High-activity ginseng fermented product and application thereof

Abstract

The invention discloses a high-activity ginseng fermented product and application thereof, a compound strain composed of autonomously screened lactobacillus plantarum Byf-r-01 and hansenula polymorpha Byf-j-01 is adopted to ferment ginseng, metabolic enzyme systems of the compound strain are highly complementary, prototype ginsenoside is synergistically and efficiently converted into rare ginsenoside, and the high-activity ginseng fermented product is prepared. The contents of polyphenol, flavone, alkaloid and volatile oil are synchronously increased. According to the invention, the chitosamine-polyaspartic acid compound is added to neutralize lactic acid to maintain the pH stable, and is used as a nitrogen source to promote the proliferation of composite strains, so that the biomass of the strains and the activity of saponin-related invertase are remarkably improved, and the conversion of ginsenoside is promoted. The high-activity ginseng fermentation product prepared by the invention has strong anti-oxidation and anti-fatigue functions, animal experiments prove that the exhaustive swimming time of mice can be prolonged, the blood urea nitrogen level can be reduced, the liver / muscle glycogen reserve can be increased, lactic acid accumulation after exercise can be remarkably relieved, and a new path is provided for development of ginseng high value-added functional products.

Description

Technical Field

[0001] The present invention belongs to the technical field of ginseng fermentation, and specifically relates to a highly active ginseng fermentation product and its application. Background Art

[0002] Ginseng, also known as the "king of all herbs", is a precious medicinal material highly regarded in traditional Chinese medicine, with a long history and extensive medicinal value. It contains rich active polysaccharides, ginsenosides, alkaloids, volatile oils, etc., and has functions such as antioxidant, immune enhancement, anti-fatigue, anti-aging, blood sugar and blood pressure regulation.

[0003] Ginsenosides are the main active components of ginseng. They are divided into prototype ginsenosides (such as Rb1, Rd, Re, etc.) and rare ginsenosides (such as Rg3, Rh1, Rh2, etc.). The content of prototype ginsenosides accounts for about 80% of the total ginsenoside content, but the physiological activity of prototype ginsenosides is low and it is difficult to be absorbed and utilized by the human body; compared with prototype ginsenosides, rare ginsenosides have better physiological functions and are more easily absorbed by the body, but the content of rare ginsenosides in ginseng is very low. Improving the content of rare ginsenosides and realizing the enrichment and extraction of the effective components of ginseng has always been a research hotspot. Therefore, the conversion of prototype ginsenosides in ginseng and the improvement of other bioactive components can not only enhance the utility value of ginseng products, but also broaden the application direction of the ginseng industry. Summary of the Invention

[0004] Technical problems to be solved: Aiming at the above technical problems, the purpose of the present invention is a highly active ginseng fermentation product and its application. The ginseng is fermented with a composite strain composed of the self-screened Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01. The metabolic enzyme systems of the composite strains are highly complementary, and they synergistically and efficiently convert prototype ginsenosides into rare ginsenosides, while simultaneously increasing the contents of polyphenols, flavonoids, alkaloids and volatile oils. The present invention adds a chitosan-polyaspartic acid complex to neutralize lactic acid to maintain pH stability, and at the same time promotes the proliferation of the composite strains as a nitrogen source, significantly improving the biomass of the strains and the activity of saponin-related converting enzymes, and promoting the conversion of ginsenosides. The highly active ginseng fermentation product prepared by the present invention has both strong antioxidant and anti-fatigue functions. Animal experiments have confirmed that it can extend the exhaustive swimming time of mice, reduce the blood urea nitrogen level, increase the liver / muscle glycogen reserve, and significantly relieve the accumulation of lactic acid after exercise, providing a new path for the development of high-value functional products of ginseng.

[0005] Technical solution: A highly active ginseng fermentation product, which is prepared by fermenting ginseng with a composite strain composed of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01; the Lactobacillus plantarum ( Lactobacillus plantarum)Byf-r-01 was deposited at the Guangdong Provincial Culture Collection of Microorganisms on January 20, 2025, with the deposit number GDMCC NO: 65834; the Hanseniaspora uvarum Hanseniaspora uvarum )Byf-j-01 was deposited at the Guangdong Provincial Culture Collection of Microorganisms on January 20, 2025, with the deposit number GDMCC NO: 65833.

[0006] A preparation method of a highly active ginseng fermentation product, comprising the following steps: S1. Select ginseng with good appearance (no disease spots or mildew spots), ultrafinely crush it to a particle size of 0.5 - 1 μm to obtain ginseng powder, add water and mix to form a mixed solution, and perform ultrasonic treatment to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 to prepare Lactobacillus plantarum Byf-r-01 bacterial liquid, and activate Hanseniaspora uvarum Byf-j-01 to prepare Hanseniaspora uvarum Byf-j-01 bacterial liquid; S3. Mix the Lactobacillus plantarum Byf-r-01 bacterial liquid and the Hanseniaspora uvarum Byf-j-01 bacterial liquid to form a compound bacterial liquid, add the compound bacterial liquid and chitosan-polyaspartic acid complex to the ginseng homogenate and ferment, filter and sterilize, and freeze-dry to obtain a highly active ginseng fermentation product.

