CN111345473A

CN111345473A - Probiotics composition containing yolk antibody IgY and application preparation

Info

Publication number: CN111345473A
Application number: CN202010174065.3A
Authority: CN
Inventors: 章真; 王宇; 安敏; 王文
Original assignee: Zhuhai Huamin Pharmaceutical Technology Co ltd
Current assignee: Zhuhai Huamin Pharmaceutical Technology Co ltd
Priority date: 2020-03-13
Filing date: 2020-03-13
Publication date: 2020-06-30

Abstract

The invention discloses a probiotic composition containing yolk antibody IgY, which comprises the following components in parts by weight: 5 to 10 parts of yolk extract dry powder containing yolk antibody IgY, 500 to 1000 parts of probiotic powder and 500 to 1000 parts of probiotic protective agent. The invention is suitable for people of different age groups, adopts the optimally combined probiotics, and has the effects of inhibiting harmful pathogenic bacteria, conditioning intestinal tracts, preventing diarrhea and flatulence, improving the immunity of organisms and the like; and the probiotics protective agent is matched to be adopted, so that the probiotics can be protected, and can be successfully planted in the intestinal tract, so that the probiotics can be rapidly proliferated in the intestinal tract, the anti-flatulence effect is better by matching with the design of probiotics, the helicobacter pylori can be rapidly eliminated or the proliferation of the helicobacter pylori can be inhibited by utilizing the targeting property of the egg yolk antibody, and the proliferation of the probiotics is utilized to occupy the space, so that the microenvironment of the intestinal tract is improved, and the immunity of the organism is improved.

Description

Probiotics composition containing yolk antibody IgY and application preparation

Technical Field

The invention relates to the technical field of food, in particular to a probiotic composition containing yolk antibody IgY and an application preparation.

Background

With the improvement of living standard, people pay more and more attention to their health conditions. Immunity is the body's own defense mechanism, and is the body's ability to recognize and destroy any foreign body (virus, bacteria, etc.) that invades from the outside, to treat aged, damaged, dead, denatured self cells, and to recognize and treat mutant cells and virus-infected cells in the body. The body with low immunity is easy to cause the infection of bacteria, virus, fungi and the like, so as to generate disease symptoms such as weak constitution, malnutrition, listlessness, fatigue, weakness, appetite reduction, sleep disorder and the like, which not only aggravates the consumption of the body, but also can recover for a long time when the body is ill every time, and can induce serious diseases in the past. In addition, changes in diet, use of antibiotic drugs, changes in age, intestinal immune dysfunction and the like also affect the health of a human body, so that intestinal flora disorder is caused, sensitive intestinal bacteria are inhibited, and uninhibited bacteria are propagated organically, thereby causing flora imbalance and causing constipation symptoms of the human body.

Probiotics (probiotic) is a kind of active microorganism beneficial to a host, and is a general term for active beneficial microorganisms which are planted in the intestinal tract of a human body and can produce exact health efficacy so as to improve the microbial ecological balance of the host and play beneficial roles. The beneficial bacteria or fungi in human bodies and animal bodies are mainly as follows: clostridium butyricum, lactobacillus, bifidobacterium, actinomycete, saccharomycete, etc. The products with the strongest research functions in the world at present mainly comprise the composite active probiotics consisting of various microorganisms, and are widely applied to the fields of bioengineering, industry and agriculture, food safety and life health.

