Hippophae rhamnoides synbiotics fermented beverage helpful for enhancing intestinal immunity

Abstract

The invention belongs to the field of food, and particularly relates to a preparation method of a sea-buckthorn synbiotic fermented beverage, the sea-buckthorn synbiotic fermented beverage mainly comprises the following components in percentage by weight: 0.76-2.99% of sea-buckthorn polysaccharide, 0.76-0.75% of bifidobacterium longum, 0.75-0.75% of lactobacillus acidophilus, 0.75-0.75% of sea-buckthorn fruit powder, 0.75-0.75% of skim milk powder, 0.75-0.75% of mogroside and the balance of water. The weight percentage of the bifidobacterium longum in the fermented beverage is 0.96-1.91%, the weight percentage of the lactobacillus acidophilus in the fermented beverage is 0.96-1.91%, the weight percentage of the sea buckthorn fruit powder in the fermented beverage is 0.38-2.23%, the weight percentage of the skim milk powder in the fermented beverage is 0.38-2.23%, the weight percentage of the mogroside in the fermented beverage is 0.04-0.15%, and the balance is water. The sea-buckthorn synbiotics fermented beverage disclosed by the invention is beneficial to enhancing the immunity of intestinal tracts, is suitable for a wide range of people, and is a novel functional beverage.

Description

Technical Field

[0001] The invention belongs to the field of food and relates to a method for preparing a seabuckthorn synbiotic fermented beverage which is helpful in enhancing intestinal immunity. Background Art

[0002] Synbiotics are composed of live microorganisms (probiotics) and substrates (prebiotics) that can be selectively utilized by host microorganisms. These ingredients can provide the human body with essential nutrients while ensuring food safety. In a simple and efficient way, they not only help improve digestive system problems, but also bring multiple benefits to the immune system. As the field of intestinal health continues to expand and mature, synbiotics are located at the intersection of scientific research and development and consumer demand, and its market is growing rapidly. Therefore, many companies are actively deploying synbiotics to meet the growing demand for healthy food in the market, but the application of synbiotics is not yet common. The synbiotics on the market are mostly concentrated in infant formula. There is still a huge gap in synbiotics suitable for adults, and the combination design of probiotics and prebiotics needs to consider compatibility, scientific ratios, and population adaptability. In short, the field of synbiotics still has a long way to go, and at the same time has huge development potential and room for development.

[0003] Seabuckthorn is a plant with both medicinal and edible properties. It is rich in amino acids, fatty acids, polysaccharides, vitamins, flavonoids, sterols and other bioactive substances, and has high nutritional and health value and medicinal value. Seabuckthorn polysaccharides are polysaccharides extracted from seabuckthorn, which have antioxidant, antibacterial, anticancer, blood lipid regulation, cancer and cardiovascular disease prevention effects. However, the current development and utilization of seabuckthorn in my country is mainly based on fruit juice extraction, and its main products include seabuckthorn beverages, seabuckthorn vinegar and seabuckthorn wine. After juicing, other by-products are discarded, and the beneficial components in the pomace are not effectively utilized. There are few research and development of refined seabuckthorn products, and the various components of seabuckthorn, such as flavonoids, polysaccharides and other active ingredients, have not been deeply studied and utilized. For example, the patent with application number CN202311851145.8 discloses a food for regulating the intestines and alleviating sub-health conditions and its preparation method, in which seabuckthorn is only a part of insignificant auxiliary materials, and cannot give full play to the beneficial functions of seabuckthorn. More importantly, seabuckthorn has not yet been studied in the field of synbiotics. In addition, most of the probiotics in synbiotics are anaerobic bacteria, which usually require encapsulation technology to work in the body. However, the production process of this technology is complicated and the production cost is high. In vitro fermentation can produce beneficial metabolites in vitro, ensuring the production and utilization of active substances, and effectively preventing probiotics from being destroyed in the digestive tract after oral administration and unable to reach the intestines to work.

[0004] Therefore, the present invention uses seabuckthorn polysaccharide as a prebiotic, and then uses probiotics Bifidobacterium longum and Lactobacillus acidophilus for in vitro fermentation to form a synbiotic system, and uses seabuckthorn fruit powder, skimmed milk powder and mogroside for flavoring to develop a seabuckthorn synbiotic fermented beverage, which can maximize the effect of seabuckthorn in enhancing intestinal immunity and promote the development and utilization of seabuckthorn, and has certain practical significance. Summary of the invention

[0005] In view of the deficiencies of the above existing technologies, this study provides a seabuckthorn synbiotic fermented beverage that does not add any pigments and preservatives, has a unique flavor, is rich in nutrients, and has the function of enhancing intestinal immunity.

