CN110638841B - Probiotics and prebiotics composite preparation, preparation method and application thereof - Google Patents
Probiotics and prebiotics composite preparation, preparation method and application thereof Download PDFInfo
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P5/00—Drugs for disorders of the endocrine system
- A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
- A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention discloses a probiotic and prebiotic composite preparation, a preparation method and application thereof, and belongs to the field of biological medicine. The invention consists of composite probiotics and composite prebiotics, wherein the composite probiotics consist of clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei. The type of the compound probiotics is acquired from intestinal microorganisms of diabetics and healthy people, theoretical research to experimental exploration and result verification, so that the probiotic prebiotic compound preparation which can truly prevent and treat diabetes is obtained; the research results show that the medicine can effectively reduce blood sugar, improve insulin resistance and protect organs.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a probiotic and prebiotic composite preparation, a preparation method and application thereof.
Background
It was counted that about 340 ten thousand people die worldwide in 2004 due to diabetes and its complications. The investigation result of the international diabetes union shows that the global diabetes patients in 2007 are 2.4 hundred million, and the number of patients in 2011 is increased to 3.47 hundred million. The international union predicts that the number of people suffering from diabetes mellitus worldwide will reach 6.29 hundred million for 2045 years. Research reports that China has become a large country of diabetes, and the prevalence rate is continuously increasing. Hyperglycemia is often accompanied by disturbance of blood lipid metabolism, which causes blood viscosity and blood flow velocity reduction, and chronic cardiovascular diseases such as atherosclerosis and thrombosis can be caused by long-term development. Diabetes and complications thereof seriously threaten the physical health of people, and reduce the life quality of people.
Oral hypoglycemic agents are generally used for treating diabetics in clinic, such as sulfonylureas, biguanides and the like. However, long-term application of the treatment method is easy to cause side effects such as diarrhea, dizziness, nausea, inappetence, hypoglycemia, liver function injury and the like, and seriously threatens the health of people. Therefore, the development of a natural and safe novel hypoglycemic agent or adjuvant therapy agent is becoming particularly important.
In recent years, probiotics are becoming research hot spots for preventing and treating diabetes due to the characteristics of safety, availability, low cost, no side effect and the like.
Disclosure of Invention
In order to make up the defects of the prior art, the invention provides a probiotic and prebiotic composite preparation, a preparation method and application thereof.
The technical scheme of the invention is as follows:
a probiotic and prebiotic compound preparation consists of compound probiotics and compound prebiotics, wherein the compound probiotics consist of clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei.
Preferably, in the composite probiotics, the number of live clostridium butyricum is 10 9 ～10 10 CFU/g, the viable count of lactobacillus plantarum is 10 9 ～10 10 CFU/g, viable count of bifidobacterium longum is 10 9 ～10 10 CFU/g, viable count of Lactobacillus paracasei is 10 9 ～10 10 CFU/g。
As a preferable scheme, the composite probiotics consist of 35-55% clostridium butyricum, 20-45% lactobacillus plantarum, 15-30% bifidobacterium longum and 10-25% lactobacillus paracasei according to mass fraction.
Preferably, the complex prebiotic consists of isomaltooligosaccharide and galactooligosaccharide.
Further, in the composite prebiotics, the mass percentage of the isomaltooligosaccharide is 25-85% and the mass percentage of the galactooligosaccharide is 15-75%.
As a preferable scheme, the mass percentage of the composite probiotics is 35% -85%, and the mass percentage of the composite probiotics is 15% -65%.
Most preferably, the mass percentage of the composite probiotics is 72 percent and the mass percentage of the composite probiotics is 28 percent.
The preparation method of the probiotic and prebiotic composite preparation comprises the steps of placing clostridium butyricum powder, lactobacillus plantarum powder, bifidobacterium longum powder and lactobacillus paracasei powder in a sterile tank, uniformly stirring, adding the composite prebiotic, continuously uniformly stirring, and obtaining the probiotic and prebiotic composite preparation.
As a preferred scheme, the preparation method of clostridium butyricum powder, lactobacillus plantarum powder, bifidobacterium longum powder and lactobacillus paracasei powder comprises the following steps:
1) Activating clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei by an MRS culture medium, and then placing the activated clostridium butyricum, lactobacillus paracasei in a fermentation culture medium for high-density culture;
2) Centrifugally separating thalli, adding the thalli into freeze-drying protection liquid containing skimmed milk powder, glutamic acid and vitamin E, mixing, and freeze-drying to obtain corresponding fungus powder.
Preferably, the probiotic and prebiotic composite preparation is powder, tablet or capsule.
The application of the probiotics and prebiotics composite preparation is used for reducing blood sugar and improving insulin resistance.
