-3 times.

[0016] Furthermore, the activation method of Bacillus licheniformis comprises the following steps:

[0017] Step 1. Thaw the frozen Bacillus licheniformis and inoculate it into the sterilized LB solid medium, and culture it overnight at 28-32°C until colonies grow;

[0018] Step 2. Pick a single colony of Bacillus licheniformis with an inoculation loop and inoculate it into a sterilized LB liquid culture medium, and culture it at 28-32° C. and 150-200 rpm for 12-15 hours to obtain a Bacillus licheniformis seed solution;

[0019] Step 3: Inoculate 1-5% of Bacillus licheniformis seed solution in the fermentation medium. The initial strain concentration of the fermentation medium is (2-4)×10 5 CFU / mL, and cultured at 28-32℃, 150-200rpm for 24h to obtain activated Bacillus licheniformis bacterial solution, the concentration of which was (5-8)×10 8 CFU / mL.

[0020] Furthermore, the activation method of Bacillus subtilis comprises the following steps:

[0021] Step 4. Thaw the frozen Bacillus subtilis and inoculate it into the sterilized LB solid medium, and culture it overnight at 32-37°C until colonies grow;

[0022] Step 5. Pick a single colony of Bacillus subtilis with an inoculation loop and inoculate it into a sterilized LB liquid culture medium, and culture it at 32-37° C. and 120-180 rpm for 12-15 hours to obtain a Bacillus subtilis seed solution;

[0023] Step 6. Inoculate 1-5% of Bacillus subtilis seed solution in the fermentation medium. The initial strain concentration of the fermentation medium is (3-4)×10 5CFU / mL, and cultured at 32-37°C and 150-200rpm for 24h to obtain activated Bacillus subtilis culture solution with a concentration of (7-9)×10 8 CFU / mL.

[0024] Beneficial effects:

[0025] 1. The present invention utilizes Bacillus licheniformis and Bacillus subtilis for cooperative fermentation. Bacillus subtilis can produce amylase, cellulase and protease, etc., decompose starch, cellulose and protein in red ginseng and American ginseng, produce reducing sugar and polypeptide, increase the reducing sugar content in the reaction substrate, and provide more easily available nutrients for Bacillus licheniformis. At the same time, Bacillus licheniformis utilizes the polypeptide in the decomposed substrate to produce arginine by fermentation, increase the arginine content in the reaction system, and provide a good substrate basis for the synthesis of arginine glycosides. In addition, in the cooperative fermentation system of Bacillus licheniformis and Bacillus subtilis, the presence of Bacillus licheniformis helps to regulate the oxygen concentration in the fermentation environment, create a more suitable aerobic environment for Bacillus subtilis, and the two jointly regulate the environmental conditions of the fermentation system, so that the fermentation environment is more stable and the growth and metabolism of the two are promoted.

[0026] 2. The high reaction substrate content of arginine glycoside in the present invention can induce the Maillard reaction to proceed in the direction of carbonyl aldehyde condensation, reduce the occurrence of Maillard reaction side reactions, and thus reduce the influence of Maillard reaction side reaction products on arginine glycoside. At the same time, the addition of catechins can prevent other amino-containing substances and carbonyl substances from undergoing Maillard reaction during the preparation of red ginseng extract and American ginseng extract, generate Maillard reaction end products, and ensure the high quality and arginine glycoside content of red ginseng extract and American ginseng extract.

[0027] 3. The present invention increases the content of arginine glycoside in red ginseng extract and American ginseng extract based on the composite fermentation of microbial strains, which not only can give red ginseng extract and American ginseng extract good physiological activity and functional utility, but also promotes the improvement of production technology and the improvement of product quality, providing new ideas and methods for the modernization and industrialization of ginseng. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with embodiments, which are explanations of the present invention and are not limited to the following embodiments:

[0029] The strains used in this patent: Bacillus licheniformis CICC22589 and Bacillus subtilis CICC24534, were purchased from the China Industrial Microbiological Culture Collection Center.

