Fermented black ginseng product and preparation method thereof

Abstract

The invention belongs to the technical field of food processing, and particularly discloses a black ginseng fermented product and a preparation method thereof, the method comprises the following steps: step S1, pretreating ginseng to obtain black ginseng powder; s2, preparing a compound fermentation bacteria solution; s3, preparing a culture medium; s4, inoculating and fermenting to obtain a black ginseng fermented concentrated solution; and step S5, carrying out process molding on the black ginseng fermentation concentrated solution and auxiliary materials according to requirements, so as to obtain the black ginseng fermentation product. According to the black ginseng fermented product and the preparation method thereof disclosed by the invention, efficient conversion and enrichment of effective components of black ginseng are realized by optimizing fermentation process parameters, meanwhile, the irritation of the black ginseng fermented product is reduced, the taste is improved, and the prepared black ginseng fermented product is suitable for a wide range of people.

Description

Technical Field

[0001] This invention belongs to the field of food processing technology, specifically relating to a fermented black ginseng product and its preparation method. Background Technology

[0002] The core active ingredient of ginseng (Panax ginseng CA Mey.) has been widely identified as ginsenosides, belonging to the triterpenoid class of compounds (tetracyclic or pentacyclic triterpenoid derivatives). These are the characteristic components that distinguish ginseng from other plants and are the main material basis for its pharmacological effects. Black ginseng, as a deep-processed product of ginseng, is prepared through traditional steaming and drying processes. This traditional process allows some components in ginseng to transform, forming the unique active substances of black ginseng, and is the fundamental step in the preparation of black ginseng products.

[0003] However, relying solely on the traditional preparation process of black ginseng products to produce the final product has obvious drawbacks: on the one hand, the original saponins are difficult for the human body to absorb directly, resulting in low bioavailability; on the other hand, the process cycle is long and energy consumption is high, and the final product has a bitter taste and is somewhat irritating to the stomach and intestines, limiting its consumption by people with sensitive stomachs, the elderly, and other groups.

[0004] To address the aforementioned issues, existing technologies employ microbial fermentation to improve the preparation process of black ginseng products. For example, while fermentation with a single *Lactobacillus plantarum* strain can partially convert the original saponins, the lack of cell wall-degrading enzymes makes it difficult to effectively break down the cell walls of black ginseng, resulting in insufficient saponin release. Furthermore, the single-strain fermentation system has limited functionality, leading to low yields of rare ginseng saponins. Alternatively, a binary system of *Saccharomyces cerevisiae* and lactic acid bacteria can improve saponin conversion efficiency, but it lacks cellulose-degrading bacteria, resulting in incomplete cell wall breakdown. Additionally, the intestinal tolerance of the fermentation products is not effectively improved, and the incidence of discomfort after consumption remains high among individuals with sensitive gastrointestinal tracts.

[0005] Therefore, there is a need in this field to develop a fermented black ginseng product and its preparation method, which can retain the advantages of the traditional black ginseng preparation process, make up for its defects through subsequent process optimization, and thus solve the above problems. Summary of the Invention

[0006] The purpose of this invention is to provide a fermented black ginseng product and its preparation method. By optimizing the fermentation process parameters, the effective components of black ginseng are efficiently converted and enriched, while reducing the irritation of the fermented black ginseng product and improving its taste. As a result, the fermented black ginseng product prepared is suitable for a wide range of people.

[0007] To achieve the above objectives, the present invention provides a method for preparing fermented black ginseng products, comprising the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S3: Prepare the culture medium; Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S5: According to the requirements, the black ginseng fermentation concentrate and excipients are processed into functional black ginseng fermentation products in the form of oral liquid, freeze-dried powder or compressed candy.

