Health-care composition containing microbial beneficial bacteria and preparation method of health-care composition

Abstract

The invention relates to the technical field of health-care food, and discloses a health-care composition containing microbial beneficial bacteria and a preparation method of the health-care composition. Comprising the following components in parts by weight: 30-40 parts of gelatinized starch, 8-15 parts of plant enzyme powder, 2-10 parts of microcapsule compound beneficial bacteria, 3-5 parts of selenium-enriched yeast powder, 0.5-2 parts of an American ginseng extract, 12-20 parts of fructo-oligosaccharide, 1-2.5 parts of curcumin, 7-9 parts of ovalbumin and 5-8 parts of sodium carboxymethyl cellulose. Microcapsule composite beneficial bacteria are prepared by compounding bifidobacterium longum, lactobacillus acidophilus, lactobacillus rhamnosus and lactococcus lactis, bridging with carboxymethylated beta-glucan and a Fe < 3 + >-polyphenol network structure and embedding with a protein-polysaccharide system, and can stably exist in an environment with strong acid and high-concentration bile salt to gastrointestinal tracts; through the synergistic effect of different nutrient substances, the intestinal microbe balance can be improved, and the immunity can be remarkably improved.

Description

Technical Field

[0001] The invention relates to the technical field of health food, in particular to a health care composition containing beneficial microorganisms and a preparation method thereof. Background Art

[0002] Beneficial microorganisms, also known as microecological regulators, are non-pathogenic microorganisms that are active and can survive independently. As microecological regulators, they have the functions of balancing intestinal flora, relieving intestinal inflammation and improving human intestinal health. When the human body lacks beneficial bacteria, it will be in a sub-healthy state and is prone to a series of symptoms such as diarrhea, allergies, poor appetite, fatigue, and low immunity. Therefore, the proper use of beneficial bacteria can help to balance the flora in the body. According to the general concept of the application effect of beneficial bacteria, only live bacteria can produce probiotic functions. Therefore, beneficial bacteria must be alive when ingested and remain active in the gastrointestinal tract. However, beneficial bacteria in the prior art are generally added to beverages and foods, and their product types are mainly powders, but powders need to be consumed immediately after being rinsed with water, have poor adhesion in the intestine, short duration, and are difficult to maintain the balance of intestinal flora for a long time.

[0003] At present, most of the beneficial bacteria health products on the market add inactivated bacteria or beneficial bacteria with poor stability and small number, which leads to complete inactivation of beneficial bacteria within the effective period, and the corresponding effect cannot be achieved by taking health products; at the same time, since beneficial bacteria are more sensitive to the environment of strong acid and high concentration of bile salt in the gastrointestinal tract, it is difficult to survive and colonize after digestion in the gastrointestinal tract, and the adhesion to the intestinal wall is poor, and the colonization efficiency is low, which further affects the efficacy of beneficial bacteria. In order to improve the shortcomings of beneficial bacteria such as acid resistance and poor long-term storage stability, patent number CN105310080B discloses beneficial bacteria microcapsules and their preparation method and application, the beneficial bacteria microcapsules include core material and wall material, the core material is beneficial bacteria, and the outer layer of the wall material is coated with chitosan; the wall material is prepared from an aqueous solution containing natural polymer materials and freeze-drying protective agents, but its beneficial bacteria are added in a single type, and have poor adhesion in the intestine, short duration, and limited effect.