[0030] Example 1

[0031] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0032] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0033] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng fermentation matrix;

[0034] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0035] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0036] S5. Add 8.4 L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.36 g of catechins and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0037] Example 2

[0038] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0039] S1. Select 300g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0040] S2. Weigh 200 g of red ginseng powder, add 600 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0041] S3. Inoculate 40 mL of composite bacterial solution (20 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0042] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 3.2L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0043] S5. Add 5.6L of water to the alcohol-extracted red ginseng fermentation liquid, and then add 0.24g of catechins. Extract under hot reflux at 80°C for 2h, repeat twice, and concentrate under reduced pressure to obtain red ginseng extract.

[0044] Example 3

[0045] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0046] S1. Select 500g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0047] S2. Weigh 400 g of red ginseng powder, add 1600 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0048] S3. Inoculate 80 mL of composite bacterial solution (26.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8 CFU / mL, 53.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0049] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 6.4L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0050] S5. Add 11.2L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.48g of catechins and extract it under hot reflux at 80°C for 2h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0051] Example 4

[0052] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0053] S1. Select 450g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0054] S2. Weigh 350 g of red ginseng powder, add 1400 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0055] S3. Inoculate 28 mL of composite bacterial solution (18.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 9.3mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0056] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 5.6L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0057] S5. Add 9.8 L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.42 g of catechins and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0058] Example 5

[0059] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0060] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0061] S2. Weigh 200 g of red ginseng powder, add 600 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0062] S3. Inoculate 64 mL of composite bacterial solution (42.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8 CFU / mL, 21.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0063] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 3.2L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0064] S5. Add 5.6L of water to the alcohol-extracted red ginseng fermentation liquid, and then add 0.24g of catechins. Extract under hot reflux at 80°C for 2h, repeat twice, and concentrate under reduced pressure to obtain red ginseng extract.

[0065] Example 6

[0066] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0067] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0068] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng fermentation matrix;

[0069] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×108 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0070] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0071] S5. Add 8.4 L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.12 g of catechins and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0072] Example 7

[0073] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0074] S1. Select 500g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0075] S2. Weigh 400 g of red ginseng powder, add 1600 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0076] S3. Inoculate 80 mL of composite bacterial solution (53.3 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 26.7mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0077] S4. The red ginseng fermentation liquid was heated at 50°C for 2h, and then 6.4L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0078] S5. Add 11.2L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.48g of catechins and extract it under hot reflux at 80°C for 2h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0079] Example 8

[0080] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0081] S1. Select 450g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0082] S2. Weigh 350 g of red ginseng powder, add 1400 mL of water and stir to mix evenly to obtain a red ginseng fermentation matrix;

[0083] S3. Inoculate 70 mL of composite bacterial solution (46.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8 CFU / mL, 23.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0084] S4. The red ginseng fermentation liquid was heated at 100°C for 2h, and then 5.6L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0085] S5. Add 9.8L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.42g of catechins and extract it under hot reflux at 80°C for 2h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0086] Comparative Example 1

[0087] The difference between this comparative example and Example 1 is that Bacillus subtilis is replaced by Bacillus licheniformis.

[0088] A method for increasing arginine glycoside in red ginseng extract by using Bacillus licheniformis, comprising the following steps:

[0089] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0090] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng fermentation matrix;

[0091] S3. Inoculate 60 mL of Bacillus licheniformis solution (concentration of the solution was 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0092] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0093] S5. Add 8.4 L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.36 g of catechins and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0094] Comparative Example 2

[0095] The difference between this comparative example and Example 1 is that Bacillus licheniformis is replaced by Bacillus subtilis.