[0008] Preferably, step S2 specifically includes: Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Step S22: Mix the bacteria (Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, Cordyceps militaris) in a volume ratio of (2.5~3.2):(1.5~2):(1.8~2.1):(0.5~1). This ratio can achieve complementary metabolic advantages of each strain, avoid antagonistic effects between strains, maximize the conversion of effective ingredients and improve taste, and obtain a compound fermentation liquid. Lactobacillus plantarum produces lactic acid, organic acids and various glycosidases, which promote the deglycosylation reaction of ginsenosides, converting protopanaxadiol-type saponins into rare saponins such as Rh2 and Rg3. At the same time, it lowers the pH value of the product, inhibits the growth of harmful bacteria, and improves intestinal tolerance. Saccharomyces cerevisiae produces protease, amylase and β-glucosidase, which degrade the protein and starch in black ginseng to produce nutrients such as amino acids and small molecule sugars, improving the taste of the product. At the same time, the ethanol produced by its metabolism can promote the dissolution and conversion of ginsenosides. Bacillus subtilis produces cellulase, pectinase and antimicrobial peptides, which disrupt the cell wall structure of black ginseng and promote the release of active ingredients. At the same time, the antimicrobial peptides can enhance the antibacterial properties of the product and extend its shelf life. The B vitamins produced by its metabolism can synergistically enhance the nutritional value of the product. Cordyceps militaris produces cordycepic acid, cordycepin, and various bioactive enzymes, which work synergistically with other microorganisms to further improve the conversion efficiency of rare saponins. At the same time, cordycepic acid can protect the gastrointestinal mucosa, reduce the irritation of fermented black ginseng products, and expand the range of applicable populations.

[0009] Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0010] Preferably, in step S21, the specific conditions for activating and culturing *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* are as follows: Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 18-24 h, viable count ≥5×10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 12-16 h, viable count ≥3×10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 16-20 h, viable count ≥4×10⁻⁶. 9 CFU / mL; Cordyceps militaris: Cultured on PDA medium at 25℃ in the dark for 48-72 hours, spore concentration ≥2×10⁻⁶. 8 CFU / mL.

[0011] Preferably, in step S3, the culture medium comprises the following components by weight percentage: 20-25% black ginseng powder, 3-8% fructooligosaccharides, 0.5-1.5% peptone, 0.1-0.3% KH2PO4, 0.05-0.15% MgSO4, and the remainder is deionized water.

[0012] Preferably, step S3 specifically includes: Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0~6.5; Step S32: Sterilize at 110~120℃ for 20~25 minutes, then cool to room temperature to obtain the culture medium.

[0013] Preferably, step S4 specifically includes: Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 4-7% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; Step S43: Centrifuge, extract, and concentrate the black ginseng fermentation liquid in sequence to obtain black ginseng fermentation concentrate.

[0014] Preferably, the staged gradient fermentation in step S42 specifically involves the following: the temperature of the first stage of aerobic fermentation is 23~27℃, the pH is 6.0~6.5, the fermentation time is 30~40h, and the dissolved oxygen concentration is 5~8mg / L. The second stage of anaerobic fermentation is carried out at a temperature of 30-35℃, a pH of 4.8-5.0, and a fermentation time of 48-72 hours. The third stage of microaerobic fermentation is carried out at a temperature of 30-32℃, a pH of 5.5-6.0, a fermentation time of 30-35 hours, and a dissolved oxygen concentration of 1-3 mg / L.

[0015] Preferably, in step S43, the centrifugation speed is 6000~8000 r / min and the centrifugation time is 10~20 min; Ultrasonic extraction was performed with a power of 200-300W, an extraction temperature of 40-50℃, and an extraction time of 30-45 minutes. The concentration temperature is 50~60℃, the concentration vacuum degree is -0.08~-0.09MPa, and the solid content after concentration is 25~30%.

[0016] Preferably, in step S5, the fermented black ginseng product comprises the following components by weight percentage: 75-80% black ginseng fermentation concentrate and 20-25% excipients; wherein the excipients are one or more of maltitol, fructooligosaccharides, xanthan gum, and citric acid.

[0017] The present invention also provides a method for preparing fermented black ginseng products and the resulting fermented black ginseng products.

[0018] The present invention employs the above-mentioned fermented black ginseng product and its preparation method, and the beneficial effects are as follows: (1) This invention combines plant lactobacillus, brewer's yeast, Bacillus subtilis and Cordyceps militaris to form a compound fermentation strain. Through the synergistic effect of each strain, the efficient conversion of ginsenosides, the degradation of polysaccharides and the improvement of taste are achieved, thereby solving the problem of poor fermentation effect of single strain or conventional compound strain.

[0019] (2) The present invention adopts a three-stage fermentation mode of aerobic-anaerobic-microaerobic, which is adapted to the metabolic characteristics of different strains and significantly increases the content of rare saponins.

[0020] (3) The black ginseng fermented products prepared by the present invention through the synergistic metabolism of compound bacteria and the special design of mild formula have significantly reduced irritation. At the same time, the products have multiple effects such as enhancing immunity and anti-fatigue, and have a suitable sweet and sour taste and rich flavor, thus meeting the needs of different groups of people.