[0007] Furthermore, in S1, the ratio of ginseng powder to water is 1 g: (4.5 - 8) mL; the conditions for ultrasonic treatment are ultrasonic power of 200 - 600 W and ultrasonic time of 10 - 20 min.

[0008] Furthermore, the preparation method of the Lactobacillus plantarum Byf-r-01 bacterial liquid in S2: Inoculate Lactobacillus plantarum Byf-r-01 into MRS liquid medium at an inoculation amount of 2 - 5%, and statically culture at 30 - 37 °C for 12 - 15 h to obtain Lactobacillus plantarum Byf-r-01 seed liquid; inoculate 1 - 5% of the Lactobacillus plantarum Byf-r-01 seed liquid into MRS liquid medium, and expand the culture at 30 - 37 °C and 150 - 200 rpm for 20 - 24 h, centrifuge and resuspend with sterile water to obtain Lactobacillus plantarum Byf-r-01 bacterial liquid.

[0009] Further, the preparation method of the Hanseniaspora uvarum Byf-j-01 bacterial liquid in S2: inoculate Hanseniaspora uvarum Byf-j-01 at an inoculation amount of 1.5-5% into YPD liquid medium, and statically culture at 28-32°C for 12-15 h to obtain the Hanseniaspora uvarum Byf-j-01 seed liquid; inoculate 1-5% of the Hanseniaspora uvarum Byf-j-01 seed liquid into YPD liquid medium, and perform enlarged culture at 28-32°C and 150-200 rpm for 20-24 h, and centrifuge and resuspend with sterile water to obtain the Hanseniaspora uvarum Byf-j-01 bacterial liquid.

[0010] Further, the volume ratio of the Lactobacillus plantarum Byf-r-01 bacterial liquid to the Hanseniaspora uvarum Byf-j-01 bacterial liquid in S3 is (1.5-5):(1-3.5); the addition amount of the compound bacterial liquid is 2.5-6 vt%; the bacterial concentration of the compound bacterial strain is (1.5-7)×10 8 CFU / mL.

[0011] Further, the preparation method of the chitosan-polyaspartic acid complex in S3: mix chitosan and polyaspartic acid evenly according to the mass ratio of (1-4):1 and dissolve in water, and treat at 20-30°C for 1-3 h, and obtain the chitosan-polyaspartic acid complex after drying.

[0012] Further, the addition amount of the chitosan-polyaspartic acid complex in S3 is 2.5-5 wt%.

[0013] Further, the specific conditions for fermentation in S3 are fermentation temperature 30-35°C and fermentation time 1-4 days; the specific operation of freeze-drying: place the filtered and sterilized ginseng fermentation broth in a vacuum freeze-dryer for freeze-drying, and the specific conditions for freeze-drying are pre-freezing temperature -40 - -30°C, pre-freezing time 1-3 h, heating temperature 10-15°C, drying duration 12-24 h, and vacuum degree 10-20 kPa.

[0014] Application of the above-mentioned high-activity ginseng fermentation product in the preparation of anti-fatigue drugs.

[0015] Further, the forms of the drug include tablets, capsules, extracts, and granules.

[0016] Beneficial effects

[0017] 1. The present invention uses a composite strain composed of self-screened strains (Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01) to ferment ginseng. The biological enzyme system produced by their metabolism has highly complementary characteristics, which can convert the prototype ginsenosides contained in ginseng into rare ginsenosides, increasing the content of rare ginsenosides in highly active ginseng fermentation products. At the same time, it increases the contents of polyphenols, flavonoids and alkaloids. In addition, the cellulase produced by the strain metabolism enzymatically hydrolyzes the cellulose in the ginseng cell wall, releasing the bound volatile oil molecules, thereby increasing the volatile oil content, and then significantly increasing the active ingredients of the ginseng fermentation product, obtaining a highly active ginseng fermentation product.

[0018] 2. During the preparation of the highly active ginseng fermentation product of the present invention, a chitosan-polyaspartic acid complex is added. It is prepared by generating an amide bond between the amino group of chitosan and the carboxyl group of polyaspartic acid. It can neutralize the lactic acid produced by the metabolism of Lactobacillus plantarum Byf-r-01, maintaining the pH stability of the fermentation system. And it can be absorbed and utilized by Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 as a nitrogen source, providing nutrients and energy for the fermentation metabolism of the strains, promoting the proliferation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01, significantly increasing their biomass and vitality, indirectly increasing the synthesis amount and activity of the enzyme system related to the conversion of rare ginsenosides, and finally efficiently converting them into various rare ginsenosides.

[0019] 3. The highly active ginseng fermentation product prepared by the present invention has, on the one hand, high antioxidant capacity and Fe 3+ reduction ability, and can significantly scavenge DPPH free radicals, ABTS+ free radicals, hydroxyl free radicals and superoxide anion free radicals; on the other hand, it has good anti-fatigue function, can significantly prolong the exhaustive swimming time of mice, significantly reduce the blood urea nitrogen level of mice after load-bearing swimming, increase the reserves of liver glycogen and muscle glycogen in the body, increase the content of lactate dehydrogenase in the mouse body, and reduce the accumulation of lactic acid after exercise.