The probiotics mainly comprise lactobacillus and bifidobacterium, wherein the lactobacillus can promote the secretion of digestive enzyme and intestinal peristalsis, remove intestinal garbage, and simultaneously has good effects of quickly regulating intestinal functions and intestinal flora of patients with gastrointestinal dysfunction caused by long-term oral administration of antibiotics; the Bacillus bifidus can produce lactic acid and acetic acid after fermentation in human intestine, and has effects of increasing utilization rate of calcium, phosphorus and iron, and promoting absorption of iron and vitamin D. The bifidobacterium ferments lactose to produce galactose, which is a component forming cerebroside in the cranial nerve system and has close relation with the rapid growth of the postnatal brain of the infant. The Bifidobacterium can produce nutrients essential to human body such as vitamin B1, B2, B6, B12, alanine, valine, aspartic acid and threonine, has nutrition effect on human body, and can inhibit growth of putrefying bacteria and reduce production of harmful substances such as ammonia, hydrogen sulfide and indole in metabolite. Hundreds of probiotic health products have been developed abroad, including: yogurt, yoghurt, sour soybean milk containing probiotics, oral liquid, tablet, capsule, powder, bacteriostatic spray containing various probiotics, etc. However, the domestic awareness of probiotics is not very strong, so that the product line application of probiotics is not very wide, and the survival rate is a technical problem.

Helicobacter Pylori (HP) is a gram-negative pathogenic bacterium with the highest infection rate among global people, the infection rate of the bacterium in China exceeds 50%, the number of people is over eight hundred million, and not less than 9000 million people need to be treated every year. Helicobacter pylori infection is a major cause of gastritis, gastric ulcer, duodenal ulcer and gastric cancer. Helicobacter pylori is identified as a carcinogen by the world health organization, and more than 95% of patients with gastric cancer death are infected with helicobacter pylori. China is a country with high incidence of gastric cancer, and the stomach cancer patients are newly increased every year and half of the patients died due to the gastric cancer come from China every year. Prevention and treatment of helicobacter pylori infection is the key to reducing the incidence of gastric cancer.

HP-IgY is prepared by injecting helicobacter pylori as antigen into laying hen to obtain helicobacter pylori yolk antibody from yolk. A large number of experimental researches show that the HP-IgY can specifically eliminate helicobacter pylori and has obvious anti-inflammatory effect on inflammatory reactions such as gastritis, gastric ulcer, duodenal ulcer and the like.

However, there is no precedent for combining and using the anti-helicobacter pylori yolk antibody with the probiotics. The composition is used for remarkably improving the quality of intestinal flora and specifically and targetedly eliminating harmful bacteria such as helicobacter pylori and the like.

Disclosure of Invention

In view of the above-mentioned technical deficiencies, it is an object of the present invention to provide a probiotic composition based on egg yolk antibodies.

In order to solve the technical problems, the invention adopts the following technical scheme:

the invention provides a probiotic composition containing yolk antibody IgY, which comprises the following components in parts by weight: 5 to 10 parts of yolk extract dry powder containing yolk antibody, 500 to 1000 parts of probiotic powder and 500 to 1000 parts of probiotic protective agent; wherein the probiotic bacteria is one or more of Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus.

Preferably, the yolk extract dry powder containing the yolk antibody IgY is obtained by obtaining immunized eggs, extracting by a water dilution method, purifying by a sodium chloride salting-out method, purifying by thiophilic affinity chromatography and freeze-drying.

Preferably, the weight ratio of the yolk antibody in the yolk extract dry powder containing the yolk antibody IgY is 5-15%.

Preferably, in the probiotics, the mass ratio of lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus casei, bifidobacterium lactis, bifidobacterium adolescentis, bifidobacterium longum and lactobacillus rhamnosus is 1:1:1:1:1:1: 1.

Preferably, the total bacteria content of the probiotic powder is more than 100 hundred million cfu/g.

Preferably, the probiotic protectant comprises a water-soluble dietary fiber.

Preferably, the water-soluble dietary fiber comprises pectin-based water-soluble dietary fiber.

Preferably, the water-soluble dietary fibre comprises inulin, fructo-oligosaccharide or malto-oligosaccharide.

Preferably, the pectin-based water-soluble dietary fiber comprises at least one of apple pectin, shaddock peel pectin, blueberry pectin and pineapple pectin.

The invention also provides granules and tablets prepared from the probiotic composition.