[0006] The technical solution of the present invention is as follows: 1. A method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity, comprising the following steps: (1) Extract seabuckthorn polysaccharides by water extraction and alcohol precipitation. Fresh seabuckthorn fruit is dried at 60°C, crushed, defatted, dried and ground, and then passed through a 60-mesh sieve. The fruit is placed in distilled water with a solid-liquid ratio of 1:25. The fruit is extracted at 80°C for 1 h. The supernatant is taken by centrifugation and concentrated by rotary evaporation. The concentrate is then mixed with 4 times the volume of anhydrous ethanol and allowed to stand at 4°C for 48 h. The concentrate is then deproteinized, dialyzed, and freeze-dried to obtain seabuckthorn polysaccharides. (2) Dissolve seabuckthorn polysaccharide in distilled water to obtain seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; (3) After the seabuckthorn polysaccharide solution is cooled to room temperature, Bifidobacterium longum and Lactobacillus acidophilus are inoculated into the seabuckthorn polysaccharide solution to obtain synbiotics; (4) anaerobic fermentation of the synbiotic obtained in step (3) at 37° C. and 180 rpm on a shaking table to obtain a synbiotic fermentation liquid; (5) adding seabuckthorn fruit powder, skimmed milk powder, and mogroside to the synbiotic fermentation liquid obtained in step (4), respectively, to finally obtain a seabuckthorn synbiotic fermented beverage; Features: The weight percentage of the seabuckthorn polysaccharide in step (2) to the seabuckthorn polysaccharide solution is 0.79-3.10%; The initial concentrations of the inoculated Bifidobacterium longum and Lactobacillus acidophilus in step (3) are the same, the weight percentage of seabuckthorn polysaccharide in the synbiotic is 0.77-3.01%, the weight percentage of Bifidobacterium longum in the synbiotic is 0.96-1.93%, the weight percentage of Lactobacillus acidophilus in the synbiotic is 0.96-1.93%, and the total concentration of lactic acid bacteria in the synbiotic is 10 5 CFU / mL; The fermentation time of the synbiotic in step (4) is 6-36 h; In the synbiotic fermented beverage of step (5), the weight percentage of seabuckthorn polysaccharide in the synbiotic fermented beverage is 0.76-2.99%, the weight percentage of Bifidobacterium longum in the synbiotic fermented beverage is 0.96-1.91%, the weight percentage of Lactobacillus acidophilus in the synbiotic fermented beverage is 0.96-1.91%, the weight percentage of seabuckthorn fruit powder in the synbiotic fermented beverage is 0.38-2.23%, the weight percentage of skimmed milk powder in the synbiotic fermented beverage is 0.38-2.23%, the weight percentage of mogroside in the synbiotic fermented beverage is 0.04-0.15%, and the balance is water.

[0007] 2. A method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity, characterized in that the prepared seabuckthorn synbiotic fermented beverage has the function of assisting in enhancing intestinal immunity.

[0008] Compared with the prior art, the present invention has the following advantages: (1) The present invention uses seabuckthorn polysaccharide as a prebiotic, and Bifidobacterium longum and Lactobacillus acidophilus as probiotics. The prepared synbiotic has the function of assisting in enhancing intestinal immunity.

[0009] (2) The present invention is applicable to a wide range of people, including those with low intestinal immunity. In addition, skimmed milk powder and non-nutritive sweetener mogroside are added to the formula, making it a zero-sugar and zero-fat food, suitable for people with diabetes and dyslipidemia.

[0010] (3) No large and expensive instruments are required during production, the production cost is low, the raw material utilization rate is high, and it is suitable for large-scale production.

[0011] (4) The present invention first ferments seabuckthorn polysaccharide and bacterial powder in vitro to produce beneficial metabolites in vitro, thereby effectively reducing the damage of probiotics in the gastrointestinal tract and preventing them from exerting their biological activity.

[0012] (5) This product does not contain any pigments or preservatives. It is green, healthy, easy to carry, and can be consumed at any time. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1 The OD600 change curves of Bifidobacterium longum and Lactobacillus acidophilus in different proportions.

[0014] Figure 2 This is the mass concentration change curve of seabuckthorn polysaccharide.

[0015] Figure 3 This is the pH change curve of the synbiotic fermentation broth.

[0016] Figure 4 This is the total acid content change curve of the synbiotic fermentation broth.

[0017] Figure 5This is the curve of the viable bacterial count change in the synbiotic fermentation broth.

[0018] Figure 6 This is a picture of mouse colon.

[0019] Figure 7 Mouse colon tissue sections.

[0020] Figure 8 The mRNA expression of inflammatory factors in mouse colon. DETAILED DESCRIPTION

[0021] The present invention is further explained below with reference to examples. The embodiments are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention. Example

[0022] (1) Seabuckthorn polysaccharides were extracted by water extraction and alcohol precipitation. Fresh seabuckthorn fruit was dried and crushed at 60°C, defatted twice with petroleum ether at 60°C, and the defatted powder was dried in an oven overnight. After grinding, it was passed through a 60-mesh sieve; then placed in distilled water with a solid-liquid ratio of 1:25 and extracted at 80°C for 1 h. The extract was concentrated to one-fifth of the initial volume using a rotary evaporator, and then the concentrate was allowed to stand at 4°C for 48 h with 4 times the volume of anhydrous ethanol. The precipitate was freeze-dried and dissolved in distilled water. The protein was removed by the Savage method and then dialyzed against deionized water (1000 Da). Seabuckthorn polysaccharides were obtained after freeze-drying.

[0023] (2) Dissolve 0.4 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain seabuckthorn polysaccharide solution, and sterilize it at 70-90°C for 10-30 min.