[0020] 4. The present invention uses Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 for composite fermentation to prepare a highly active ginseng fermentation product, which has both high activity and good antioxidant and anti-fatigue functions, and can provide a theoretical basis and data support for the development and application of ginseng functional products. Description of the Drawings

[0021] Figure 1 For the contents of rare ginsenosides in Example 2, Comparative Examples 1-4 and the sample before fermentation; Figure 2 For the free radical scavenging rates of Example 2 and Comparative Examples 1-4, where the free radicals include DPPH•, ABTS+•, •OH, O2 - •; Figure 3 For the exhaustive swimming time, urea nitrogen content, liver glycogen content, and muscle glycogen content of the mice in Example 2 group and Comparative Examples 1-4 group, where A represents the exhaustive swimming time, B represents the urea nitrogen content, C represents the liver glycogen content, and D represents the muscle glycogen content; Figure 4 For the lactate dehydrogenase content of the mice in Example 2 group and Comparative Examples 1-4 group. Detailed implementation method

[0022] The present invention will be further described below in conjunction with embodiments. The following embodiments are explanations of the present invention and the present invention is not limited to the following embodiments: The bacterial strains used in the present invention: ① Lactobacillus plantarum ( Lactobacillus plantarum ) Byf-r-01: Preservation number GDMCC NO: 65834, preservation time is January 20, 2025, and the preservation unit is Guangdong Provincial Microbial Culture Collection Center; ② Hanseniaspora uvarum ( Hanseniaspora uvarum ) Byf-j-01: Preservation number GDMCC NO: 65833, preservation time is January 20, 2025, and the preservation unit is Guangdong Provincial Microbial Culture Collection Center.

[0023] Example 1

[0024] A preparation method of a high-activity ginseng fermentation product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 into 250 mL of MRS liquid medium according to an inoculation amount of 5 mL, and statically culture at 37 °C for 12 h to obtain Lactobacillus plantarum Byf-r-01 seed liquid; inoculate 5 mL of Lactobacillus plantarum Byf-r-01 seed liquid into 250 mL of MRS liquid medium, and perform enlarged culture at 37 °C and 200 rpm for 24 h, centrifuge and resuspend with sterile water to obtain Lactobacillus plantarum Byf-r-01 bacterial liquid; ② Inoculate Hanseniaspora uvarum Byf-j-01 with an inoculum size of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan oligosaccharide and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosan oligosaccharide-polyaspartic acid complex after drying; S4. Mix 30 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (with a bacterial concentration of 4.50×10 8 CFU / mL), add 12 mL of the compound bacterial solution and 9 g of the chitosan oligosaccharide-polyaspartic acid complex to 300 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place in a vacuum freeze dryer for freeze-drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0025] Example 2

[0026] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultrafinely pulverize it to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat it at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 with an inoculum size of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 with an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan amine and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosan amine-polyaspartic acid complex after drying; S4. Mix 30 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the concentration of the bacterial species is 4.55×10 8 CFU / mL), add 10 mL of the compound bacterial solution and 7.5 g of the chitosan amine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0027] Example 3

[0028] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, and obtain ginseng powder by ultrafine pulverization. Add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 with an inoculation amount of 5 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 with an inoculation amount of 10 mL into 250 mL of YPD liquid medium, and culture it statically at 30 °C for 15 h to obtain the Hanseniaspora uvarum Byf-j-01 seed liquid; inoculate 7.5 mL of the Hanseniaspora uvarum Byf-j-01 seed liquid into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain the Hanseniaspora uvarum Byf-j-01 bacterial liquid; S3. Weigh 30 g of chitosanamine and 10 g of polyaspartic acid, mix them evenly and dissolve them in 300 mL of water, treat them at 25 °C for 1.5 h, and dry to obtain the chitosanamine-polyaspartic acid complex; S4. Mix 30 mL of the Lactobacillus plantarum Byf-r-01 bacterial liquid and 20 mL of the Hanseniaspora uvarum Byf-j-01 bacterial liquid to prepare a composite bacterial liquid (the concentration of the bacterial species is 4.50×10 8 CFU / mL), add 12 mL of the composite bacterial liquid and 9 g of the chitosanamine-polyaspartic acid complex to 300 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0029] Example 4

[0030] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, and obtain ginseng powder by ultrafine pulverization. Add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 with an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and culture it statically at 37 °C for 12 h to obtain the Lactobacillus plantarum Byf-r-01 seed liquid; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed liquid into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain the Lactobacillus plantarum Byf-r-01 bacterial liquid; ②Inoculate Hanseniaspora uvarum Byf-j-01 with an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend it with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan amine and 10 g of polyaspartic acid, mix them evenly and dissolve them in 300 mL of water, treat them at 25 °C for 1.5 h, and dry them to obtain a chitosan amine-polyaspartic acid complex; S4. Mix 15 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 35 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the bacterial concentration is 4.65×10 8 CFU / mL). Add 10 mL of the compound bacterial solution and 7.5 g of the chitosan amine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0031] Example 5