[0096] A method for increasing arginine glycoside in red ginseng extract by using Bacillus subtilis comprises the following steps:

[0097] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0098] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng fermentation matrix;

[0099] S3. Inoculate 60 mL of Bacillus subtilis solution (concentration of the solution was 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0100] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0101] S5. Add 8.4 L of water to the alcohol-extracted red ginseng fermentation liquid and mix it. Then add 0.36 g of catechins and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0102] Comparative Example 3

[0103] The difference between this comparative example and Example 1 is that catechin is not added.

[0104] A method for increasing arginine glycoside in red ginseng extract, comprising the following steps:

[0105] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0106] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng fermentation matrix;

[0107] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain red ginseng fermentation liquid;

[0108] S4. The red ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted red ginseng fermentation liquid;

[0109] S5. Add 8.4 L of water to the alcohol-extracted red ginseng fermentation liquid, mix it, and extract it under hot reflux at 80°C for 2 h. Repeat the cycle twice, and concentrate under reduced pressure to obtain red ginseng extract.

[0110] Comparative Example 4

[0111] The difference between this comparative example and Example 1 is that the composite bacterial liquid fermentation is replaced by enzymatic hydrolysis, wherein the enzymatic hydrolysis adopts cellulase (enzyme activity 15 U / mg) and medium-temperature α-amylase (enzyme activity 50 U / mg).

[0112] A method for increasing arginine glycosides in red ginseng extract based on enzymatic hydrolysis, comprising the following steps:

[0113] S1. Select 400g of good quality red ginseng, crush and pass through an 80-mesh sieve to obtain red ginseng powder;

[0114] S2. Weigh 300 g of red ginseng powder, add 900 mL of water and stir to mix well to obtain a red ginseng solution;

[0115] S3. Add 6g of complex enzyme (3g of cellulase, 3g of medium-temperature α-amylase) to the red ginseng solution, enzymatically hydrolyze for 3h at pH 6.0 and 55°C, and inactivate the enzyme at high temperature to obtain a red ginseng enzymatic solution;

[0116] S4. The red ginseng hydrolysate was heated at 80°C for 2h, and then 4.8L of ethanol was added and heated under reflux extraction at 75°C for 2h. This cycle was repeated twice to obtain an alcohol-extracted red ginseng hydrolysate.

[0117] S5. Add 8.4 L of water to the alcohol-extracted red ginseng hydrolysate and mix. Then add 0.36 g of catechins and perform hot reflux extraction at 80 °C for 2 h. Repeat the cycle twice and concentrate under reduced pressure to obtain red ginseng extract.

[0118] Performance Testing

[0119] (1) Arginine glycoside content

[0120] The contents of arginine monoglucoside and arginine disoside in the examples and comparative examples were determined by high performance liquid chromatography-ultraviolet visible absorption detector method (HPLC-UV).

[0121] The test conditions are shown in Table 1.

[0122] Table 1 HPLC-UV analysis conditions

[0123] project condition Chromatographic columns Venusil-AA amino acid analysis column Column specifications Particle size 5μm, size 4.6mm×250mm Column temperature 40℃ Detection wavelength 254nm Mobile phase A Sodium acetate buffer solution (pH 6.5)-acetonitrile solution Mobile phase B Acetonitrile-water (4:1, v / v) solution Flow rate 1.0mL / min Linear gradient elution program 0min, 0%B; 4min, 3%B; 16min, 10%B

[0124] Preparation and derivatization of AF standard: Accurately weigh 0.015 g of AF standard and dissolve it in water to make a volume of 10 mL. Take 1 mL and place it in a 2 mL centrifuge tube. Add 100 μL of triethylamine acetonitrile solution and 50 μL of phenyl isothiocyanate acetonitrile solution, mix well, and let it stand at room temperature for 1 hour. Then add 400 μL of n-hexane, shake and let it stand for 10 minutes. Take the lower layer of solution, filter it with a 0.2 μm filter membrane, and take 20 μL for chromatographic analysis.