[0024] (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 0.5 g of Bifidobacterium longum CICC 6186 and 1 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; (4) subjecting the synbiotics obtained in step (3) to constant temperature anaerobic fermentation at 37°C and 180 rpm on a shaking table for 6 h to obtain a synbiotic fermentation liquid; (5) Add 0.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside to the synbiotic fermentation liquid obtained in step (4) to season the synbiotic fermentation liquid, thereby finally obtaining a seabuckthorn synbiotic fermented beverage. Example

[0025] Step (1) and step (2) are the same as in Example 1; (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 0.6 g of Bifidobacterium longum CICC 6186 and 0.9 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; Step (4) and step (5) are the same as in Example 1. Example

[0026] Step (1) and step (2) are the same as in Example 1; (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 0.7 g of Bifidobacterium longum CICC 6186 and 0.8 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; Step (4) and step (5) are the same as in Example 1. Example

[0027] Step (1) and step (2) are the same as in Example 1; (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 0.8 g of Bifidobacterium longum CICC 6186 and 0.7 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; Step (4) and step (5) are the same as in Example 1. Example

[0028] Step (1) and step (2) are the same as in Example 1; (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 0.9 g of Bifidobacterium longum CICC 6186 and 0.6 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; Step (4) and step (5) are the same as in Example 1. Example

[0029] Step (1) and step (2) are the same as in Example 1; (3) After the seabuckthorn polysaccharide solution was cooled to room temperature, 1 g of Bifidobacterium longum CICC 6186 and 0.5 g of Lactobacillus acidophilus CICC 6074 of the same concentration were inoculated into the seabuckthorn polysaccharide solution to obtain a synbiotic. The total concentration of lactic acid bacteria in the synbiotic was 10 5 CFU / mL; Step (4) and step (5) are the same as in Example 1. Example

[0030] Step (1) is the same as in Example 1; (2) Dissolve 0.6 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3. Example

[0031] Step (1) is the same as in Example 1; (2) Dissolve 0.8 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3. Example

[0032] (Step (1) is the same as in Example 1; (2) Dissolve 1 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3.

[0033] Example 10 Step (1) is the same as in Example 1; (2) Dissolve 1.2 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3.

[0034] Embodiment 11 Step (1) is the same as in Example 1; (2) Dissolve 1.4 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3.

[0035] Example 12 Step (1) is the same as in Example 1; (2) Dissolve 1.6 g of seabuckthorn polysaccharide in 50 g of distilled water to obtain a seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; Steps (3) to (5) are the same as those in Example 3.

[0036] Embodiment 13 Steps (1) to (3) are the same as those in Example 9; (4) The temperature was 37°C and the shaking speed was 180 rpm for 12 h to obtain the synbiotic fermentation liquid; Step (5) is the same as in Example 1.

[0037] Embodiment 14 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm on a shaking table for 18 h to obtain the synbiotic fermentation liquid; Step (5) is the same as in Example 1.

[0038] Embodiment 15 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 24 h to obtain the synbiotic fermentation liquid; Step (5) is the same as in Example 1.

[0039] Example 16 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 30 h to obtain the synbiotic fermentation liquid; Step (5) is the same as in Example 1.

[0040] Embodiment 17 Steps (1) to (3) are the same as those in Example 9; (4) The temperature was 37°C and the shaking speed was 180 rpm for 36 h to obtain the synbiotic fermentation liquid; Step (5) is the same as in Example 1.

[0041] Embodiment 18 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.4 g seabuckthorn fruit powder, 0.2 g skimmed milk powder, and 0.02 g mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0042] Embodiment 19 Steps (1) to (4) are the same as in Example 18; (5) 0.6 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0043] Embodiment 20 Steps (1) to (4) are the same as in Example 18; (5) 0.8 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0044] Embodiment 21 Steps (1) to (4) are the same as in Example 18; (5) 1 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0045] Embodiment 22 Steps (1) to (4) are the same as in Example 18; (5) 1.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0046] Embodiment 23 Steps (1) to (4) are the same as in Example 18; (5) 0.2 g of seabuckthorn fruit powder, 0.4 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0047] Embodiment 24 Steps (1) to (4) are the same as in Example 18; (5) 0.2 g of seabuckthorn fruit powder, 0.6 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0048] Embodiment 25 Steps (1) to (4) are the same as in Example 18; (5) 0.2 g of seabuckthorn fruit powder, 0.8 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0049] Embodiment 26 Steps (1) to (4) are the same as in Example 18; (5) 0.2 g of seabuckthorn fruit powder, 1 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0050] Embodiment 27 Steps (1) to (4) are the same as in Example 18; 0.2 g of seabuckthorn fruit powder, 1.2 g of skimmed milk powder and 0.02 g of mogroside were added respectively to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0051] Embodiment 28 Steps (1) to (4) are the same as in Example 18; (5) 0.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0052] Embodiment 29 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.04 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0053] Embodiment 30 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.06 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0054] Embodiment 31 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.2 g of seabuckthorn fruit powder, 0.2 g of skimmed milk powder, and 0.08 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0055] Embodiment 32 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.4 g of seabuckthorn fruit powder, 0.4 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0056] Embodiment 33 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.6 g of seabuckthorn fruit powder, 0.6 g of skimmed milk powder, and 0.04 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0057] Embodiment 34 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 18 h to obtain the synbiotics. (5) 0.8 g of seabuckthorn fruit powder, 0.8 g of skimmed milk powder, and 0.06 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0058] Embodiment 35 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 24 h to obtain the synbiotics. (5) 0.6 g of seabuckthorn fruit powder, 0.8 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0059] Embodiment 36 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 24 h to obtain the synbiotics. (5) 0.8 g of seabuckthorn fruit powder, 0.4 g of skimmed milk powder, and 0.04 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0060] Embodiment 37 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 24 h to obtain the synbiotics. (5) 0.4 g seabuckthorn fruit powder, 0.6 g skimmed milk powder, and 0.06 g mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0061] Embodiment 38 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 30 h to obtain the synbiotics. (5) 0.8 g of seabuckthorn fruit powder, 0.6 g of skimmed milk powder, and 0.02 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0062] Embodiment 39 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 30 h to obtain the synbiotics. (5) 0.4 g seabuckthorn fruit powder, 0.8 g skimmed milk powder, and 0.04 g mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0063] Embodiment 40 Steps (1) to (3) are the same as those in Example 9; (4) Anaerobic fermentation was carried out at 37°C and 180 rpm in a shaking incubator for 30 h to obtain the synbiotics. (5) 0.6 g of seabuckthorn fruit powder, 0.4 g of skimmed milk powder, and 0.06 g of mogroside were added to flavor the synbiotic fermentation liquid, and finally a seabuckthorn synbiotic fermented beverage was obtained.