[0021] (4) The preparation process in this invention is simple, the strains used are all commercially available conventional strains, the equipment is conventional equipment in the food processing field, the cost is controllable, and it is easy to carry out large-scale industrial production.

[0022] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0023] Figure 1 This is a flowchart of the preparation process of a fermented black ginseng product and its preparation method according to Example 1 of the present invention. Detailed Implementation

[0024] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0025] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0026] The ginseng used in this invention is produced in Changbai Mountain, Jilin Province; Lactobacillus plantarum CICC 25283, Saccharomyces cerevisiae CICC 32883, Bacillus subtilis CICC 10732, and Cordyceps militaris CICC 14013 were all purchased from the China Industrial Microbial Culture Collection Center. Culture media: MRS medium, YPD medium, LB medium, and PDA medium, all of which are commercially available conventional culture media.

[0027] Example 1 like Figure 1 As shown, a method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder.

[0028] Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0029] Step S22: Mix Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, and Cordyceps militaris in a volume ratio of 3:2:2:1 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0030] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0031] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0032] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 25℃, a pH of 6.5, a fermentation time of 35 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 30℃ and a pH of 5.0 for 60 hours. The third stage of microaerobic fermentation was carried out at a temperature of 30℃, a pH of 5.5, and a fermentation time of 33 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0033] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0034] Step S5: According to the requirements, the black ginseng fermentation concentrate and auxiliary materials are processed into shape to obtain the black ginseng fermentation product.

[0035] By weight percentage, 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water are mixed and stirred, filtered through a 1μm filter membrane, and then bottled and sterilized to obtain black ginseng fermented oral liquid.

[0036] Example 2 A method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0037] Step S22: Mix the following ingredients in a volume ratio of 2.5:1.5:1.8:0.5 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0038] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0039] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0040] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 25℃, a pH of 6.5, a fermentation time of 35 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 30℃ and a pH of 5.0 for 60 hours. The third stage of microaerobic fermentation was carried out at a temperature of 30℃, a pH of 5.5, and a fermentation time of 33 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0041] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0042] Step S5: By weight percentage, mix and stir 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water, filter through a 1μm filter membrane, fill and sterilize to obtain black ginseng fermented oral liquid.

[0043] Example 3 A method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0044] Step S22: Mix Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, and Cordyceps militaris in a volume ratio of 3.2:2:2.1:1 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0045] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0046] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0047] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 25℃, a pH of 6.5, a fermentation time of 35 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 30℃ and a pH of 5.0 for 60 hours. The third stage of microaerobic fermentation was carried out at a temperature of 30℃, a pH of 5.5, and a fermentation time of 33 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0048] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0049] Step S5: By weight percentage, mix and stir 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water, filter through a 1μm filter membrane, fill and sterilize to obtain black ginseng fermented oral liquid.

[0050] Example 4 A method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0051] Step S22: Mix Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, and Cordyceps militaris in a volume ratio of 3:2:2:1 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0052] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0053] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0054] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 27℃, a pH of 6.5, a fermentation time of 35 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 35℃ and a pH of 5.0 for 60 hours. The third stage of microaerobic fermentation was carried out at a temperature of 32℃, a pH of 5.5, and a fermentation time of 33 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0055] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0056] Step S5: By weight percentage, mix and stir 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water, filter through a 1μm filter membrane, fill and sterilize to obtain black ginseng fermented oral liquid.

[0057] Example 5 A method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0058] Step S22: Mix Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, and Cordyceps militaris in a volume ratio of 3:2:2:1 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0059] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0060] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0061] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 25℃, a pH of 6.5, a fermentation time of 30 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 30℃ and a pH of 5.0 for 48 hours. The third stage of microaerobic fermentation was carried out at a temperature of 30℃, a pH of 5.5, and a fermentation time of 30 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0062] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0063] Step S5: By weight percentage, mix and stir 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water, filter through a 1μm filter membrane, fill and sterilize to obtain black ginseng fermented oral liquid.

[0064] Example 6 A method for preparing a fermented black ginseng product includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 20 h, viable count ≥ 5 × 10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 14 h, viable count ≥ 3 × 10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 18 h, viable count ≥ 4 × 10⁻⁶ 9 CFU / mL; Cordyceps militaris: PDA medium, cultured at 25℃ in the dark for 60 h, spore concentration ≥2×10 8 CFU / mL.