[0004] There are currently a variety of methods to help beneficial bacteria survive and multiply while taking health products, and to improve the intestinal colonization efficiency of beneficial bacteria, such as extrusion, freeze drying and spray drying, etc., although these methods have improved the activity of beneficial bacteria in the intestine to a certain extent, there are problems such as complex operation and low yield, and mechanical damage may be caused to the beneficial bacteria. Therefore, the main purpose of the present invention is to wrap the composite beneficial bacteria layer by layer by a simple and effective method to enhance the resistance of the beneficial bacteria to the stress of the gastrointestinal environment, prolong the retention time of the beneficial bacteria in the intestine, and compound with other nutrients to form a health product, which can supplement the nutrition required by the human body and enhance the resistance and immunity of the body. Summary of the invention

[0005] The purpose of the present invention is to solve the problems existing in the above-mentioned prior art and to provide a health care composition containing beneficial microorganisms and a preparation method thereof, so that the beneficial microorganisms have the ability to resist acid and choline, and there is no safety concern about consumption, the adhesion of the beneficial bacteria in the intestine is improved, and the composition has a long-term sustained release, which can effectively maintain the balance of intestinal flora.

[0006] The present invention is achieved through the following technical solutions:

[0007] A health care composition containing beneficial microorganisms comprises the following components in parts by weight: 30-40 parts of gelatinized starch, 8-15 parts of plant enzyme powder, 2-10 parts of microcapsule composite beneficial bacteria, 3-5 parts of selenium-enriched yeast powder, 0.5-2 parts of American ginseng extract, 12-20 parts of oligofructose, 1-2.5 parts of curcumin, 7-9 parts of ovalbumin, and 5-8 parts of sodium carboxymethyl cellulose.

[0008] The preparation method of the health care composition containing beneficial microorganisms comprises the following steps: adding egg white protein and sodium carboxymethyl cellulose into deionized water, stirring and dissolving, adjusting the pH to 2.5-3.6 with a 25% hydrochloric acid aqueous solution, adding curcumin and microcapsule composite beneficial bacteria, stirring evenly at 20-30 DEG C and 300-500r / min, then adding gelatinized starch, plant enzyme powder, selenium-enriched yeast powder, American ginseng extract and oligofructose, mixing and heating, stirring evenly, freeze-drying, and pressing into tablets to obtain the health care composition containing beneficial microorganisms.

[0009] The preparation method of American ginseng extract is as follows: slice the American ginseng raw medicinal material, add 10 times the amount of edible ethanol, soak at 25°C for 1 hour, heat under reflux for 2 hours, and obtain the extract after concentration and spray drying.

[0010] Preferably, the preparation method of gelatinized starch is: mix sweet potato starch and deionized water in a mass ratio of 1:2, adjust the pH to 6-7 with 5% sodium carbonate aqueous solution, and heat in a water bath at 90-100° C. until completely formed into a paste.

[0011] Preferably, the preparation method of plant enzyme powder is: steam bitter gourd and apple in a mass ratio of 2:3 at 100°C for 10 minutes, cool to 30°C, add to a mixture of brown rice enzyme and soybean enzyme in a mass ratio of 1:1 under sterile conditions, and ferment at 32°C for 30 hours.

[0012] Preferably, the mixing heating temperature is 40-50° C., the stirring speed is 800-1000 r / min, and the stirring time is 2-5 h.

[0013] Preferably, the preparation method of microcapsule composite beneficial bacteria is:

[0014] Step (1), inoculating the preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis into corresponding liquid culture medium, culturing continuously for 16-32 hours at a temperature of 30-50° C. and a rotation speed of 150-250 r / min, and mixing evenly to obtain composite microbial beneficial bacteria;

[0015] Step (2), placing the composite microbial beneficial bacteria in a liquid culture medium, culturing at a constant temperature of 37° C. overnight, resuspending in a PBS buffer to obtain a composite microbial beneficial bacteria suspension, adding a polyphenol compound and FeCl3·6H2O, reacting at room temperature for 8-16 hours, high-speed centrifuging at 5000-6000 r / min for 3-5 minutes, adding carboxymethylated β-glucan for coating, washing with a PBS buffer, and freeze-drying to obtain microcapsule composite beneficial bacteria.

[0016] Preferably, in step (1), the mass ratio of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis is 3:3:2:2.

[0017] Preferably, the polyphenol compound in step (2) is any one of tannic acid, anthocyanin or lignin polyphenol.