[0064] Experimental Example 1 Preparation of Synbiotic Fermentation Broth (1) Bacterial strain expansion culture Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074 were taken out from the -80℃ refrigerator, and the bacterial liquid was aspirated and inoculated into PYG liquid medium and MRS liquid medium respectively for strain recovery. The volume ratio of bacterial liquid to medium was 1:10. The culture was carried out in an anaerobic incubator at 37℃ for 24 hours. After successful recovery, the culture was then subcultured at a 2-4% (v / v) inoculation amount. After 24 hours of amplification culture, the culture was centrifuged at 3000 rpm and 4℃ for 10 minutes, the supernatant was discarded, and the culture was washed with PBS buffer. This was repeated 2-3 times to prepare a bacterial suspension. After measurement and dilution, the bacterial liquid concentration was ensured to reach 10 9 CFU / mL, and then the bacterial solution was freeze-dried to prepare bacterial powder for later use.

[0065] (2) Determination of the optimal mixing ratio of Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074 Examples 1 to 6 are explorations of the optimal mixing ratio of Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074. A series of gradients are set in the range of 1:2-2:1 according to the bacterial population ratio of Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074, and the initial concentration of the two bacterial solutions is ensured to be 10 9 CFU / mL, seabuckthorn polysaccharide was used as the sole carbon source, the temperature was set at 37°C, the measurement time was 42 h, automatic shaking was performed every 1 h, and the OD600 value was measured.

[0066] Determine the optimal mixing ratio of the two bacteria, such as Figure 1 As shown in the figure, it can be seen that when the number ratio of Bifidobacterium longum CICC 6186 to Lactobacillus acidophilus CICC 6074 is 2:3, 7:8 and 8:7, the OD600 value is higher. When the number ratio of Bifidobacterium longum CICC 6186 to Lactobacillus acidophilus CICC 6074 is 7:8, the OD600 value is the highest, indicating that the optimal mixing ratio is 7:8, that is, Example 3.

[0067] (3) Exploration of the optimal amount of seabuckthorn polysaccharide added Examples 3, 7 to 12 are explorations of the optimal amount of seabuckthorn polysaccharide added. Seabuckthorn polysaccharide and bacterial solution with different mass concentration gradients (0.8-3.2% w / v) were added to the MRS liquid culture medium without carbon source. Two time groups were set. After anaerobic constant temperature culture for 24 hours, the 24-hour group was taken out to measure the pH value; after another constant temperature culture for 24 hours, the pH value was measured and the pH value change curve was drawn.

[0068] The pH value change curve is as follows Figure 2 As shown in the figure, it can be seen that the optimum range of the mass concentration of seabuckthorn polysaccharide is 1.2-2.4%, that is, the optimum range is Example 7 to Example 10, and the best mass concentration of seabuckthorn polysaccharide is 2.0%, that is, Example 9.

[0069] (4) Exploration of the fermentation time of seabuckthorn polysaccharides and bacteria Example 9 and Example 13 to Example 17 are explorations of the fermentation time of seabuckthorn polysaccharides and bacteria. Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074 were added to a seabuckthorn polysaccharide solution with a seabuckthorn polysaccharide mass concentration of 2.0% (w / v) at an optimal mixing ratio of 7:8, and the fermentation was carried out at a constant temperature of 37°C and 180 rpm on a shaking table. The pH value and total acid content of the fermentation liquid were determined every 6 hours according to the method specified in GB 12456-2021 "National Food Safety Standard Determination of Total Acid in Foods" until 36 hours.