[0032] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize it to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat it at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ①Inoculate Lactobacillus plantarum Byf-r-01 with an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend it with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosanamine and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosanamine-polyaspartic acid complex after drying; S4. Mix 40 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 10 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the bacterial species concentration is 4.55×10 8 CFU / mL), add 12 mL of the compound bacterial solution and 9 g of the chitosanamine-polyaspartic acid complex to 300 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze-drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0033] Example 6

[0034] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 at an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculum size of 5 mL into 250 mL of YPD liquid medium, and incubate statically at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h. Centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution. S3. Weigh 30 g of chitosan oligosaccharide and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and dry to obtain a chitosan oligosaccharide-polyaspartic acid complex. S4. Mix 30 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the concentration of the bacterial strain is 4.50×10 8 CFU / mL). Add 10 mL of the compound bacterial solution and 12 g of the chitosan oligosaccharide-polyaspartic acid complex to 400 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0035] Example 7

[0036] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate. S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 at an inoculum size of 10 mL into 250 mL of MRS liquid medium, and incubate statically at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h. Centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution. ②Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend it with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan oligosamine and 10 g of polyaspartic acid, mix them evenly and dissolve them in 300 mL of water, treat them at 25 °C for 1.5 h, and dry them to obtain a chitosan oligosamine-polyaspartic acid complex; S4. Mix 30 mL of the Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of the Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the concentration of the bacterial species is 4.51×10 8 CFU / mL), add 15 mL of the compound bacterial solution and 7.5 g of the chitosan oligosamine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0037] Example 8

[0038] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, and obtain ginseng powder by ultrafine pulverization. Add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ①Inoculate Lactobacillus plantarum Byf-r-01 at an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend it with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 with an inoculum size of 5 mL into 250 mL of YPD liquid medium, and culture it statically at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and culture it under expansion at 30 °C and 150 rpm for 24 h. Centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan oligosaccharide and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosan oligosaccharide-polyaspartic acid complex after drying; S4. Mix 30 mL of the Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of the Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the concentration of the bacterial strain is 4.50×10 8 CFU / mL). Add 12 mL of the compound bacterial solution and 7.5 g of the chitosan oligosaccharide-polyaspartic acid complex to 300 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0039] Example 9

[0040] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 with an inoculum size of 10 mL into 250 mL of MRS liquid medium, and culture it statically at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and culture it under expansion at 37 °C and 200 rpm for 24 h. Centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ②Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan amine and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosan amine-polyaspartic acid complex after drying; S4. Mix 30 mL of the Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of the Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the concentration of the bacterial strain is 4.53×10 8 CFU / mL), add 10 mL of the compound bacterial solution and 7.5 g of the chitosan amine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 35 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0041] Example 10

[0042] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize it to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat it at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ①Inoculate Lactobacillus plantarum Byf-r-01 at an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculation amount of 5 mL into 250 mL of YPD liquid medium, and statically culture it at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and expand the culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosanamine and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosanamine-polyaspartic acid complex after drying; S4. Mix 30 mL of the Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of the Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (with a bacterial concentration of 4.50×10 8 CFU / mL), add 10 mL of the compound bacterial solution and 7.5 g of the chitosanamine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 1.5 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0043] Comparative Example 1

[0044] The difference between this comparative example and Example 2 is that the chitosanamine-polyaspartic acid complex is not added.

[0045] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultra-finely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01 ① Inoculate Lactobacillus plantarum Byf-r-01 at an inoculation amount of 10 mL into 250 mL of MRS liquid medium, and statically culture it at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; ② Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculum size of 5 mL into 250 mL of YPD liquid medium, and incubate statically at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and perform enlarged culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Mix 30 mL of Lactobacillus plantarum Byf-r-01 bacterial solution and 20 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution to prepare a compound bacterial solution (the bacterial strain concentration is 4.55×10 8 CFU / mL), add 10 mL of the compound bacterial solution to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place it in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a high-activity ginseng fermented product.

[0046] Comparative Example 2

[0047] The difference between this comparative example and Example 2 is that only Hanseniaspora uvarum Byf-j-01 is used.

[0048] A preparation method of a high-activity ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, and obtain ginseng powder by ultrafine pulverization. Add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment at an ultrasonic power of 450 W for 15 min to obtain ginseng homogenate; S2. Activation of Hanseniaspora uvarum Byf-j-01 ① Inoculate Hanseniaspora uvarum Byf-j-01 at an inoculum size of 5 mL into 250 mL of YPD liquid medium, and incubate statically at 30 °C for 15 h to obtain a Hanseniaspora uvarum Byf-j-01 seed solution; inoculate 5 mL of the Hanseniaspora uvarum Byf-j-01 seed solution into 250 mL of YPD liquid medium, and perform enlarged culture at 30 °C and 150 rpm for 24 h, then centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum Byf-j-01 bacterial solution; S3. Weigh 30 g of chitosan oligosaccharide and 10 g of polyaspartic acid, mix them evenly and dissolve them in 300 mL of water, treat them at 25 °C for 1.5 h, and obtain a chitosan oligosaccharide-polyaspartic acid complex after drying; S4. Add 10 mL of Hanseniaspora uvarum Byf-j-01 bacterial solution (the bacterial strain concentration is 4.50×10 8CFU / mL) and 7.5 g of chitosan oligosaccharide-polyaspartic acid complex, fermented at 32 °C for 3 days, filtered and sterilized, and freeze-dried in a vacuum freeze dryer (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a highly active ginseng fermented product.