[0018] Preferably, the viable bacterial count of the composite microbial beneficial bacteria suspension in step (2) is 4×10 10 CFU / g; the volume ratio of the composite microbial beneficial bacteria suspension, the polyphenol compound solution, FeCl3·6H2O, and the carboxymethylated β-glucan is 100:20:20:10.

[0019] Preferably, in step (2), the mass concentrations of the polyphenol compound, FeCl3·6H2O, and carboxymethylated β-glucan are 50 mg / mL, 15 mg / mL, and 0.2 mg / mL, respectively.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] (1) Carboxymethylated β-glucan has a macroporous honeycomb structure, which can more effectively capture beneficial microorganisms. At the same time, phenolic substances have more hydroxyl groups and higher adhesion ability, which can adsorb carboxymethylated β-glucan to Fe through hydrogen bonding. 3+-The surface of the polyphenol network structure is used to achieve the purpose of encapsulating the beneficial bacteria of the complex; ovalbumin, as the most abundant phospholipid-containing glycoprotein in egg white, has good nutritional value and functional properties, and can improve the taste of food; sodium carboxymethyl cellulose is an anionic polysaccharide with good gelation effect, and can produce electrostatic adsorption with the positively charged ovalbumin, effectively encapsulating the microencapsulated composite beneficial bacteria and curcumin, forming a protein-polysaccharide coated nanosystem, thereby effectively encapsulating the composite beneficial bacteria and curcumin. Therefore, after multi-layer encapsulation, the composite beneficial bacteria can stably exist in the environment of strong acid and high concentration of bile salts in the gastrointestinal tract, and better play its functional role.

[0022] (2) The protein-polysaccharide coated nanosystem in the outer layer has high hydrophilicity and adhesion ability. The hydroxyl, carboxyl or amine groups in its structure play a key role in bioadhesion. At the same time, the electrostatic repulsion between the positively charged ovalbumin and the negatively charged sodium carboxymethyl cellulose will also enhance the transport across the mucus layer. In addition, Fe 3+ - Due to its adhesive properties, the polyphenol network structure can form a bridge with the macroporous honeycomb carboxymethylated β-glucan to achieve complete encapsulation of the complex beneficial bacteria, which is used to protect and prolong the delivery of beneficial bacteria and promote the growth of beneficial colonic microbiota.

[0023] (3) The present invention uses a variety of beneficial bacteria as the main body, and can provide the benefits of a variety of probiotics at one time, wherein the composite beneficial bacteria can supplement the beneficial bacteria in the intestine; oligofructose as a prebiotic is beneficial to the growth and reproduction of beneficial bacteria in the intestine and intestinal health; selenium-enriched yeast can supplement the selenium element in the body, plant enzymes can improve the antioxidant capacity in the body, curcumin can resist inflammation and protect the liver, lower blood lipids, and promote digestion; American ginseng extract can calm the nerves and improve immunity; gelatinized starch helps eliminate fat and increase the nutritional value of health products. Through the synergistic effect of different nutrients, the prepared health product composition has a good taste and the nutrients are easy to absorb, which can not only improve the balance of intestinal microorganisms, but also significantly improve immunity. DETAILED DESCRIPTION

[0024] The technical scheme of the present invention will be clearly and completely described below by embodiments of the present invention. Obviously, the described embodiments are only a part of embodiments of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those skilled in the art without making creative work premise belong to the scope of protection of the present invention. Unless otherwise stated, the raw materials and reagents used in this application are all commercially available products, or can be prepared by known methods.

[0025] The preparation method of American ginseng extract is as follows: slice the American ginseng raw medicinal material, add 10 times the amount of edible ethanol, soak at 25°C for 1 hour, heat under reflux for 2 hours, and obtain the extract after concentration and spray drying.