[0070] The pH value change curve of synbiotic fermentation broth is as follows Figure 3 As shown in the figure, it can be seen that the pH value of the synbiotic fermentation liquid is relatively stable in the first 24 hours of fermentation time, and then the pH value decreases rapidly; in addition, the total acid content change curve of the synbiotic fermentation liquid is as shown in Figure 4 As shown, the results are similar to the pH value. Considering the comprehensive fermentation time, it is more suitable when it is 18-30 h, that is, the optimal range is Example 14 to Example 16.

[0071] Test Example Determination of viable bacterial count in synbiotic fermentation broth Bifidobacterium longum CICC 6186 and Lactobacillus acidophilus CICC 6074 were added to a sea buckthorn polysaccharide solution with a mass concentration of 2.0% (w / v) at an optimal mixing ratio of 7:8, and the mixture was anaerobic fermented at 37°C and 180 rpm in a shaking table. The number of viable lactic acid bacteria in the solution was detected at 0 h, 6 h, 12 h, 18 h, 24 h, 30 h, and 36 h of fermentation, respectively, according to GB 4789.35-2016 "Food Microbiology Test - Test of Lactic Acid Bacteria".

[0072] The number of viable bacteria in synbiotic fermentation broth is Figure 5 As shown in the figure, it can be seen that after 18 hours of fermentation, the number of viable bacteria in the fermentation liquid reached 10 7 CFU / mL, and as the fermentation time increases, the number of live bacteria in the synbiotic fermentation broth increases.

[0073] Experimental Example 3 Sensory Evaluation of Seabuckthorn Synbiotic Fermented Beverage The sensory evaluation of the seabuckthorn synbiotic fermented beverage was carried out with a full score of 100. The evaluation panel consisted of 10 people with certain sensory evaluation experience. Each person gave their own score and the average value was taken. The evaluation was mainly based on the color, flavor, smell, taste and texture of the synbiotic fermented beverage, with each accounting for 20% of the total score for color, 20% for flavor, 20% for smell, 20% for taste and 20% for texture. The sensory evaluation standards and results are shown in Tables 1 and 2: Table 1 Sensory evaluation criteria Table 2 Sensory evaluation results of synbiotic fermented beverages project Color Flavor odor Taste Organization Status Total score Embodiment 18 The color is relatively uniform, turbid, and brown (12 points) Too sour or too sweet, with a unique flavor of sea buckthorn (13 points) Light milk flavor (13 points) The taste is bland, with a little astringency (12 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 65 points Embodiment 19 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Light milk flavor (13 points) The taste is bland, with a little astringency (12 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 67 points Embodiment 20 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (13 points) Light milk flavor (14 points) The taste is bland, with a little astringency (13 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 68 points Embodiment 21 The color is relatively uniform, turbid, and brown (11 points) Too sour or too sweet, with a unique flavor of sea buckthorn (11 points) Light milk flavor (13 points) The taste is bland, with a little astringency (11 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 61 points Embodiment 22 The color is relatively uniform, turbid, and brown (11 points) Too sour or too sweet, with a unique flavor of sea buckthorn (11 points) Light milk flavor (13 points) The taste is bland, with a little astringency (12 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 62 points Embodiment 23 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (13 points) Light milk flavor (12 points) The taste is relatively round, without astringency (15 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 68 points Embodiment 24 The color is relatively uniform, turbid, and brown (14 points) Too sour or too sweet, with a unique flavor of sea buckthorn (13 points) Light milk flavor (12 points) The taste is bland, with a little astringency (14 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 68 points Embodiment 25 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (13 points) Light milk flavor (13 points) The taste is bland, with a little astringency (14 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 68 points Embodiment 26 The color is relatively uniform, turbid, and brown (11 points) Too sour or too sweet, with a unique flavor of sea buckthorn (12 points) Light milk flavor (12 points) The taste is bland, with a little astringency (13 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 63 points Embodiment 27 Very cloudy, too dark or too light (10 points) Too sour or too sweet, with a unique flavor of sea buckthorn (11 points) Light milk flavor (13 points) The taste is bland, with a little astringency (11 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 60 points Embodiment 28 The color is relatively uniform, turbid, and brown (14 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Light milk flavor (13 points) The taste is bland, with a little astringency (12 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 68 points Embodiment 29 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Moderate milk flavor (15 points) The taste is bland, with a little astringency (12 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 69 points Embodiment 30 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Light milk flavor (13 points) The taste is bland, with a little astringency (14 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 69 points Embodiment 31 The color is relatively uniform, turbid, and brown (12 points) Too sour or too sweet, lacking the unique flavor of sea buckthorn (10 points) Light milk flavor (11 points) Tastes bitter, too sweet or too sour (10 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 58 points Embodiment 32 The color is relatively uniform, shiny, and milky coffee color (16 points) Sour or sweet, with a unique flavor of sea buckthorn (16 points) Moderate milk flavor (16 points) The taste is relatively round, without astringency (15 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 78 points Embodiment 33 The color is relatively uniform, shiny, and milky coffee color (15 points) Sour or sweet, with a unique flavor of sea buckthorn (16 points) Moderate milk flavor (16 points) The taste is relatively round, without astringency (15 points) The texture is relatively fine, clear, without precipitation and impurities (16 points) 78 points Embodiment 34 The color is relatively uniform, shiny, and milky coffee color (15 points) Sour or sweet, with a unique flavor of sea buckthorn (15 points) Moderate milk flavor (15 points) The taste is relatively round, without astringency (16 points) The texture is relatively fine, clear, without precipitation and impurities (16 points) 77 points Embodiment 35 The color is relatively uniform, turbid, and brown (17 points) Sour or sweet, with a unique flavor of sea buckthorn (15 points) Moderate milk flavor (17 points) The taste is round and not astringent (17 points) Fine texture, good solubility, no precipitation, no impurities (17 points) 83 points Embodiment 36 The color is uniform and shiny, and is milky coffee color (19 points) The taste is sweet and sour, with a unique flavor of sea buckthorn (19 points) Moderate milk flavor (17 points) The taste is relatively round, without astringency (19 points) The texture is relatively fine, clear, without precipitation and impurities (20 points) 94 points Embodiment 37 The color is relatively uniform, turbid, and brown (14 points) Sour or sweet, with a unique flavor of sea buckthorn (17 points) Moderate milk flavor (16 points) The taste is relatively round, without astringency (17 points) The texture is relatively fine, clear, without precipitation and impurities (15 points) 79 points Embodiment 38 The color is relatively uniform, shiny, and milky coffee color (15 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Light milk flavor (14 points) The taste is relatively round, without astringency (15 points) The tissue is slightly turbid, with a small amount of precipitation and a small amount of impurities (14 points) 72 points Embodiment 39 The color is relatively uniform, turbid, and brown (14 points) Sour or sweet, with a unique flavor of sea buckthorn (15 points) Moderate milk flavor (15 points) The taste is bland, with a little astringency (14 points) The tissue is slightly turbid, with a small amount of precipitation and a small amount of impurities (13 points) 71 points Embodiment 40 The color is relatively uniform, turbid, and brown (13 points) Too sour or too sweet, with a unique flavor of sea buckthorn (14 points) Moderate milk flavor (15 points) The taste is bland, with a little astringency (14 points) The tissue is slightly turbid, with a small amount of precipitation and a small amount of impurities (13 points) 69 points The seabuckthorn synbiotic fermented beverage is a milky coffee-colored liquid. The color, flavor, smell, taste and texture of the synbiotic fermented beverage were evaluated according to the sensory evaluation standards in Table 1. The results are shown in Table 2. Examples 18 to 22 are explorations of the amount of seabuckthorn fruit powder added, among which the amount of seabuckthorn fruit powder added in Examples 18 to 20 is more appropriate; Examples 23 to 27 are explorations of the amount of skim milk powder added, among which Examples 23 to 25 are more appropriate ranges; Examples 28 to 31 are explorations of the amount of mogroside added, among which Examples 28 to 30 are more appropriate.