The invention has the beneficial effects that: the probiotic is suitable for people of different ages, and has the effects of inhibiting harmful pathogenic bacteria, conditioning intestinal tract, preventing diarrhea and flatulence, improving immunity and the like by adopting optimally combined probiotics; on the other hand, the probiotics protective agent is matched to be adopted, so that the probiotics can be protected, and can be planted in the intestinal tract smoothly, so that the probiotics can be proliferated in the intestinal tract rapidly, the anti-flatulence effect is good by matching with the design of probiotics, the helicobacter pylori can be eliminated rapidly or the proliferation of the helicobacter pylori can be inhibited by utilizing the targeting property of the egg yolk antibody, the proliferation of the probiotics is utilized to occupy the space, the microenvironment of the intestinal tract is improved, and the immunity of the organism is improved.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

FIG. 1 is a schematic flow chart diagram of one embodiment of the present invention;

fig. 2 is a statistical analysis chart of the effect evaluation after eating.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

In one embodiment, as shown in fig. 1, a probiotic composition containing yolk antibody IgY comprises the following components in parts by weight: 5-10 parts of yolk extract dry powder containing yolk antibody IgY, 500-1000 parts of probiotic powder and 500-1000 parts of probiotic protective agent; wherein the probiotic powder comprises but is not limited to lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus casei, bifidobacterium lactis, bifidobacterium adolescentis, bifidobacterium longum, lactobacillus rhamnosus and the like, and the total bacteria content is more than 100 hundred million cfu/g. The parts, i.e., parts by mass, are understood to mean grams, milligrams, kilograms, pounds, tons, and the like.

In one embodiment, the probiotic composition is a granule; can be taken orally, or taken after being infused with warm water at a temperature lower than 37 deg.C or taken after being added with water. In one embodiment, the probiotic composition preparation is a tablet, and can be taken by mouth or swallowed. Therefore, the probiotic combined preparation is very convenient to take, and the probiotic combined preparation can be conveniently taken in both home life and journey.

In one embodiment, the probiotic combination preparation comprises probiotic powder, and the total bacteria content of the probiotic powder is more than 100 hundred million cfu/g. CFU/g refers to the total number of probiotics contained per gram of sample, and Colony Forming Units (CFU) refers to the number of viable bacteria per unit volume. In the case of viable bacteria culture counting, a colony formed by growing and propagating a single cell or a plurality of aggregated cells on a solid medium is called a colony forming unit, and the number of viable bacteria is expressed. The colony forming units are metered in a different manner than normal counts and the CFU counts only live bacteria, in order to ensure a sufficient probiotic count and its proliferation rate.

The lactobacillus is one of normal flora of human body, has high intestinal adhesion rate and strong planting capacity, has the important physiological health-care functions of efficiently reducing cholesterol, promoting cell division, conditioning intestinal tract, preventing decayed teeth, improving immunity of organism, resisting cancer and the like, and contains gram-positive anaerobic bacteria without plasmids; lactose is not available, but monosaccharides are metabolized; can grow well under anaerobic condition in the presence of CO₂Can grow in the presence of the catalyst. Can tolerate the digestive tract environment of animals, can be planted in the intestinal tracts of human beings and animals, and has the effects of regulating intestinal flora, preventing and treating diarrhea, removing toxins, improving the immunity of organisms and the like. Has good effect of relieving allergic constitution. Bifidobacterium lactis which has excellent acid bile tolerance, contains bile salt hydrolase, has strong adhesiveness, and can survive in gastric acid and bile after being eaten by a human body, thus being viable through the gastrointestinal tract and maintaining healthy gastrointestinal flora, contributing to improvement of intestinal function, preventing diarrhea, and reducing side effects of antibiotic treatment and improving body immunity, such as antibiotic-associated diarrhea; it also helps to improve the resistance of the body to common respiratory tract infection and reduce the incidence of acute respiratory tract infection. Streptococcus thermophilus (Streptococcus thermophilus) is generally considered to reach only the upper half of the small intestine, unlike bifidobacteria which go to the large intestine and produce lactase, thereby helping lactose intolerant people digest lactose.