[0026] The preparation method of gelatinized starch is as follows: sweet potato starch and deionized water are uniformly mixed in a mass ratio of 1:2, the pH value is adjusted to 6-7 with a 5% sodium carbonate aqueous solution, and the gelatinized starch is heated in a water bath at 90-100° C. until the starch is completely formed into a paste.

[0027] The preparation method of plant enzyme powder is as follows: steam bitter gourd and apple in a mass ratio of 2:3 at 100°C for 10 minutes, cool to 30°C, add to a mixture of brown rice enzyme and soybean enzyme in a mass ratio of 1:1 under sterile conditions, and ferment at 32°C for 30 hours.

[0028] Bifidobacterium longum with strain number bio-67291, Lactobacillus acidophilus with strain number bio-67309, Lactobacillus rhamnosus with strain number bio-67204 and Lactococcus lactis with strain number bio-67102 were all purchased from China Industrial Microbiological Culture Collection Center.

[0029] Example 1

[0030] (1) The preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis were inoculated into the corresponding liquid culture medium at a mass ratio of 3:3:2:2, and cultured continuously for 24 hours at a temperature of 40° C. and a rotation speed of 200 r / min. After being mixed evenly, a composite microbial beneficial bacteria was obtained.

[0031] (2) The number of viable cells in 100 μL is 4×10 10 The composite microbial beneficial bacteria with a concentration of CFU / g were placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of tannic acid with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 12 hours, centrifuged at 5500 r / min for 4 minutes, 10 μL of carboxymethylated β-glucan with a mass concentration of 0.2 mg / mL was added for coating, washed with PBS buffer, and freeze-dried to obtain microcapsule composite beneficial bacteria.

[0032] (3) 8 g of egg albumin and 6 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 2.8 with a 25% hydrochloric acid aqueous solution. 1.5 g of curcumin and 2 g of microencapsulated composite beneficial bacteria were added, and stirred evenly at 25° C. and 350 r / min. Then, 35 g of gelatinized starch, 12 g of plant enzyme powder, 4 g of selenium-enriched yeast powder, 1.5 g of American ginseng extract and 15 g of oligofructose were added, and stirred at 45° C. and 900 r / min for 4 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0033] Example 2

[0034] (1) The preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis were inoculated into the corresponding liquid culture medium at a mass ratio of 3:3:2:2, and cultured continuously for 16 hours at a temperature of 50° C. and a rotation speed of 250 r / min. After being mixed evenly, a composite microbial beneficial bacteria was obtained.

[0035] (2) The number of viable cells in 100 μL is 4×10 10 The composite microbial beneficial bacteria with a concentration of CFU / g were placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of anthocyanins with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 8 hours, centrifuged at 6000 r / min for 3 minutes, 10 μL of carboxymethylated β-glucan with a mass concentration of 0.2 mg / mL was added for coating, washed with PBS buffer, and freeze-dried to obtain microcapsule composite beneficial bacteria.

[0036] (3) 7 g of egg albumin and 5 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 3.6 with a 25% hydrochloric acid aqueous solution. 2.5 g of curcumin and 4 g of microencapsulated composite beneficial bacteria were added, and stirred evenly at 30° C. and 500 r / min. Then, 40 g of gelatinized starch, 8 g of plant enzyme powder, 3 g of selenium-enriched yeast powder, 0.5 g of American ginseng extract and 12 g of oligofructose were added, and stirred at 50° C. and 1000 r / min for 2 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0037] Example 3

[0038] (1) The preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis were inoculated into the corresponding liquid culture medium at a mass ratio of 3:3:2:2, and cultured continuously for 32 hours at a temperature of 30° C. and a rotation speed of 150 r / min. After being mixed evenly, a composite microbial beneficial bacteria was obtained.