[0074] In combination with the preferred ranges of the above embodiments, a four-factor three-level orthogonal test was carried out on the addition amount of sea buckthorn fruit powder, skimmed milk powder, mogroside and fermentation time, specifically Examples 32 to 40. From the sensory score, it can be seen that the raw material ratio of Example 36 is the optimal ratio. The synbiotic fermented beverage obtained by this ratio has a uniform and shiny color, a milky coffee color, appropriate sweet and sour taste, moderate milk flavor, a mellow taste, no astringency, and has a unique flavor of sea buckthorn, fine texture, good solubility, no precipitation, and no impurities.

[0075] Experimental Example 4 Analysis of Seabuckthorn Synbiotic Fermented Beverage to Enhance Immune Function Material A seabuckthorn synbiotic fermented beverage prepared according to the recipe of Example 36.

[0076] Animal husbandry The purchased 8-week-old male mice (C57BL / 6J) were adaptively fed for 1 week and then randomly divided into groups (12 in each group). The specific groups were as follows: ① Control group: gavage with 0.2 mL PBS; ② DSS group: gavage with 0.2 mL PBS + 3% (w / v) DSS solution; ③ DSS + synbiotic group: gavage with 0.2 mL synbiotic fermented beverage + 3% (w / v) DSS solution. The mice in each group were raised under standard conditions (temperature 22±2℃, humidity 50±15%, 12 h / 12 ​​h light-dark cycle) and fed with standard feed. They were intervened with PBS and synbiotic fermented beverage for one month, and then the colitis model was established with 3% (w / v) DSS solution for one week.

[0077] Disease Activity Index Assessment In order to evaluate the severity of DSS-induced colitis, the weight, stool viscosity and stool bleeding of the mice were evaluated from the first day of modeling to the end of the experiment. The scores obtained for each scoring criterion were judged separately, and finally the average of the three scores was taken as the final disease activity index, which was used to judge the severity of colitis in the experimental mice.

[0078] Hematoxylin and eosin (H&E) staining The mouse colon tissue was carefully collected, fixed in 4% paraformaldehyde and embedded in paraffin. The tissue was cut into 4 μm thick sections and stained with hematoxylin and eosin, and observed and photographed under an optical microscope.