In one embodiment, the probiotic combined preparation comprises 500-1000 parts of probiotic protecting agent; the probiotic protecting agent is used for protecting the survival rate of probiotics when the probiotics pass through the stomach esophagus.

In one embodiment, the probiotic protectant includes a water-soluble dietary fiber. Dietary fiber is a carbohydrate that is not digestible by the human body and is divided into two major classes, water-insoluble and water-soluble fiber. The water-soluble dietary fiber can be dissolved in water, can absorb water to swell, and can be fermented by microorganisms in the large intestine, the water-soluble dietary fiber is usually present in plant cell sap and intercellular substance, and mainly contains pectin, plant gum, viscose, etc., the water-soluble dietary fiber is a non-nutrient component in food, but is beneficial to human health, and the substances can stimulate intestinal tract peristalsis, facilitate feces discharge, and can prevent constipation, rectal cancer, hemorrhoid and lower limb varicosis; can be used for preventing cardiovascular diseases such as atherosclerosis and coronary heart disease; preventing the formation of gallstone. It can produce satiety, is beneficial to obesity patients, and can be used as weight reducing food. Improving sugar tolerance, regulating blood sugar level of diabetes patients, and can be used as food for diabetes patients; improving intestinal flora, and preventing intestinal cancer and appendicitis.

The preparation method of the probiotic combined preparation comprises the following steps of part or all of the steps.

And (3) drying: respectively or completely drying the other components except the probiotic powder or respectively or completely drying the other components except the component to be protected; in one embodiment, greenhouse drying, hot air drying or thermal spray drying is used to reduce the moisture content of each component, in one embodiment, the moisture content of each component is reduced to 5% or less; in one embodiment, the moisture content of each component is reduced to 4% or less; in one embodiment, the moisture content of each component is reduced to 3.5% or less; in one embodiment, the moisture content of each component is reduced to 3% or less.

And (3) cooling: the remaining components are cooled to room temperature, which is generally not higher than 35 ℃.

Mixing: adding probiotic powder into the rest components or adding the component to be protected into the rest components, and mixing uniformly; in one embodiment, a screw mixer is used for mixing. In one embodiment, the materials are mixed uniformly by a spiral mixer in a low-temperature environment. In one embodiment, the low temperature environment is less than 10 degrees celsius or less, which helps protect the probiotic. It can be understood that the mixing time is set according to the material quantity, and can be flexibly adjusted according to experience; typically, the mixing time should be greater than 10 minutes.

Subpackaging: packaging in bags and sealing; in one embodiment, the package is sealed according to the specification of 1g, 2g or 5 g; in one embodiment, the package is sealed by vacuum nitrogen filling.

Further, prior to the mixing step, the preparation method further comprises an embedding step: at least part of milk powder, at least part of cocoa powder and a mixing agent are uniformly mixed and embedded with the probiotic powder to form a large amount of probiotic powder embedded bodies, so that the probiotic powder embedded bodies have the outer layers of the milk powder and the cocoa powder which are uniformly mixed. In one embodiment, the probiotic powder is embedded in a large amount of probiotic powder embedded bodies by uniformly mixing milk powder 5-12 times the mass ratio of the probiotic powder, cocoa powder 1-2 times the mass ratio of the probiotic powder and a mixing agent 1-6 times the mass ratio of the probiotic powder to obtain a mixture, and mixing the mixture with the probiotic powder. Further, the mixture and the probiotic powder are mixed under a low-temperature environment of less than 10 ℃ so that the mixture embeds the probiotic powder to form a large amount of probiotic powder embedding bodies. Further, after the embedding step and before the mixing step, the preparation method further comprises a re-drying step: drying the probiotic powder embedded body; in one embodiment, greenhouse drying, hot air drying or thermal spray drying is used.