[0049] Comparative Example 3

[0050] The difference between this comparative example and Example 2 is that only Lactobacillus plantarum Byf-r-01 is used.

[0051] A preparation method of a highly active ginseng fermented product, comprising the following steps: S1. Select ginseng with good appearance, ultrafinely pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and ultrasonically treat for 15 min at an ultrasonic power of 450 W to obtain ginseng homogenate; S2. Activation of Lactobacillus plantarum Byf-r-01 ① Inoculate Lactobacillus plantarum Byf-r-01 into 250 mL of MRS liquid medium according to an inoculation amount of 10 mL, and statically culture at 37 °C for 12 h to obtain a Lactobacillus plantarum Byf-r-01 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum Byf-r-01 seed solution into 250 mL of MRS liquid medium, and expand the culture at 37 °C and 200 rpm for 24 h, centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum Byf-r-01 bacterial solution; S3. Weigh 30 g of chitosan oligosaccharide and 10 g of polyaspartic acid, mix evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosan oligosaccharide-polyaspartic acid complex after drying; S4. Add 10 mL of Lactobacillus plantarum Byf-r-01 bacterial solution (its bacterial concentration is 4.50×10 8 CFU / mL) and 7.5 g of chitosan oligosaccharide-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and freeze-dry in a vacuum freeze dryer (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a highly active ginseng fermented product.

[0052] Comparative Example 4

[0053] The difference between this comparative example and Example 2 is that commercially available Lactobacillus plantarum and Hanseniaspora uvarum are used, and Lactobacillus plantarum M32-2 (CICC20314) and Hanseniaspora uvarum F30 (CICC 31872) are both purchased from the China Center for Industrial Culture Collection.

[0054] A preparation method of a highly active ginseng fermentation product, comprising the following steps: S1. Select ginseng with good appearance, ultramicro pulverize to obtain ginseng powder, add 500 mL of water to 100 g of ginseng powder and mix to form a mixed solution, and perform ultrasonic treatment for 15 min at an ultrasonic power of 450 W to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum M32-2 and Hanseniaspora uvarum F30 ① Inoculate Lactobacillus plantarum M32-2 into 250 mL of MRS liquid medium according to an inoculation amount of 10 mL, and statically culture at 37 °C for 12 h to obtain a Lactobacillus plantarum M32-2 seed solution; inoculate 7.5 mL of the Lactobacillus plantarum M32-2 seed solution into 250 mL of MRS liquid medium, and perform enlarged culture at 37 °C and 200 rpm for 24 h, centrifuge and resuspend with sterile water to obtain a Lactobacillus plantarum M32-2 bacterial solution; ② Inoculate Hanseniaspora uvarum F30 into 250 mL of YPD liquid medium according to an inoculation amount of 5 mL, and statically culture at 30 °C for 15 h to obtain a Hanseniaspora uvarum F30 seed solution; inoculate 5 mL of the Hanseniaspora uvarum F30 seed solution into 250 mL of YPD liquid medium, and perform enlarged culture at 30 °C and 150 rpm for 24 h, centrifuge and resuspend with sterile water to obtain a Hanseniaspora uvarum F30 bacterial solution; S3. Weigh 30 g of chitosanamine and 10 g of polyaspartic acid, mix them evenly and dissolve in 300 mL of water, treat at 25 °C for 1.5 h, and obtain a chitosanamine-polyaspartic acid complex after drying; S4. Mix 30 mL of the Lactobacillus plantarum M32-2 bacterial solution and 20 mL of the Hanseniaspora uvarum F30 bacterial solution to form a composite bacterial solution (the bacterial species concentration is 4.55×10 8 CFU / mL), add 10 mL of the composite bacterial solution and 7.5 g of the chitosanamine-polyaspartic acid complex to 250 mL of ginseng homogenate, ferment at 32 °C for 3 days, filter and sterilize, and place in a vacuum freeze dryer for freeze drying (pre-freezing temperature -35 °C, pre-freezing time 2.5 h, heating temperature 15 °C, drying duration 24 h, vacuum degree 18 kPa) to obtain a highly active ginseng fermentation product.

[0055] Performance test

[0056] (1) Polysaccharide content Using glucose as a standard product, the polysaccharide content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined by the phenol-sulfuric acid method.

[0057] (2) Total phenol content Using gallic acid as a standard, the total phenolic content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined by the Folin-Ciocalteu colorimetric method.

[0058] (3)Total flavonoid content Using rutin as a standard, the total flavonoid content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined by the aluminum chloride colorimetric method.

[0059] (4)Total saponin content Using ginsenoside Re as a standard, the total saponin content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined by the vanillin-glacial acetic acid method.