The related research of human microbiology and various diseases based on high-throughput sequencing provides people with an indication of selecting probiotics on a great basis. In order to reveal the causal relationship between the diabetes patients and the intestinal flora disorder and find a proper probiotic combination drug, the applicant collects a large number of faecal samples of the diabetes patients and healthy people in the early stage, obtains the big data of the intestinal microorganisms of the diabetes patients and healthy people through macro genome sequencing analysis, integrates the big data with the existing intestinal data of the diabetes patients, and constructs the diabetes intestinal flora gene set. Meanwhile, classification of species of intestinal flora of diabetes and healthy people and gene function annotation are completed, and species and functional genes specific to various diabetes and microbiota with remarkably reduced content relative to healthy people are found through comparison. Four specific microorganisms (clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei) are further accurate through a random forest model, model verification is carried out based on independent crowds, and the four microorganism combination changes can be used for well predicting early diabetes.
The prebiotics can provide food for probiotics, can be decomposed and absorbed by beneficial bacteria in intestinal tracts, and promote the growth and propagation of the beneficial bacteria, thereby having beneficial effects on hosts. Based on four mixed probiotics, two probiotics are added, so that continuous energy is further provided for the probiotics, and the novel compound preparation with obvious blood sugar reducing function is provided.
The beneficial effects of the invention are as follows:
1. the probiotics and prebiotics composite preparation provided by the invention is formed by compounding composite probiotics and composite prebiotics; the type of the compound probiotics is acquired from intestinal microorganisms of diabetics and healthy people, theoretical research to experimental exploration and result verification, so that the probiotic prebiotic compound preparation which can truly prevent and treat diabetes is obtained; the research results show that the medicine can effectively reduce blood sugar, improve insulin resistance and protect organs.
2. The probiotics and prebiotics compound preparation provided by the invention is easy to safely have no toxin effect, can be used as a dietary supplement or a nutritional agent, is used for preventing diabetes or is used for assisting in treating diabetes and relieving diabetic complications, and greatly relieves the pain of diabetics. 3. The invention not only can play the role of reducing blood sugar of the probiotics, but also can play other probiotic functions of the probiotics, such as regulating immune function, resisting oxidation, delaying aging and the like, and has important significance on human health.
4. The compound probiotics and the compound prebiotics can effectively inhibit the growth of harmful bacteria, and no preservative is required to be added, so that the risk of side effects caused by the addition of the preservative is reduced.
5. The composite preparation of the invention has the advantages of easily obtained raw materials, simple preparation method, low cost and easy industrialized production.
Detailed Description
Example 1 preparation method of probiotic and prebiotic Complex formulation
1) Preparation of probiotic powder
Preparation of clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei seed liquid:
clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and Lactobacillus paracasei were inoculated into MRS liquid medium at an inoculum size of 4%, respectively, and activated at 37deg.C for 16h. And (3) a small amount of bacterial liquid is dipped by an inoculating loop, three-area line drawing is carried out on the MRS solid culture medium, the culture is carried out for 48 hours at 37 ℃, single bacterial colony is selected to the fermentation culture medium, and the culture is carried out for 16 hours at 37 ℃.
Centrifugally separating thalli, adding the thalli into freeze-drying protection liquid containing 10% of skimmed milk powder, 0.8% of glutamic acid and 1% of vitamin E, and mixing and freeze-drying to obtain corresponding fungus powder. The number of viable bacteria in each bacterial powder is 10 9 -10 10 CFU/g。
Wherein,
the MRS liquid culture medium comprises the following components in percentage by mass: 2.0% of glucose, 0.2% of ammonium citrate, 0.5% of sodium acetate, 0.03% of manganese sulfate, 0.05% of magnesium sulfate, 1.0% of peptone, 1.0% of beef extract, 0.5% of yeast extract, 0.2% of soil temperature and the balance of water; the pH is 6.0-6.2.
The MRS solid culture medium comprises the following components in percentage by mass: 2.0% of glucose, 0.2% of ammonium citrate, 0.5% of sodium acetate, 0.03% of manganese sulfate, 0.05% of magnesium sulfate, 1.0% of peptone, 1.0% of beef extract, 0.5% of yeast extract, 0.2% of soil temperature-80, 1.5% of agar powder and the balance of water; the pH is 6.0-6.2.
The fermentation medium is MRS liquid medium.
2) Proportioning materials
7.2Kg of composite probiotics and 2.8Kg of composite prebiotics.
Wherein the composite probiotics comprise 3.24Kg of clostridium butyricum powder, 1.80Kg of lactobacillus plantarum powder, 1.44Kg of bifidobacterium longum powder and 0.72Kg of lactobacillus paracasei powder.
The composite prebiotics contained 1.82Kg of isomaltooligosaccharide and 0.98Kg of galactooligosaccharide.