[0125] Preparation and derivatization of AFG standard: Accurately weigh 0.015 g of AFG standard and dissolve it in water to make a volume of 10 mL. Take 1 mL and place it in a 2 mL centrifuge tube. The following derivatization steps are the same as above. Let it stand at room temperature for 1 hour, then add 400 μL of n-hexane, shake it and let it stand for 10 minutes, take the lower layer of solution, filter it with a 0.2 μm filter membrane, and take 20 μL for chromatographic analysis.

[0126] Sample preparation and derivatization: Accurately weigh 1.0g of sample, add 20mL of methanol to soak overnight, filter, add 20mL of water to the residue, extract 3 times (250W, 100kHZ), 30min each time, filter, combine the filtrate, low pressure rotary concentration to dryness, dissolve in distilled water and transfer to a 10mL volumetric flask, and make up to volume. Use a pipette to take 1mL and add it to a 2mL centrifuge tube, add 100μL of triethylamine acetonitrile solution and 100μL of phenyl isothiocyanate acetonitrile solution, mix well, place at room temperature for 1h, then add 400μL of n-hexane, shake evenly and place for 10min, take the lower layer of solution, filter with a 0.2μm filter membrane, and take 20μL for injection analysis.

[0127] Table 2 Arginine monoglycoside and arginine disaccharide content of Examples 1-8 and Comparative Examples 1-4

[0128] Arginine monoglycoside (%) Arginine disodium glycoside (%) Arginine glycoside (%) Example 1 3.57 9.81 13.38 Example 2 3.30 9.01 12.31 Example 3 3.20 8.99 12.19 Example 4 3.18 8.89 12.07 Example 5 3.33 9.18 12.51 Example 6 3.24 8.99 12.23 Example 7 3.36 9.13 12.48 Example 8 3.26 9.08 12.34 Comparative Example 1 2.87 8.24 11.12 Comparative Example 2 2.95 8.17 11.12 Comparative Example 3 3.00 8.57 11.56 Comparative Example 4 2.58 7.90 10.48

[0129] As can be seen from Table 2, the total amount of arginine monoglycoside, arginine disaccharide and arginine glycoside in Examples 1-8 is higher than that in Comparative Examples 1-4, indicating that the composite bacterial fermentation can act on the protein, starch and cellulose in red ginseng, metabolize and decompose to generate arginine glycoside synthesis substrates, and the high content of arginine and reducing sugar specifically guides the direction of the Maillard reaction, and specifically increases the content of arginine glycoside in the red ginseng extract, and catechins can inhibit the occurrence of the Maillard reaction in the subsequent heating reflux extraction process, prevent the excessive degree of the Maillard main reaction, thereby generating other Maillard by-products, affecting the quality of arginine glycosides, thereby ensuring the high quality of the red ginseng extract; in addition, Comparative Example 4 uses an enzymatic hydrolysis method to increase the content of arginine glycoside in the red ginseng extract, and the effect of the embodiment is better than that of Comparative Example 4, which further illustrates that compared with the addition of a single-function enzyme, the multiple enzyme systems produced by microbial fermentation can decompose various components in red ginseng to the greatest extent, not only providing nutrients for microbial growth, but also increasing the content of arginine glycoside synthesis substrates, which is beneficial to the synthesis of arginine glycosides in red ginseng extract.

[0130] Example 9

[0131] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0132] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0133] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0134] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0135] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0136] S5. Add 8.4 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.36 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0137] Example 10

[0138] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0139] S1. Select 300g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0140] S2. Weigh 200 g of American ginseng powder, add 600 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0141] S3. Inoculate 40 mL of composite bacterial solution (20 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0142] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 3.2L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0143] S5. Add 5.6L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.24g of catechins. Extract under hot reflux at 80°C for 2h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0144] Embodiment 11

[0145] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0146] S1. Select 500g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0147] S2. Weigh 400 g of American ginseng powder, add 1600 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0148] S3. Inoculate 80 mL of composite bacterial solution (26.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8 CFU / mL, 53.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0149] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 6.4L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0150] S5. Add 11.2 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.48 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0151] Example 12