[0079] Alcian blue staining The colon was fixed on a slide (5 μm), dewaxed with xylene and rehydrated in ethanol, then stained with Alcian blue solution for 30 min. The slides were washed in running tap water for 2 min and rinsed with distilled water and observed under a microscope.

[0080] Real-time quantitative polymerase chain reaction (RT-qPCR) The colon tissue was ground and the RNA in the intestinal tissue was extracted using Trizol reagent. The concentration of the extracted RNA was determined by measuring the OD value using a nucleic acid quantifier, and its purity was determined. The RNA was reverse transcribed into cDNA using a reverse transcription kit. A real-time PCR reaction system was established for PCR amplification. GAPDH was standardized to the CT value, and the relative expression of each gene was calculated using the ΔΔCT method.

[0081] Experimental Results (1) Effect of seabuckthorn synbiotic fermented beverage on body weight of mice During the DSS modeling period, the body weight of the mice was recorded every day and the weight gain was calculated. The results are shown in Table 3.

[0082] Table 3 Average body weight of mice in each group Note: Compared with the control group, **p<0.01 indicates an extremely significant difference; compared with the DSS group, #p<0.05 indicates a significant difference, and ##p<0.01 indicates an extremely significant difference.

[0083] It can be seen from Table 3 that compared with the control group, the DSS group lost weight significantly 3 days after modeling, while the DSS+synbiotic group lost less weight, indicating that synbiotic fermented beverage has a certain improvement effect on the weight loss of mice caused by DSS.

[0084] (2) Effect of seabuckthorn synbiotic fermented beverage on colon length in mice After the mice were sacrificed, the colons of the mice were photographed and the length of the colon was recorded. Figure 6 As shown in Table 4.

[0085] Table 4 Colon length of mice in each group Note: Compared with the control group, **p<0.01 indicates an extremely significant difference; compared with the DSS group, ##p<0.01 indicates an extremely significant difference.

[0086] Depend on Figure 6As shown in Table 4, the colon length of mice in the DSS group was significantly shorter than that in the control group, and the colon length of mice in the synbiotic fermented beverage intervention group was significantly longer than that in the DSS group, indicating that the synbiotic fermented beverage can effectively prevent the shortening of the colon length of mice with colitis.

[0087] (3) Effect of seabuckthorn synbiotic fermented beverage on disease activity index in mice The disease activity index of mice was evaluated based on the body weight, stool consistency and intestinal bleeding of colitis mice. The results are shown in Table 5.

[0088] Table 5 Disease activity index of mice in each group Note: Compared with the control group, **p<0.01 indicates an extremely significant difference; compared with the DSS group, ##p<0.01 indicates an extremely significant difference.

[0089] The disease activity index is used to evaluate the severity of DSS-induced colitis. As shown in Table 5, the disease activity index of the DSS group was high, and the disease activity index of the synbiotic fermented beverage intervention group was significantly lower than that of the DSS group from the fourth day to the sixth day, indicating that the severity of colitis in the DSS group gradually increased with time, and was the most severe on the sixth day. After the intervention of the synbiotic fermented beverage, the severity of colitis was significantly alleviated, and the disease activity index on the fifth day was the largest difference from the DSS group.

[0090] (4) Effects of seabuckthorn synbiotic fermented beverage on the colon tissue morphology of mice DSS can cause serious damage to the intestinal barrier of mice with colitis. H&E staining and Alcian blue staining were used to investigate the effect of seabuckthorn synbiotic fermented beverage on the colon tissue morphology of mice. Figure 7 As shown, DSS treatment led to a significant reduction in mucus in the colon tissue of mice, direct exposure of the muscular stratum, and a significant reduction in intestinal goblet cells. The mice in the synbiotic fermented beverage intervention group showed improvement compared with the DSS group.

[0091] (5) Effects of seabuckthorn synbiotic fermented beverage on colon inflammatory factors in mice mRNA expression of inflammatory factors in the colon Figure 8 As shown, compared with the control group, the mRNA expression of inflammatory factors IL-1β, IL-6, and TNF-α in the DSS group was significantly increased, while the mRNA expression of inflammatory factors was significantly restored after intervention with synbiotic fermented beverage, indicating that seabuckthorn synbiotic beverage can inhibit the expression of inflammatory factors in the intestine and enhance intestinal immunity.

[0092] Based on the equivalent dose ratio between humans and animals based on the body surface area, the oral dose of 0.2 mL for mice is 39 mL / d relative to that for humans (taking 60 kg body weight as an example). The recommended daily dose of Example 36 for humans is once a day, 39 mL each time.