[0039] (2) The number of viable cells in 100 μL is 4×10 10 The composite microbial beneficial bacteria with a concentration of CFU / g were placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of lignin polyphenols with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 16 hours, centrifuged at 5000 r / min for 5 minutes, 10 μL of carboxymethylated β-glucan with a mass concentration of 0.2 mg / mL was added for coating, washed with PBS buffer, and freeze-dried to obtain microcapsule composite beneficial bacteria.

[0040] (3) 9 g of egg albumin and 8 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 2.5 with a 25% hydrochloric acid aqueous solution. 1 g of curcumin and 6 g of microencapsulated composite beneficial bacteria were added, and stirred evenly at 20° C. and 300 r / min. Then, 30 g of gelatinized starch, 15 g of plant enzyme powder, 5 g of selenium-enriched yeast powder, 2 g of American ginseng extract and 20 g of oligofructose were added, and stirred at 40° C. and 800 r / min for 5 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0041] Example 4

[0042] (1) The preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis were inoculated into the corresponding liquid culture medium at a mass ratio of 3:3:2:2, and cultured continuously for 18 hours at a temperature of 45° C. and a rotation speed of 180 r / min. After being mixed evenly, a composite microbial beneficial bacteria was obtained.

[0043] (2) The number of viable cells in 100 μL is 4×10 10 The composite microbial beneficial bacteria with a concentration of CFU / g were placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of tannic acid with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 15 hours, centrifuged at 5200 r / min for 3.5 minutes, 10 μL of carboxymethylated β-glucan with a mass concentration of 0.2 mg / mL was added for coating, washed with PBS buffer, and freeze-dried to obtain microcapsule composite beneficial bacteria.

[0044] (3) 8 g of egg albumin and 7 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 3.2 with a 25% hydrochloric acid aqueous solution, 2 g of curcumin and 8 g of microencapsulated composite beneficial bacteria were added, and stirred evenly at 25° C. and 420 r / min, followed by adding 35 g of gelatinized starch, 10 g of plant enzyme powder, 4 g of selenium-enriched yeast powder, 1.5 g of American ginseng extract and 16 g of oligofructose, and stirred at 45° C. and 950 r / min for 5 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0045] Example 5

[0046] (1) The preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis were inoculated into the corresponding liquid culture medium at a mass ratio of 3:3:2:2, and cultured continuously for 18 hours at a temperature of 35° C. and a rotation speed of 220 r / min. After being mixed evenly, a composite microbial beneficial bacteria was obtained.

[0047] (2) The number of viable cells in 100 μL is 4×10 10 The composite microbial beneficial bacteria with a concentration of CFU / g were placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of anthocyanin with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 10 hours, centrifuged at 5500 r / min for 5 minutes, 10 μL of carboxymethylated β-glucan with a mass concentration of 0.2 mg / mL was added for coating, washed with PBS buffer, and freeze-dried to obtain microcapsule composite beneficial bacteria.

[0048] (3) 9 g of egg albumin and 5 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 3.0 with a 25% hydrochloric acid aqueous solution. 1.5 g of curcumin and 10 g of microcapsule composite beneficial bacteria were added, and stirred evenly at 30° C. and 420 r / min. Then, 32 g of gelatinized starch, 14 g of plant enzyme powder, 5 g of selenium-enriched yeast powder, 1 g of American ginseng extract and 15 g of oligofructose were added, and stirred at 50° C. and 1000 r / min for 3 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0049] Comparative Example 1

[0050] (1) The number of viable cells in 100 μL is 4×10 10CFU / g of the composite microbial beneficial bacteria (prepared in Example 1) was placed in a liquid culture medium, cultured at a constant temperature of 37°C overnight, resuspended in PBS buffer to obtain a composite microbial beneficial bacteria suspension, 20 μL of tannic acid with a mass concentration of 50 mg / mL and 20 μL of FeCl3·6H2O with a mass concentration of 15 mg / mL were added, reacted at room temperature for 12 h, centrifuged at 5500 r / min for 4 min, washed with PBS buffer, and freeze-dried to obtain encapsulated composite beneficial bacteria.