[0152] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0153] S1. Select 450g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0154] S2. Weigh 350 g of American ginseng powder, add 1400 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0155] S3. Inoculate 28 mL of composite bacterial solution (18.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 9.3mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0156] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 5.6L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0157] S5. Add 9.8 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.42 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0158] Example 13

[0159] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0160] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0161] S2. Weigh 200 g of American ginseng powder, add 600 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0162] S3. Inoculate 64 mL of composite bacterial solution (42.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8 CFU / mL, 21.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0163] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 3.2L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0164] S5. Add 5.6L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.24g of catechins. Extract under hot reflux at 80°C for 2h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0165] Embodiment 14

[0166] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0167] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0168] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0169] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0170] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0171] S5. Add 8.4 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.12 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0172] Embodiment 15

[0173] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0174] S1. Select 500g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0175] S2. Weigh 400 g of American ginseng powder, add 1600 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0176] S3. Inoculate 80 mL of composite bacterial solution (53.3 mL of Bacillus licheniformis solution, bacterial solution concentration 7.5×10 8 CFU / mL, 26.7mL Bacillus subtilis solution, bacterial concentration 7.5×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0177] S4. The American ginseng fermentation liquid was heated at 50°C for 2h, and then 6.4L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0178] S5. Add 11.2 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.48 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0179] Example 16

[0180] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0181] S1. Select 450g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0182] S2. Weigh 350 g of American ginseng powder, add 1400 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0183] S3. Inoculate 70 mL of composite bacterial solution (46.7 mL of Bacillus licheniformis solution, bacterial solution concentration 7×10 8CFU / mL, 23.3mL Bacillus subtilis solution, bacterial concentration 7×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0184] S4. The American ginseng fermentation liquid was heated at 100°C for 2h, and then 5.6L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0185] S5. Add 9.8 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.42 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0186] Comparative Example 5

[0187] The difference between this comparative example and Example 9 is that Bacillus subtilis is replaced by Bacillus licheniformis.

[0188] A method for increasing arginine glycoside in American ginseng extract by using Bacillus licheniformis comprises the following steps:

[0189] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0190] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0191] S3. Inoculate 60 mL of Bacillus licheniformis solution (concentration of bacterial solution 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0192] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0193] S5. Add 8.4 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.36 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0194] Comparative Example 6

[0195] The difference between this comparative example and Example 9 is that Bacillus licheniformis is replaced by Bacillus subtilis.

[0196] A method for increasing arginine glycoside in American ginseng extract by using Bacillus subtilis comprises the following steps:

[0197] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0198] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0199] S3. Inoculate 60 mL of Bacillus subtilis solution (concentration of the solution was 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0200] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0201] S5. Add 8.4 L of water to the alcohol-extracted American ginseng fermentation liquid, and then add 0.36 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0202] Comparative Example 7

[0203] The difference between this comparative example and Example 9 is that catechin is not added.

[0204] A method for increasing arginine glycoside in an American ginseng extract comprises the following steps:

[0205] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0206] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix evenly to obtain an American ginseng fermentation matrix;

[0207] S3. Inoculate 60 mL of composite bacterial solution (40 mL of Bacillus licheniformis solution, bacterial solution concentration 8×10 8 CFU / mL, 20mL Bacillus subtilis solution, bacterial concentration 8×10 8 CFU / mL), and fermented at 35°C for 24h to obtain American ginseng fermentation liquid;

[0208] S4. The American ginseng fermentation liquid was heated at 80°C for 2h, and then 4.8L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2h, and the cycle was repeated twice to obtain an alcohol-extracted American ginseng fermentation liquid;

[0209] S5. Add 8.4 L of water to the alcohol-extracted American ginseng fermentation liquid, mix it, and extract it under hot reflux at 80°C for 2 h, repeat it twice, and concentrate it under reduced pressure to obtain the American ginseng extract.