[0065] Step S22: Mix Bacillus plantarum, Saccharomyces cerevisiae, Bacillus subtilis, and Cordyceps militaris in a volume ratio of 3:2:2:1 to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

[0066] Step S3: Prepare the culture medium; The culture medium comprises the following components by weight percentage: 25% black ginseng powder, 7% fructooligosaccharides, 1% peptone, 0.2% KH2PO4, 0.1% MgSO4, and the remainder is deionized water.

[0067] Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0; Step S32: Sterilize at 120℃ for 25 minutes, then cool to room temperature to obtain the culture medium.

[0068] Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 5% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; The first stage of aerobic fermentation was carried out at a temperature of 25℃, a pH of 6.5, a fermentation time of 40 hours, and a dissolved oxygen concentration of 6 mg / L. The second stage of anaerobic fermentation was carried out at a temperature of 30℃ and a pH of 5.0 for 72 hours. The third stage of microaerobic fermentation was carried out at a temperature of 30℃, a pH of 5.5, and a fermentation time of 35 hours, with the dissolved oxygen concentration controlled at 2 mg / L.

[0069] Step S43: Centrifuge the black ginseng fermentation broth sequentially at a speed of 7000 r / min for 15 min; extract using ultrasonic-assisted extraction at a power of 250 W, a temperature of 45℃, and a time of 35 min; concentrate at a temperature of 55℃ and a vacuum of -0.08 MPa, resulting in a solid content of 30% to obtain the black ginseng fermentation concentrate.

[0070] Step S5: By weight percentage, mix and stir 80% black ginseng fermentation concentrate, 5% maltitol, 2% fructooligosaccharides, 0.3% citric acid, 0.08% xanthan gum, and 12.62% deionized water, filter through a 1μm filter membrane, fill and sterilize to obtain black ginseng fermented oral liquid.

[0071] Comparative Example 1 The difference from Example 1 is as follows: Step S21, Bacillus plantarum, Saccharomyces cerevisiae, and Cordyceps militaris are activated and cultured separately; Step S22, Bacillus plantarum, Saccharomyces cerevisiae, and Cordyceps militaris are mixed in a volume ratio of 3:2:3 to obtain a compound fermentation broth. The remaining process steps are the same as in Example 1.

[0072] Comparative Example 2 The difference from Example 1 is as follows: Step S21, Bacillus plantarum, Bacillus subtilis, and Cordyceps militaris are activated and cultured separately; Step S22, Bacillus plantarum, Bacillus subtilis, and Cordyceps militaris are mixed in a volume ratio of 3:2:3 to obtain a compound fermentation broth. The remaining process steps are the same as in Example 1.

[0073] Comparative Example 3 The difference from Example 1 is that in step S42, natural fermentation is carried out to obtain black ginseng fermentation liquid; the temperature of natural fermentation is 25°C and the fermentation time is 360 hours. The remaining process steps are the same as in Example 1.

[0074] Experimental Example 1 The performance of the black ginseng fermented oral liquids prepared in Examples 1-6 and Comparative Examples 1-3 was tested.

[0075] (1) Detection of active ingredient content: The contents of ginsenosides Rh2 and Rg3 were determined by high performance liquid chromatography (HPLC), the proportion of small molecule polysaccharides (≤3000Da) was determined by gel permeation chromatography (GPC), and the total amino acid content was determined by an automated amino acid analyzer. The results are shown in Table 1.

[0076] Table 1 Comparison of active ingredient content

[0077] As shown in Table 1, the contents of ginsenosides Rh2 and Rg3 in Examples 1-6 of the present invention are significantly higher than those in Comparative Examples 1-3, the proportion of small molecule polysaccharides is much higher than that in each comparative example, and the total amino acid content is also significantly higher than that in the comparative examples. This indicates that the preparation method of black ginseng fermentation products in the present invention can effectively promote the transformation and enrichment of effective components of black ginseng.

[0078] (2) Detection of immune regulatory function: Phagocytosis experiment of mouse macrophages: Ninety Kunming mice were randomly divided into 9 groups (Examples 1-6, Comparative Examples 1-3), with 10 mice in each group. The dose was 200 mg / kg·d, and the mice were continuously administered by gavage for 30 days. 24 h after the last gavage, the mice were injected intraperitoneally with chicken red blood cell suspension. The mice were sacrificed 12 h later, and peritoneal macrophages were collected. The phagocytosis rate was detected by ultraviolet spectrophotometer. The results are shown in Table 2.