The probiotic combination formulation of the present invention is further exemplified below in connection with the experimental situation.

Example 1:

the formulation of this example is by weight

The preparation method comprises the following steps:

1. preparation of the starting materials

(1) Preparation of yolk antibody:

p1 immune egg collection: immunizing laying hens by using specific helicobacter pylori antigens, collecting eggs after immunization, and preserving;

p2 yolk antibody extraction by water dilution: taking the immune eggs collected in the step P1, removing egg white under aseptic condition, collecting egg yolk, adding ultrapure water, magnetically stirring, adjusting the pH to 5.0-5.2, continuously magnetically stirring, standing at-20 to-80 ℃ for overnight solidification, unfreezing for 1-4 h at 4 ℃ after completion, then completely unfreezing at room temperature, centrifugally collecting supernatant, filtering through sterilized filter paper, and collecting filtrate;

p3 sodium chloride salting-out purification: adding solid sodium chloride into the filtrate collected in the step P2 under a stirring state, uniformly stirring, adjusting the pH value to 4.0, magnetically stirring, centrifuging, collecting the precipitate and drying in the air;

p4 thiophilic affinity chromatography purification: dissolving the precipitate collected in the step P3 with a buffer solution, filtering with a membrane, then loading and eluting, collecting the eluent, centrifuging, desalting and concentrating, and then filtering with a membrane to obtain the filtrate of the yolk antibody against helicobacter pylori.

(2) Preparing probiotic powder:

s1: seed activation: inoculating the strain kept by liquid nitrogen on a slant for activation culture; culturing in 38 ° incubator for 2 days;

s2, preparing a seed culture medium: 10g of peptone, 5g of glucose, 5g of yeast powder, 2g of inulin, 1g of fructose, 1-8 g of agar (for an alternative solid culture medium), 0.1g of dipotassium hydrogen phosphate, 0.1g of calcium gluconate, 0.05g of zinc gluconate, 0.1g of ferric citrate, 0.1g of sodium chloride and 1000mL of water;

s3: and (3) sterilizing a culture medium: sterilizing at 121 deg.C for 20 min, and cooling;

s4, inoculation: picking the activated bacterial colony into a liquid culture medium in a sterile operation table, and culturing for 2 days at the temperature of 38 ℃ on a shaking table at 100 rpm;

s5: seed amplification culture: the culture solution obtained in S4 was mixed in the following ratio of 1: 10 in a larger volume flask;

s6: anaerobic fermentation: the culture solution obtained in S5 is inoculated into a fermentation tank for anaerobic fermentation. The culture temperature is 38-44 ℃, and the stirring speed is 50-100 r/min;

s7, harvesting: the culture broth obtained in S6 was centrifuged at low speed in a centrifuge at a centrifugal force of 2000g for 10 minutes. And is rinsed with pure water and then centrifuged once;

s8, spray drying: and (S7) adding protective agent skimmed milk powder into the bacterial sludge obtained in the step (A), mixing uniformly, and adjusting the solid content to 20% by using pure water. Then pumping the mixture into a spray drying tower (inlet air temperature is 140 ℃, outlet air temperature is 80 ℃, and flow rate is 1L/h) by a peristaltic pump to obtain the probiotic powder

(3) The water-soluble dietary fiber is a commercial product;

2. weighing and preparing materials

All materials are weighed according to the formula amount, and the materials except the probiotics need to be dried until the moisture is less than 1%.

3. Mixing

And (3) placing all the raw and auxiliary materials in a mixer, and mixing until the color and texture of the mixed material are uniform.

4. Pressing sheet (alternative)

Tabletting with a tabletting machine, and periodically checking for tablet weight differences.

Example 2:

the formulation of this example is by weight

The rest of the preparation method is the same as that of the example 1, and the specific steps are omitted here.