Claims

1. A method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity, comprising the following steps: (1) Extract seabuckthorn polysaccharides by water extraction and alcohol precipitation. Fresh seabuckthorn fruit is dried at 60°C, crushed, defatted, dried and ground, and then passed through a 60-mesh sieve. The fruit is placed in distilled water with a solid-liquid ratio of 1:

25. The fruit is extracted at 80°C for 1 h. The supernatant is taken by centrifugation and concentrated by rotary evaporation. The concentrate is then mixed with 4 times the volume of anhydrous ethanol and allowed to stand at 4°C for 48 h. The concentrate is then deproteinized, dialyzed, and freeze-dried to obtain seabuckthorn polysaccharides. (2) Dissolve seabuckthorn polysaccharide in distilled water to obtain seabuckthorn polysaccharide solution, and sterilize the seabuckthorn polysaccharide solution at 70-90°C for 10-30 min; (3) After the seabuckthorn polysaccharide solution is cooled to room temperature, Bifidobacterium longum and Lactobacillus acidophilus are inoculated into the seabuckthorn polysaccharide solution to obtain synbiotics; (4) anaerobic fermentation of the synbiotic obtained in step (3) at 37° C. and 180 rpm on a shaking table to obtain a synbiotic fermentation liquid; (5) adding seabuckthorn fruit powder, skimmed milk powder, and mogroside to the synbiotic fermentation liquid obtained in step (4), respectively, to finally obtain a seabuckthorn synbiotic fermented beverage; Features: The weight percentage of the seabuckthorn polysaccharide in step (2) to the seabuckthorn polysaccharide solution is 0.79-3.10%; The initial concentrations of the inoculated Bifidobacterium longum and Lactobacillus acidophilus in step (3) are the same, the weight percentage of seabuckthorn polysaccharide in the synbiotic is 0.77-3.01%, the weight percentage of Bifidobacterium longum in the synbiotic is 0.96-1.93%, the weight percentage of Lactobacillus acidophilus in the synbiotic is 0.96-1.93%, and the total concentration of lactic acid bacteria in the synbiotic is 10 5 CFU / mL; The fermentation time of the synbiotic in step (4) is 6-36 h; In the synbiotic fermented beverage of step (5), the weight percentage of seabuckthorn polysaccharide in the synbiotic fermented beverage is 0.76-2.99%, the weight percentage of Bifidobacterium longum in the synbiotic fermented beverage is 0.96-1.91%, the weight percentage of Lactobacillus acidophilus in the synbiotic fermented beverage is 0.96-1.91%, the weight percentage of seabuckthorn fruit powder in the synbiotic fermented beverage is 0.38-2.23%, the weight percentage of skimmed milk powder in the synbiotic fermented beverage is 0.38-2.23%, the weight percentage of mogroside in the synbiotic fermented beverage is 0.04-0.15%, and the balance is water.

2. The method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity according to claim 1, characterized in that: The weight percentage of the seabuckthorn polysaccharide in step (2) to the seabuckthorn polysaccharide solution is 1.19-2.34%; The initial concentrations of the inoculated Bifidobacterium longum and Lactobacillus acidophilus in step (3) are the same, the weight percentage of seabuckthorn polysaccharide in the synbiotic is 1.15-2.28%, the weight percentage of Bifidobacterium longum in the synbiotic is 1.16-1.54%, the weight percentage of Lactobacillus acidophilus in the synbiotic is 1.35-1.73%, and the total concentration of lactic acid bacteria in the synbiotic is 10 5 CFU / mL; The fermentation time of the synbiotic in step (4) is 18-30 h; In the synbiotic fermented beverage of step (5), the weight percentage of seabuckthorn polysaccharide in the synbiotic fermented beverage is 1.14-2.26%, the weight percentage of Bifidobacterium longum in the synbiotic fermented beverage is 1.15-1.53%, the weight percentage of Lactobacillus acidophilus in the synbiotic fermented beverage is 1.34-1.72%, the weight percentage of seabuckthorn fruit powder in the synbiotic fermented beverage is 0.74-1.86%, the weight percentage of skimmed milk powder in the synbiotic fermented beverage is 0.74-1.86%, the weight percentage of mogroside in the synbiotic fermented beverage is 0.04-0.11%, and the balance is water.

3. The method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity according to claim 1, characterized in that: The weight percentage of seabuckthorn polysaccharide in the seabuckthorn polysaccharide solution in step (2) is 1.96%; The initial concentrations of the inoculated Bifidobacterium longum and Lactobacillus acidophilus in step (3) are the same, the weight percentage of seabuckthorn polysaccharide in the synbiotic is 1.90%, the weight percentage of Bifidobacterium longum in the synbiotic is 1.33%, the weight percentage of Lactobacillus acidophilus in the synbiotic is 1.52%, and the total concentration of lactic acid bacteria in the synbiotic is 10 5 CFU / mL; The fermentation time of the synbiotic in step (4) is 24 h; In step (5), the weight percentage of seabuckthorn polysaccharide in the synbiotic fermented beverage is 1.86%, the weight percentage of Bifidobacterium longum in the synbiotic fermented beverage is 1.30%, the weight percentage of Lactobacillus acidophilus in the synbiotic fermented beverage is 1.49%, the weight percentage of seabuckthorn fruit powder in the synbiotic fermented beverage is 1.49%, the weight percentage of skimmed milk powder in the synbiotic fermented beverage is 0.74%, the weight percentage of mogroside in the synbiotic fermented beverage is 0.07%, and the balance is water.

4. The method for preparing a seabuckthorn synbiotic fermented beverage that helps enhance intestinal immunity according to any one of claims 1 to 3, characterized in that: The prepared seabuckthorn synbiotic fermented beverage has the function of assisting in enhancing intestinal immunity.

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