[0060] (5)Alkaloid content The total alkaloid content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined by the acidic dye colorimetric method.

[0061] (6)Volatile oil content According to the determination method of volatile oil in the fourth part of the Chinese Pharmacopoeia 2020 Edition (2204), the volatile oil content in the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4 was determined.

[0062] Table 1 Physicochemical indexes of the highly active ginseng fermentation products prepared in Examples 1-10 and Comparative Examples 1-4

[0063] As can be seen from Table 1, the polysaccharide and total saponin contents of Examples 1-10 and Comparative Examples 1-4 were lower than those of the samples before fermentation. This is because Lactobacillus plantarum and Hanseniaspora uvarum need polysaccharides in ginseng as a carbon source for growth and metabolism, and the rich enzyme systems produced by metabolism convert protopanaxadiol-type ginsenosides into rare ginsenosides, so the polysaccharide and total saponin contents decreased; while the total phenols, total flavonoids, alkaloids and volatile oils of Examples 1-10 were higher than those of the samples before fermentation and also higher than those of Comparative Examples 1-4. This shows that Lactobacillus plantarum Byf-r-1 and Hanseniaspora uvarum Byf-j-1 used in the examples can efficiently metabolize macromolecular substances in ginseng into polyphenols and flavonoids. At the same time, chitosan-polyaspartic acid complex can not only serve as a nitrogen source to supply energy for the strains, but also neutralize the lactic acid produced by Lactobacillus plantarum Byf-r-1 to maintain the pH stability of the fermentation system, improve the activity of related enzymes, promote the production of alkaloids and volatile oils in the highly active ginseng fermentation products, thereby increasing the bioactive substances in the highly active ginseng fermentation products and endowing them with excellent biological effects.

[0064] (7)Quantitative analysis of rare ginsenosides Analyze the rare ginsenosides in the highly active ginseng fermentation products prepared in Example 2 and Comparative Examples 1-4 according to "GB / T 41726-2022 Identification and Detection Methods of Ginsenoside Monomers".

[0065] It can be seen from Figure 1 that the rare ginsenosides Rd, Rg3, Rh1, Rh2 and Rh4 in the highly active ginseng fermentation products prepared in Example 2 and Comparative Examples 1-4 are all at relatively high levels, and the rare ginsenosides in Example 2 are all higher than those in Comparative Examples 1-4. This shows that on the one hand, the combined use of self-screened bacteria (Lactobacillus plantarum Byf-r-1 and Hanseniaspora uvarum Byf-j-1) has highly complementary characteristics in the metabolic biological enzyme systems, and can efficiently decompose the protopanaxadiol ginsenosides in ginseng into rare ginsenosides; on the other hand, the addition of chitosan-polyaspartic acid complex can not only provide the nutrients and energy required by the complex strains as a nitrogen source, but also improve the activity of the enzyme systems related to the conversion of protopanaxadiol ginsenosides into rare ginsenosides, thereby increasing the production of rare ginsenosides.