[0210] Comparative Example 8

[0211] The difference between this comparative example and Example 9 is that the composite bacterial liquid fermentation is replaced by enzymatic hydrolysis treatment, wherein the enzymatic hydrolysis treatment adopts cellulase (enzyme activity 15 U / mg) and medium-temperature α-amylase (enzyme activity 50 U / mg).

[0212] A method for increasing arginine glycosides in American ginseng extract based on enzymatic hydrolysis, comprising the following steps:

[0213] S1. Select 400g of good quality American ginseng, crush and pass through an 80-mesh sieve to obtain American ginseng powder;

[0214] S2. Weigh 300 g of American ginseng powder, add 900 mL of water and stir to mix well to obtain an American ginseng solution;

[0215] S3. Add 6g of complex enzyme (3g of cellulase, 3g of medium-temperature α-amylase) to the American ginseng solution, enzymatically hydrolyze for 3h at pH 6.0 and 55°C, and inactivate the enzyme at high temperature to obtain an American ginseng hydrolyzate;

[0216] S4. The American ginseng hydrolysate was heated at 80°C for 2 hours, and then 4.8 L of ethanol was added, and the mixture was heated under reflux extraction at 75°C for 2 hours. The cycle was repeated twice to obtain an alcohol-extracted American ginseng hydrolysate.

[0217] S5. Add 8.4 L of water to the alcohol-extracted American ginseng hydrolysate, and then add 0.36 g of catechins. Extract under hot reflux at 80°C for 2 h, repeat twice, and concentrate under reduced pressure to obtain American ginseng extract.

[0218] Table 3 Arginine monoglycoside and arginine disaccharide content of Examples 9-16 and Comparative Examples 5-8

[0219] Arginine monoglycoside (%) Arginine disodium glycoside (%) Arginine glycoside (%) Example 9 0.949 0.570 1.518 Example 10 0.929 0.546 1.475 Embodiment 11 0.940 0.557 1.498 Example 12 0.922 0.541 1.463 Example 13 0.935 0.555 1.490 Embodiment 14 0.949 0.539 1.488 Embodiment 15 0.937 0.543 1.480 Example 16 0.931 0.542 1.473 Comparative Example 5 0.855 0.458 1.313 Comparative Example 6 0.824 0.417 1.241 Comparative Example 7 0.906 0.489 1.396 Comparative Example 8 0.788 0.370 1.158

[0220] As shown in Table 3, the total amount of arginine monoglycoside, arginine disaccharide and arginine glycoside in Examples 9-16 is higher than that in Comparative Examples 5-8, indicating that the synergistic fermentation of Bacillus licheniformis and Bacillus subtilis can metabolize and decompose the protein in American ginseng to produce arginine, and degrade starch, cellulose, etc. to produce glucose and maltose, providing a good substrate basis for the synthesis of arginine glycosides, and increasing the content of arginine glycosides in American ginseng extracts. At the same time, the addition of catechins is beneficial to prevent the excessive generation of end products by the Maillard reaction, or the Maillard reaction of other amino-containing substances and carbonyl substances in subsequent steps to produce by-products, thereby reducing the quality of American ginseng extracts. In addition, the enzyme system produced by the synergistic fermentation of Bacillus licheniformis and Bacillus subtilis is more diverse and more active than that of a single strain or exogenously added enzymes. The composite strains have a mutual promotion effect, which can complete the American ginseng fermentation process better and faster, and further increase the content of arginine glycosides.

[0221] The above is only a preferred embodiment of the present invention and does not limit the present invention in any form. Any technician familiar with the art can make many possible changes and modifications to the technical solution of the present invention by using the above disclosed methods and technical contents without departing from the spirit and technical solution of the present invention, or modify it into an equivalent embodiment of equivalent changes. Therefore, any simple modification, equivalent replacement, 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 falls within the scope of protection of the technical solution of the present invention.