[0051] (2) 8 g of egg albumin and 6 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 2.8 with a 25% hydrochloric acid aqueous solution. 1.5 g of curcumin and 2 g of microencapsulated composite beneficial bacteria were added, and stirred evenly at 25° C. and 350 r / min. Then, 35 g of gelatinized starch, 12 g of plant enzyme powder, 4 g of selenium-enriched yeast powder, 1.5 g of American ginseng extract and 15 g of oligofructose were added, and stirred at 45° C. and 900 r / min for 4 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0052] Comparative Example 2

[0053] 8 g of egg albumin and 6 g of sodium carboxymethyl cellulose were added to deionized water, stirred to dissolve, and the pH was adjusted to 2.8 with a 25% hydrochloric acid aqueous solution. 1.5 g of curcumin and 2 g of composite microbial beneficial bacteria (prepared by Example 1) were added, and stirred evenly at 25° C. and 350 r / min. Then, 35 g of gelatinized starch, 12 g of plant enzyme powder, 4 g of selenium-enriched yeast powder, 1.5 g of American ginseng extract and 15 g of oligofructose were added, and stirred at 45° C. and 900 r / min for 4 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0054] Comparative Example 3

[0055] The pH of deionized water was adjusted to 2.8 with a 25% hydrochloric acid aqueous solution, 1.5 g of curcumin and 2 g of composite microbial beneficial bacteria (prepared in Example 1) were added, and stirred evenly at 25° C. and 350 r / min. Then, 35 g of gelatinized starch, 12 g of plant enzyme powder, 4 g of selenium-enriched yeast powder, 1.5 g of American ginseng extract and 15 g of oligofructose were added, and stirred at 45° C. and 900 r / min for 4 h. The mixture was freeze-dried and pressed into tablets to obtain a health care composition containing microbial beneficial bacteria.

[0056] Preparation of simulated artificial gastric fluid: 0.35 g of pepsin was dissolved in 100 mL of 0.2% NaCl, and the pH value was adjusted to 2 with 1 mol / L HCl.

[0057] Preparation of simulated artificial intestinal fluid: 0.1 g trypsin and 0.5 g ox bile salt were dissolved in 100 mL of 0.5% NaCl, and the pH value was adjusted to 8 with 1 mol / L NaOH.

[0058] Stability test of health care composition: the prepared health care composition containing beneficial microorganisms was placed in simulated artificial gastric juice, subjected to oscillation disintegration treatment at a speed of 120 r / min for 60 min, the sample was quantitatively diluted to an appropriate amount, live bacteria were counted by pouring method, and the survival rate was calculated.

[0059] The prepared health care composition containing beneficial microorganisms was evenly mixed with simulated artificial intestinal fluid at a ratio of 1:9, and disintegrated at a speed of 200 r / min for 60 min in a constant temperature shaking box at 37°C. The live bacteria were counted by the coating plate method to calculate the survival rate.

[0060] Table 1 Stability performance test

[0061]

[0062] Beneficial bacteria must have the ability to resist the low pH value and high bile salt concentration environment of gastric acid in the human body in order to play a unique probiotic function in the human body; from the test results in the above table, it can be seen that with the increase of the content of microcapsule composite beneficial bacteria, the stability of the health care composition containing microbial beneficial bacteria in simulated artificial gastric juice and simulated artificial intestinal juice gradually increases, indicating that the coating of composite beneficial bacteria can protect the beneficial bacteria under acidic and alkaline conditions and play a probiotic role; this is because on the one hand, carboxymethylated β-glucan has a macroporous honeycomb structure, which can more effectively capture microbial beneficial bacteria, and at the same time, phenolic substances have more hydroxyl groups and higher adhesion ability, which can adsorb carboxymethylated β-glucan to Fe through hydrogen bonding. 3+ -The surface of the polyphenol network structure can achieve the purpose of encapsulating the beneficial bacteria of the complex; on the other hand, ovalbumin, as the most abundant phospholipid-containing glycoprotein in egg white, has good nutritional value and functional properties, and can improve the taste of food; sodium carboxymethyl cellulose is an anionic polysaccharide with good gelation effect, which can produce electrostatic adsorption with the positively charged ovalbumin, effectively encapsulate the microencapsulated composite beneficial bacteria and curcumin, and constitute a protein-polysaccharide coated nanosystem, thereby effectively encapsulating the composite beneficial bacteria and curcumin. Therefore, after multi-layer encapsulation, the composite beneficial bacteria can stably exist in the environment of strong acid and high concentration of bile salts in the gastrointestinal tract, and better play its functional role.