[0066] Example 11

[0067] Verification test on the antioxidant capacity of highly active ginseng fermentation products Measure the DPPH free radical scavenging ability, ABTS cation free radical scavenging ability, hydroxyl free radical scavenging ability, superoxide anion free radical scavenging ability and Fe 3+ reducing power of the highly active ginseng fermentation products prepared in Example 2 and Comparative Examples 1-4 respectively, as follows: (1) DPPH free radical scavenging ability: Prepare a sample solution with a mass concentration of 500 μg / mL and a positive control V C solution. Add 20 μL of the sample and positive control solution and 180 μL of the DPPH working solution into a 96-well plate, place it in an incubator at 37 °C for light-shielded reaction for 10 min, measure the absorbance at 517 nm. The blank group is absolute ethanol + DPPH working solution, and the background group is the sample / positive control Vc solution + absolute ethanol. Calculate the DPPH free radical scavenging rate according to the formula; the formula is: DPPH free radical scavenging rate (%) = [1 - (A1 - A2) / A0] × 100%, where A1 is the absorbance of the sample / positive control group, A2 is the absorbance of the background group of the positive control / sample, and A0 is the absorbance of the blank group; (2)ABTS cation radical scavenging ability: Prepare a sample solution at 500 μg / mL and a positive control Vc solution. Add 20 μL of the sample and positive control solutions and 180 μL of the ABTS working solution to a 96-well plate. Set three parallels for each concentration. Place it in an incubator at 37°C and react in the dark for 10 min. Measure the absorbance of each group at 734 nm; the blank group is distilled water + ABTS working solution, and the background group is the sample / positive control Vc solution + distilled water; calculate the ABTS+ radical scavenging rate according to the formula; the formula is: ABTS+ radical scavenging rate (%) = [1 - (A1 - A2) / A0] × 100%, where A1 is the absorbance of the sample / positive control group, A2 is the absorbance of the background group of the positive control / sample, and A0 is the absorbance of the blank group; (3)Hydroxyl radical scavenging ability: Take 1 mL each of 1.8 mmol / L FeSO4, 1.8 mmol / L salicylic acid-ethanol solution, and the sample solution and mix well. Then add 1 mL of 1.76 mmol / L H2O2. React in a water bath at 37°C for 30 min. Measure the absorbance value A1 at 510 nm using a microplate reader; take 1 mL of the sample solution and mix it with 3 mL of ethanol, and measure the absorbance value A0 at 510 nm using a microplate reader; take 1 mL each of 1.8 mmol / L FeSO4, 1.8 mmol / L salicylic acid-ethanol solution, and ethanol, then add 1 mL of 1.76 mmol / L H2O2. React in a water bath at 37°C for 30 min. Measure the absorbance value A2 of the solution system without antioxidant at 510 nm using a microplate reader. Calculate the hydroxyl radical scavenging rate according to the formula; the formula is: Hydroxyl radical scavenging rate (%) = [1 - (A1 - A0) / A2] × 100%; (4)Superoxide anion radical scavenging ability: Prepare a sample solution at 1000 μg / mL and a positive control Vc solution. Add 20 μL of the sample solution, 90 μL of buffer solution, 60 μL of distilled water, and 10 μL of pyrogallol solution to a 96-well plate. Set three parallels for each concentration. Place it in an incubator at 25°C and react in the dark for 20 min. Measure the absorbance of each group at 320 nm; the blank group uses 20 μL of distilled water and the subsequent reaction working solution, and the background group is the sample / positive control Vc solution + distilled water. Calculate the superoxide anion radical scavenging rate according to the formula; the formula is: Superoxide anion radical scavenging rate (%) = [1 - (A1 - A2) / A0] × 100%, where A1 is the absorbance of the sample / positive control group, A2 is the absorbance of the background group of the positive control / sample, and A0 is the absorbance of the blank group; (5)Total reducing ability: Prepare a sample solution with a concentration of 500 μg / mL and a positive control Vc solution. Take 300 μL of the sample into a 2 mL centrifuge tube, and then successively add 300 μL of phosphate buffer and 300 μL of potassium ferricyanide solution. After gently shaking and mixing, place it in an incubator at 50 °C for reaction for 20 min. Then immediately cool it in an ice-water bath, and add 300 μL of trichloroacetic acid solution to terminate the reaction. Subsequently, centrifuge each group at 2500 rpm for 5 min. Take 90 μL of the supernatant into a 96-well plate, and successively add 90 μL of distilled water and 18 μL of ferric chloride solution. Let it stand for 10 min, and measure the absorbance value of each group at 700 nm; for the blank control group, use 300 μL of distilled water plus the subsequent reaction solution, and the positive control uses ascorbic acid (Vc); calculate the total reducing power according to the formula, and the formula is total reducing power = A1 - A0, where A1 is the absorbance of the sample / positive control group, and A0 is the absorbance of the blank group.

[0068] Table 2 Total reducing ability of positive control, Example 2, and Comparative Examples 1-4

[0069] From Figure 2 and Table 2, it can be seen that the highly active ginseng fermented product prepared in Example 2 has good scavenging ability for various free radicals (including DPPH•, ABTS+•, •OH, O2 - •), which is better than that of Comparative Examples 1-4, and its total reducing power is also higher than that of Comparative Examples 1-4. Because compared with Comparative Examples 1-4, the highly active ginseng fermented product prepared in Example 2 contains more polyphenols and flavonoids, as well as highly active rare ginsenosides, so its antioxidant function is better than that of Comparative Examples 1-4.

[0070] Example 12

[0071] Verification test on the anti-fatigue function of highly active ginseng fermented product Animal experiments were conducted to analyze the anti-fatigue effects of the highly active ginseng fermented products prepared in Example 2 and Comparative Examples 1-4. Specifically, 70 SPF-grade Kunming mice (male, body weight 18-22 g) were randomly divided into 7 groups (blank control group, positive group, Example 2 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, Comparative Example 4 group), with 10 mice in each group. They were adaptively fed for 3 days. Administration method: ① The blank control group was given an equal volume of distilled water and continuously gavaged for 30 days; ② The positive group was given American ginseng lozenges at a dosage of 0.48 g / kg / d; ③ The Example 2 group, Comparative Example 1 group, Comparative Example 2 group, Comparative Example 3 group, and Comparative Example 4 group were all given the freeze-dried powder of the highly active ginseng fermented product at a dosage of 1.95 g / kg / d. Feeding method: Each group of mice was placed in a standard experimental animal room for feeding. The environmental temperature was 25±1°C, the relative humidity was 50-60%, with day and night alternation. Sufficient sterilized mouse feed and sterilized water were provided. During the feeding period, the feed and water were freely available, and the feeding cycle was 30 days. After the feeding cycle ended, the body weights of each group of mice were weighed, and the mice were subjected to a load-bearing swimming experiment to record the exhaustive swimming time of the mice. Then the mice were sacrificed and sampled to analyze the contents of urea nitrogen, liver glycogen, muscle glycogen, and lactate dehydrogenase.

[0072] Table 3 Initial and final body weights of mice in each treatment group

[0073] As can be seen from Table 3, compared with the blank control group and the positive group, feeding the highly active ginseng fermented products prepared in Example 2 and Comparative Examples 1-4 to mice had little effect on the body weight gain of the mice.