Example 3:

the formulation of this example is by weight

Example 4:

the formulation of this example is by weight

Example 5:

the formulation of this example is by weight

Comparative example 1:

the formulation of this example is by weight

Comparative example 2:

the formulation of this example is by weight

The experimental verification method comprises the following steps:

1. simulated gastric juice tolerance test in vitro probiotic composition was passed through the stomach and the tolerance of the composition was evaluated in terms of the number of survivors after its passage each composition was added to prepared sterile simulated gastric juice (5 × 10) in a ratio of 1: 2⁹cfu/mL), shaking, mixing, placing in a constant temperature incubator at 37 ℃, and taking 1mL of culture solution to perform gradient dilution viable bacteria counting for 1h, 2h and 3h respectively. Preparing artificial gastric juice: diluting with 10% hydrochloric acid 16.4mL with deionized water to pH 2.0, adding pepsin (1g/100mL), dissolving completely, filtering with 0.22 μm microporous membrane, and sterilizing.

2. Evaluation of post-eating Effect

Test volunteers selection criteria: the patients with age of 25-60 years, no other basic diseases, mild heartburn, acid regurgitation and halitosis.

100 of the enrolled volunteers were randomly selected to participate in the experiment. 0.5 g of the prepared composition was administered once a day, morning and evening, and orally. The life regularity and habits of the experimenters remained unchanged during the test period. The observation days were 30 days. After the test is finished, the score of the trial effect is obtained through questionnaire survey. The proportion of the patients with reduced chief complaints and no obvious change is counted and analyzed, and the statistical result is shown in figure 2.

Although the present invention has been described in detail with reference to the above embodiments, those skilled in the art can make modifications and equivalents to the embodiments of the present invention without departing from the spirit and scope of the present invention, which is set forth in the claims of the present application.

Claims

1. A probiotic composition containing yolk antibody IgY comprises the following components in parts by weight: 5-10 parts of yolk extract dry powder containing yolk antibody IgY, 500-1000 parts of probiotic powder and 500-1000 parts of probiotic protective agent; wherein the probiotic bacteria is one or more of Lactobacillus bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium longum, and Lactobacillus rhamnosus.

2. The probiotic composition according to claim 1, wherein the yolk extract dry powder containing the yolk antibody IgY is obtained by obtaining immunized eggs, extracting by water dilution, purifying by sodium chloride salting-out method, purifying by thiophilic affinity chromatography, and freeze-drying.

3. The probiotic composition according to claim 1, wherein the weight ratio of the yolk antibodies in the yolk extract dry powder containing the yolk antibodies IgY is 5-15%.

4. The probiotic composition according to claim 1, wherein the probiotic comprises lactobacillus bulgaricus, streptococcus thermophilus, lactobacillus acidophilus, lactobacillus plantarum, lactobacillus casei, bifidobacterium lactis, bifidobacterium adolescentis, bifidobacterium longum and lactobacillus rhamnosus in a mass ratio of 1:1:1:1:1:1:1: 1.

5. The probiotic composition according to claim 1, characterized in that the total bacterial content of the probiotic powder is greater than 100 hundred million cfu/g.

6. The probiotic composition according to claim 1, characterized in that the probiotic protective agent comprises a water-soluble dietary fiber.

7. The probiotic composition according to claim 6, characterized in that the water-soluble dietary fibers comprise pectin-based water-soluble dietary fibers.

8. The probiotic composition according to claim 6, characterized in that the water-soluble dietary fiber comprises inulin, fructo-oligosaccharide or malto-oligosaccharide.

9. The probiotic composition according to claim 7, wherein the pectin-based water-soluble dietary fiber comprises at least one of apple pectin, grapefruit peel pectin, blueberry pectin, and pineapple pectin.

10. Granules and tablets made from the probiotic composition of any of claims 1-9.