[0063] Adhesion performance test: Caco-2 cells with a preservation number of ZKCC-X1665 were taken for recovery and subculture. The cells grew adherently and were tested after about 5 subcultures. The Caco-2 cells were inoculated into a 6-well plate, the culture medium was replaced every other day, and they grew to a monolayer. The cells were washed with PBS buffer, and 2 mL of a health care composition containing microbial beneficial bacteria was added to the cell well plate. The cells were incubated at 37°C and 5% CO2 for 2 h. The cells were digested with 250 μL of 0.25% trypsin-EDTA for 5 min and collected in a sterile tube for viable bacteria counting. The adhesion number (CFU / cell) = the number of beneficial bacteria adhered to each well / the total number of cells in each well.

[0064] Table 2 Adhesion performance test

[0065] Adhesion number (CFU / cell) Example 1 32.76 Example 2 35.84 Example 3 39.12 Example 4 41.36 Example 5 42.05 Comparative Example 1 30.13 Comparative Example 2 25.32 Comparative Example 3 14.58

[0066] Caco-2 cells are a type of intestinal epithelial cells that are similar in structure and function to differentiated human intestinal epithelial cells. Adhesion to epithelial cells is a prerequisite for the colonization of beneficial bacteria. Adherent bacteria can prolong their residence time in the intestine and improve the impact on host health by changing the local microbial flora or regulating immune responses. The test results in the above table show that with the increase in the content of microcapsule composite beneficial bacteria, the adhesion ability of the health care composition containing microbial beneficial bacteria is enhanced. This is because the protein-polysaccharide coated nanosystem in the outer layer has a high hydrophilicity and adhesion ability, and the hydroxyl, carboxyl or amine group in its structure plays a key role in biological adhesion. At the same time, the electrostatic repulsion between the positively charged ovalbumin and the negatively charged sodium carboxymethyl cellulose will also enhance the transport across the mucus layer. In addition, Fe 3+ - Due to its adhesive properties, the polyphenol network structure can form a bridge with the macroporous honeycomb carboxymethylated β-glucan to achieve complete encapsulation of the complex beneficial bacteria, which is used to protect and prolong the delivery of beneficial bacteria and promote the growth of beneficial colonic microbiota.

[0067] The above-described embodiments provide a detailed description of the technical solutions and beneficial effects of the present invention. It should be understood that the above description is only a specific embodiment of the present invention and is not intended to limit the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected.

Claims

1. A method for preparing a health-care composition containing beneficial microorganisms, characterized in that: The health care composition comprises the following components in parts by weight: 30-40 parts of gelatinized starch, 8-15 parts of plant enzyme powder, 2-10 parts of microcapsule composite beneficial bacteria, 3-5 parts of selenium-enriched yeast powder, 0.5-2 parts of American ginseng extract, 12-20 parts of oligofructose, 1-2.5 parts of curcumin, 7-9 parts of ovalbumin, and 5-8 parts of sodium carboxymethyl cellulose; The preparation method of the health care composition containing beneficial microorganisms is as follows: adding egg white protein and sodium carboxymethyl cellulose to deionized water, stirring and dissolving, adjusting the pH to 2.5-3.6 with a hydrochloric acid aqueous solution, adding curcumin and microcapsule composite beneficial bacteria, stirring evenly at 20-30° C. and 300-500 r / min, then adding gelatinized starch, plant enzyme powder, selenium-enriched yeast powder, American ginseng extract and oligofructose, mixing and heating, stirring evenly, freeze-drying, and pressing into tablets to obtain the health care composition containing beneficial microorganisms; The preparation method of the American ginseng extract is as follows: slice the American ginseng raw medicinal material, add 10 times the amount of edible ethanol, soak at 25°C for 1 hour, heat and reflux for 2 hours, and concentrate and spray-dry the extract to obtain the extract.