[0074] Fatigue is mainly manifested in the inability to concentrate and a decrease in exercise endurance. The exhaustive exercise duration can be used as an effective means to evaluate the strength of exercise ability. As Figure 3 shown in A, compared with the positive group, the exhaustive swimming time of the mice in the Example 2 group and Comparative Examples 1-3 groups was longer, while that of the mice in the Comparative Example 4 group was shorter; as Figure 3 shown in B, Figure 3 shown in C, and Figure 3 shown in D, the urea nitrogen content of the mice in the Example 2 group was lower than that of the positive group and Comparative Examples 1-4 groups, while the liver glycogen content and muscle glycogen content were higher than those of the positive group and Comparative Examples 1-4 groups; as Figure 4 shown, the lactate dehydrogenase content of the Example 2 group was higher than that of the positive group and Comparative Examples 1-4 groups. Thus, it can be seen that the highly active ginseng fermented product prepared in the Example 2 group has good anti-fatigue ability, can promote the glycogen reserve of the body, reduce the level of urea nitrogen produced after the body's protein metabolism, and the accumulation of lactic acid in the body.

[0075] The above are only the preferred embodiments of the present invention, and do not impose any formal restrictions on the present invention. Any person skilled in the art can make many possible changes and modifications to the technical solution of the present invention by using the methods and technical contents disclosed above, or modify it into equivalent embodiments with equivalent changes, without departing from the spirit and technical solution of the present invention. Therefore, any simple modification, equivalent replacement, equivalent change and modification made to the above embodiments based on the technical essence of the present invention without departing from the technical solution of the present invention still fall within the scope of protection of the technical solution of the present invention.

Claims

1. A highly active fermented ginseng product, characterized in that, The highly active ginseng fermented product is prepared by fermenting ginseng with a composite strain composed of Lactobacillus plantarum Byf-r-01 and Hanseniaspora uvarum Byf-j-01; Lactobacillus plantarum Byf-r-01 was deposited in the Guangdong Provincial Culture Collection of Microorganisms on January 20, 2025, with the deposit number GDMCC NO: 65834; Hanseniaspora uvarum Byf-j-01 was deposited in the Guangdong Provincial Culture Collection of Microorganisms on January 20, 2025, with the deposit number GDMCC NO: 65833.

2. The preparation method of a highly active ginseng fermented product according to claim 1, characterized in that, It includes the following steps: S1. Select ginseng with good appearance, ultramicro pulverize it to a particle size of 0.5 - 1 μm to obtain ginseng powder, add water and mix to form a mixed solution, and perform ultrasonic treatment to obtain ginseng homogenate; S2. Activate Lactobacillus plantarum Byf-r-01 to prepare Lactobacillus plantarum Byf-r-01 bacterial liquid, and activate Hanseniaspora uvarum Byf-j-01 to prepare Hanseniaspora uvarum Byf-j-01 bacterial liquid; S3. Mix the Lactobacillus plantarum Byf-r-01 bacterial liquid and the Hanseniaspora uvarum Byf-j-01 bacterial liquid to form a composite bacterial liquid, add the composite bacterial liquid and chitosan - polyaspartic acid complex to the ginseng homogenate and ferment, filter and sterilize, and freeze-dry to obtain the highly active ginseng fermented product.

3. The preparation method of a highly active ginseng fermented product according to claim 2, wherein: In the above S1, the ratio of ginseng powder to water is 1 g : (4.5 - 8) mL; the conditions for ultrasonic treatment are ultrasonic power of 200 - 600 W and ultrasonic time of 10 - 20 min.

4. The preparation method of a highly active ginseng fermentation product according to claim 2, characterized in that, The volume ratio of the Lactobacillus plantarum Byf-r-01 bacterial liquid to the Hanseniaspora uvarum Byf-j-01 bacterial liquid in S3 is (1.5 - 5):(1 - 3.5); the addition amount of the composite bacterial liquid is 2.5 - 6 vt%; the bacterial concentration of the composite bacterial strain is (1.5 - 7)×10 8 CFU / mL.

5. The preparation method of a highly active ginseng fermented product according to claim 2, characterized in that, The preparation method of the chitosan - polyaspartic acid complex in the above S3: Mix chitosan and polyaspartic acid in a mass ratio of (1 - 4) : 1, dissolve them in water, treat them at 20 - 30 °C for 1 - 3 h, and obtain the chitosan - polyaspartic acid complex after drying.

6. The preparation method of a highly active ginseng fermented product according to claim 2, characterized in that, In the above S3, the addition amount of the chitosan - polyaspartic acid complex is 2.5 - 5 wt%.

7. The preparation method of a highly active ginseng fermented product according to claim 2, characterized in that: The specific conditions for fermentation in the above S3 are fermentation temperature of 30 - 35 °C and fermentation time of 1 - 4 days.

8. The application of the highly active ginseng fermented product according to claim 1 in the preparation of anti-fatigue drugs.

9. The application according to claim 8, wherein The forms of the drug include tablets, capsules, instant granules, and granules.

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