Claims

1. A method for increasing arginine glycoside in red ginseng extract or American ginseng extract, characterized in that: The following steps are involved: S1. Select good quality red ginseng or American ginseng, crush and pass through a 50-80 mesh sieve to obtain red ginseng powder or American ginseng powder; S2. Add water to red ginseng powder or American ginseng powder at a solid-liquid ratio of 1: (2-5) and stir to mix evenly to prepare a red ginseng fermentation matrix or an American ginseng fermentation matrix; S3. Inoculate 2-8% of the composite bacterial solution in the red ginseng fermentation matrix or the American ginseng fermentation matrix, and ferment at 30-40 ° C for 12-48h to obtain a red ginseng fermentation liquid or an American ginseng fermentation liquid; S4. The red ginseng fermentation liquid or the American ginseng fermentation liquid is heated at 50-100° C. for 1-3 hours, and then ethanol is added at a solid-liquid ratio of 1:(2-5) and refluxed for extraction for 1-3 hours to obtain an alcohol-extracted red ginseng fermentation liquid or an alcohol-extracted American ginseng fermentation liquid; S5. alcohol-extracted red ginseng fermentation liquid or alcohol-extracted American ginseng fermentation liquid is mixed with water at a solid-liquid ratio of 1: (5-10), and then 0.01-0.04% catechins are added, and the mixture is subjected to heat reflux extraction for 1-3 hours, and concentrated under reduced pressure to obtain red ginseng extract or American ginseng extract; In step S3, the composite bacterial species are Bacillus licheniformis and Bacillus subtilis, and the ratio of Bacillus licheniformis to Bacillus subtilis is (3-1): (1-3).

2. The method for increasing arginine glycoside in red ginseng extract or American ginseng extract according to claim 1, characterized in that: The heating temperature of the ethanol reflux extraction in step S4 is 60-80° C., and the number of extractions is 1-3 times.

3. The method for increasing arginine glycoside in red ginseng extract or American ginseng extract according to claim 1, characterized in that: The heating temperature of the hot reflux extraction in step S5 is 60-100° C., and the number of extractions is 1-3 times.

4. The method for increasing arginine glycoside in red ginseng extract or American ginseng extract according to claim 1, characterized in that: The activation method of Bacillus licheniformis comprises the following steps: Step 1. Thaw the frozen Bacillus licheniformis and inoculate it into the sterilized LB solid medium, and culture it overnight at 28-32°C until colonies grow; Step 2. Pick a single colony of Bacillus licheniformis with an inoculation loop and inoculate it into a sterilized LB liquid culture medium, and culture it at 28-32° C. and 150-200 rpm for 12-15 hours to obtain a Bacillus licheniformis seed solution; Step 3. Inoculate 1-5% of Bacillus licheniformis seed solution into the fermentation medium. The initial strain concentration of the fermentation medium is (2-4)×10 5 CFU / mL, and cultured at 28-32℃, 150-200rpm for 24h to obtain activated Bacillus licheniformis bacterial solution, the concentration of which was (5-8)×10 8 CFU / mL.

5. The method for increasing arginine glycoside in red ginseng extract or American ginseng extract according to claim 1, characterized in that: The activation method of Bacillus subtilis comprises the following steps: Step 4. Thaw the frozen Bacillus subtilis and inoculate it into the sterilized LB solid medium, and culture it overnight at 32-37°C until colonies grow; Step 5. Pick a single colony of Bacillus subtilis with an inoculation loop and inoculate it into a sterilized LB liquid culture medium, and culture it at 32-37° C. and 120-180 rpm for 12-15 hours to obtain a Bacillus subtilis seed solution; Step 6. Inoculate 1-5% Bacillus subtilis seed solution in the fermentation medium. The initial strain concentration of the fermentation medium is (3-4)×10 5 CFU / mL, and cultured at 32-37℃, 150-200rpm for 24h to obtain activated Bacillus subtilis culture solution with a concentration of (7-9)×10 8 CFU / mL.