[0079] Table 2 Macrophage phagocytosis rate

[0080] As shown in Table 2, the fermented black ginseng products prepared in the embodiments of the present invention have significant immunomodulatory functions.

[0081] (3) Anti-fatigue function test: Swimming experiment of mice: Ninety Kunming mice were randomly divided into 9 groups (Examples 1-6, Comparative Examples 1-3), with 10 mice in each group. The dose was 200 mg / kg·d and administered by gavage for 30 consecutive days. 24 hours after the last gavage, the mice were loaded with a lead weight of 5% of their body weight on their tails and placed in a swimming tank (water temperature 25℃). The time it took for the mice to swim to exhaustion was recorded, as shown in Table 3.

[0082] Table 3 Time to Exhaustion

[0083] As shown in Table 3, the fermented black ginseng products prepared in the embodiments of the present invention have significant anti-fatigue functions.

[0084] (4) Gastrointestinal tolerance test: Experimental methods: Ninety volunteers with sensitive gastrointestinal systems were randomly divided into nine groups (Examples 1-6 and Comparative Examples 1-3), with 10 volunteers in each group. Each volunteer took 3 mL twice a day for seven consecutive days. The occurrence of symptoms such as diarrhea, bloating, and nausea during the treatment period was recorded, as shown in Table 4.

[0085] Table 4 Experimental Results

[0086] As shown in Table 4, the products of Examples 1 to 6 of the present invention have excellent intestinal tolerance, and the incidence of discomfort after taking them is extremely low in people with sensitive gastrointestinal tracts, making them suitable for a wider range of people.

[0087] (5) Taste test: One hundred volunteers without taste impairment and with the ability to identify the taste of this type of product were selected as evaluators to evaluate the taste of the products of Examples 1-6 and Comparative Examples 1-3 (with a maximum score of 10 points, the higher the score, the stronger the taste). The taste indicators included bitterness, sourness, and flavor. The results are shown in Table 5.

[0088] Among them, the flavor index in the taste score is judged by a multi-dimensional quantitative scoring method. The specific standards are as follows: (1) Aroma intensity (0-3 points): 0 points = no characteristic aroma, 1 point = weak aroma, 2 points = moderate and pure aroma, 3 points = rich and lasting aroma; (2) Taste harmony (0-4 points): 0 points = messy taste, 1 point = slightly uncoordinated taste, 2 points = harmonious taste without conflict, 3 points = distinct taste layers, 4 points = high degree of flavor integration and smooth taste; (3) Aftertaste persistence (0-3 points): 0 points = no aftertaste, 1 point = short aftertaste, 2 points = moderate aftertaste, 3 points = long and comfortable aftertaste; The scores of the above three dimensions are added together to form the final score of the flavor index (full score 10 points).

[0089] Table 5. Comparison of Taste Scores (Average Score)

[0090] Note: The flavor score is the average score based on a three-dimensional quantitative standard of "aroma intensity - taste harmony - aftertaste persistence". The higher the score, the better the flavor performance of the product.

[0091] As shown in Table 5, the overall taste scores of the products in Examples 1-6 of this invention are significantly higher than those of the respective comparative proportions, the bitterness is significantly reduced, the flavor is rich, and the problem of bitter taste in traditional black ginseng products is solved.

[0092] (6) Safety inspection: The oral liquid prepared in Example 1 was subjected to safety testing, including heavy metal (lead, mercury, cadmium) and microbial (total bacterial count, coliform bacteria, pathogenic bacteria) tests. The results are shown in Table 6.

[0093] Table 6 Safety test results

[0094] As shown in Table 6, the products of the present invention meet the national food safety standards and are highly safe.

[0095] Therefore, the present invention adopts the above-mentioned fermented black ginseng product and its preparation method, and by optimizing the fermentation process parameters, achieves efficient conversion and enrichment of the effective components of black ginseng, while reducing the irritation of the fermented black ginseng product and improving its taste, so that the fermented black ginseng product prepared is suitable for a wide range of people.