2. The method for preparing the health-care composition containing beneficial microorganisms according to claim 1, characterized in that: The preparation method of the gelatinized starch is as follows: sweet potato starch and deionized water are uniformly mixed in a mass ratio of 1:2, the pH value is adjusted to 6-7 with a 5% sodium carbonate aqueous solution, and the gelatinized starch is heated in a water bath at 90-100° C. until the starch is completely formed into a paste.

3. The method for preparing the health-care composition containing beneficial microorganisms according to claim 1, characterized in that: The preparation method of the plant enzyme powder is as follows: steam bitter melon and apple at a mass ratio of 2:3 at 100° C. for 10 minutes, cool to 30° C., add to a mixture of brown rice enzyme and soybean enzyme at a mass ratio of 1:1 under sterile conditions, and ferment at 32° C. for 30 hours to obtain the powder.

4. The method for preparing the health-care composition containing beneficial microorganisms according to claim 1, characterized in that: The mixing heating temperature is 40-50° C., the stirring speed is 800-1000 r / min, and the stirring time is 2-5 h.

5. The method for preparing the health-care composition containing beneficial microorganisms according to claim 1, characterized in that: The preparation method of the microcapsule composite beneficial bacteria is as follows: Step (1), inoculating the preserved freeze-dried bacterial powders of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis into corresponding liquid culture medium, culturing continuously for 16-32 hours at a temperature of 30-50° C. and a rotation speed of 150-250 r / min, and mixing evenly to obtain composite microbial beneficial bacteria; Step (2), placing the composite microbial beneficial bacteria in a liquid culture medium, culturing at a constant temperature of 37° C. overnight, resuspending in a PBS buffer to obtain a composite microbial beneficial bacteria suspension, adding a polyphenol compound and FeCl3·6H2O, reacting at room temperature for 8-16 hours, high-speed centrifuging at 5000-6000 r / min for 3-5 minutes, adding carboxymethylated β-glucan for coating, washing with a PBS buffer, and freeze-drying to obtain microcapsule composite beneficial bacteria.

6. The method for preparing the health-care composition containing beneficial microorganisms according to claim 5, characterized in that: In the step (1), the mass ratio of Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus rhamnosus and Lactococcus lactis is 3:3:2:

2.

7. The method for preparing the health-care composition containing beneficial microorganisms according to claim 5, characterized in that: The polyphenol compound in step (2) is any one of tannic acid, anthocyanin or lignin polyphenol.

8. The method for preparing the health-care composition containing beneficial microorganisms according to claim 5, characterized in that: The viable count of the composite microbial beneficial bacteria suspension in step (2) is 4×10 10 CFU / g; the volume ratio of the composite microbial beneficial bacteria suspension, the polyphenol compound solution, FeCl3·6H2O, and the carboxymethylated β-glucan is 100:20:20:

10.

9. The method for preparing the health-care composition containing beneficial microorganisms according to claim 5, characterized in that: In the step (2), the mass concentrations of the polyphenol compound, FeCl3·6H2O, and carboxymethylated β-glucan are 50 mg / mL, 15 mg / mL, and 0.2 mg / mL, respectively.

10. A health care composition containing beneficial microorganisms, characterized in that: Obtained by the preparation method according to any one of claims 1 to 9.

Images