[0096] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A method for preparing a fermented black ginseng product, characterized in that, Includes the following steps: Step S1: Pre-treat the ginseng to obtain black ginseng powder; Select ginseng that is 5 years old or older, and then process it by removing impurities, washing, slicing, and steaming and drying it nine times using traditional methods to obtain ginseng slices with a moisture content of ≤7%. The slices are then pulverized and passed through a 60-90 mesh sieve to obtain black ginseng powder. Step S2: Prepare the compound fermentation broth; Step S3: Prepare the culture medium; Step S4: Inoculate and ferment to obtain black ginseng fermentation concentrate; Step S5: According to the requirements, the black ginseng fermentation concentrate and auxiliary materials are processed into shape to obtain the black ginseng fermentation product.

2. The method for preparing a fermented black ginseng product according to claim 1, characterized in that, Step S2 is as follows: Step S21: Activate and culture *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* separately. Step S22: Mix the following ingredients in a volume ratio of (2.5~3.2):(1.5~2):(1.8~2.1):(0.5~1) to obtain a compound fermentation liquid; Among them, the total viable count of the compound fermentation liquid is ≥1×10 9 CFU / mL.

3. The method for preparing a fermented black ginseng product according to claim 2, characterized in that: In step S21, the specific conditions for activating and culturing *Lactobacillus plantarum*, *Saccharomyces cerevisiae*, *Bacillus subtilis*, and *Cordyceps militaris* are as follows: Lactobacillus plantarum: MRS medium, anaerobic culture at 37℃ for 18-24 h, viable count ≥5×10⁻⁶ 9 CFU / mL; Saccharomyces cerevisiae: YPD medium, aerobic culture at 30℃ for 12-16 h, viable count ≥3×10⁻⁶ 9 CFU / mL; Bacillus subtilis: LB medium, aerobic culture at 37℃ for 16-20 h, viable count ≥4×10⁻⁶. 9 CFU / mL; Cordyceps militaris: Cultured on PDA medium at 25℃ in the dark for 48-72 hours, spore concentration ≥2×10⁻⁶. 8 CFU / mL.

4. The method for preparing a fermented black ginseng product according to claim 1, characterized in that: In step S3, the culture medium comprises the following components by weight percentage: 20-25% black ginseng powder, 3-8% fructooligosaccharides, 0.5-1.5% peptone, 0.1-0.3% KH2PO4, 0.05-0.15% MgSO4, and the remainder is deionized water.

5. The method for preparing a fermented black ginseng product according to claim 4, characterized in that, Step S3 is as follows: Step S31: Mix all components of the culture medium thoroughly and adjust the pH to 6.0~6.5; Step S32: Sterilize at 110~120℃ for 20~25 minutes, then cool to room temperature to obtain the culture medium.

6. The method for preparing a fermented black ginseng product according to claim 1, characterized in that, Step S4 is as follows: Step S41: Inoculate the culture medium with a compound fermentation broth accounting for 4-7% of the weight of the culture medium. Step S42: Perform staged gradient fermentation to obtain black ginseng fermentation liquid; Step S43: Centrifuge, extract, and concentrate the black ginseng fermentation liquid in sequence to obtain black ginseng fermentation concentrate.

7. The method for preparing a fermented black ginseng product according to claim 6, characterized in that: The phased gradient fermentation in step S42 is as follows: the temperature of the first stage of aerobic fermentation is 23~27℃, the pH is 6.0~6.5, the fermentation time is 30~40h, and the dissolved oxygen concentration is 5~8mg / L. The second stage of anaerobic fermentation is carried out at a temperature of 30-35℃, a pH of 4.8-5.0, and a fermentation time of 48-72 hours. The third stage of microaerobic fermentation is carried out at a temperature of 30-32℃, a pH of 5.5-6.0, a fermentation time of 30-35 hours, and a dissolved oxygen concentration of 1-3 mg / L.

8. The method for preparing a fermented black ginseng product according to claim 6, characterized in that: In step S43, the centrifugation speed is 6000~8000 r / min and the centrifugation time is 10~20 min; Ultrasonic extraction was performed with a power of 200-300W, an extraction temperature of 40-50℃, and an extraction time of 30-45 minutes. The concentration temperature is 50~60℃, the concentration vacuum degree is -0.08~-0.09MPa, and the solid content after concentration is 25~30%.

9. The method for preparing a fermented black ginseng product according to claim 1, characterized in that: In step S5, the fermented black ginseng product comprises the following components by weight percentage: 75-80% black ginseng fermentation concentrate and 20-25% excipients; wherein the excipients are one or more of maltitol, fructooligosaccharides, xanthan gum, and citric acid.

10. A fermented black ginseng product prepared by the method of any one of claims 1 to 9.

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