3) Mixing
Sequentially adding the above 10 into a sterile stirring tank 9 -10 10 CFU/g Clostridium butyricum powder, 10 9 -10 10 CFU/g Lactobacillus plantarum powder, 10 9 -10 10 CFU/g Bifidobacterium longum powder, 10 9 -10 10 CFU/g lactobacillus paracasei powder, stirring for 20 minutes; and then sequentially adding isomaltooligosaccharide and galactooligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotic composite preparation.
Example 2 preparation of probiotic and prebiotic Complex formulation
1) Proportioning materials
6.5Kg of composite probiotics and 3.5Kg of composite prebiotics.
Wherein the composite probiotics comprise 2.275Kg clostridium butyricum powder, 1.30Kg lactobacillus plantarum powder, 1.95Kg bifidobacterium longum powder and 0.975Kg lactobacillus paracasei powder. The fungus powder adopts the step of the example 1
1) The prepared bacterial powder.
The composite prebiotics contained 1.05Kg of isomaltooligosaccharide and 2.45Kg of galactooligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank 9 -10 10 CFU/g Clostridium butyricum powder, 10 9 -10 10 CFU/g Lactobacillus plantarum powder, 10 9 -10 10 CFU/g Bifidobacterium longum powder, 10 9 -10 10 CFU/g lactobacillus paracasei powder, stirring for 20 minutes; and then sequentially adding isomaltooligosaccharide and galactooligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotic composite preparation.
Example 3 preparation of probiotic and prebiotic Complex formulation
1) Proportioning materials
4.0Kg of composite probiotics and 6.0Kg of composite prebiotics.
Wherein the composite probiotics comprise clostridium butyricum bacterial powder 1.60Kg, lactobacillus plantarum bacterial powder 1.00Kg, bifidobacterium longum bacterial powder 1.00Kg and lactobacillus paracasei bacterial powder 0.40Kg. The bacterial powder prepared in the step 1) of the example 1 is adopted.
The composite prebiotics contained 3.30Kg of isomaltooligosaccharide and 2.70Kg of galactooligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank 9 -10 10 CFU/g Clostridium butyricum powder, 10 9 -10 10 CFU/g Lactobacillus plantarum powder, 10 9 -10 10 CFU/g Bifidobacterium longum powder, 10 9 -10 10 CFU/g lactobacillus paracasei powder, stirring for 20 minutes; and then sequentially adding isomaltooligosaccharide and galactooligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotic composite preparation.
Example 4 preparation of probiotic and prebiotic Complex formulation
1) Proportioning materials
6.0Kg of composite probiotics and 4.0Kg of composite prebiotics.
Wherein the composite probiotics comprise 2.40Kg of clostridium butyricum powder, 1.20Kg of lactobacillus plantarum powder, 1.20Kg of bifidobacterium longum powder and 1.20Kg of lactobacillus paracasei powder. The bacterial powder prepared in the step 1) of the example 1 is adopted.
The composite prebiotics contained 3.00Kg of isomaltooligosaccharide and 1.00Kg of galactooligosaccharide.
2) Mixing
Sequentially adding the above 10 into a sterile stirring tank 9 -10 10 CFU/g Clostridium butyricum powder, 10 9 -10 10 CFU/g Lactobacillus plantarum powder, 10 9 -10 10 CFU/g Bifidobacterium longum powder, 10 9 -10 10 CFU/g lactobacillus paracasei powder, stirring for 20 minutes; and then sequentially adding isomaltooligosaccharide and galactooligosaccharide, and continuously stirring for 10 minutes to obtain the probiotic and prebiotic composite preparation.
Example 5 probiotic and prebiotic Complex formulation
In the ingredients, the composite probiotics account for 80% of the mass of the composite preparation, and the composite probiotics account for 20% of the treatment of the composite preparation. The mass ratio of each bacterial powder in the composite probiotics and the mass ratio of two components in the composite probiotics are the same as in the embodiment 1.
Example 6 probiotic and prebiotic Complex formulation
In the ingredients, the composite probiotics account for 35% of the mass of the composite preparation, and the composite probiotics account for 65% of the treatment of the composite preparation. The mass ratio of each bacterial powder in the composite probiotics and the mass ratio of two components in the composite probiotics are the same as in the embodiment 1.
Example 7 probiotic and prebiotic Complex formulation
The mass ratio of each bacterial powder in the composite probiotics is the same as that of the example 1, and the rest is the same as that of the example 1.
The mass fraction of clostridium butyricum powder in the composite probiotics is 35%, the mass fraction of lactobacillus plantarum powder is 25%, the mass fraction of bifidobacterium longum powder is 25%, and the mass fraction of lactobacillus paracasei powder is 15%.
Example 8 probiotic and prebiotic Complex formulation
The mass ratio of each bacterial powder in the composite probiotics is the same as that of the example 1, and the rest is the same as that of the example 1.
The mass fraction of clostridium butyricum powder in the composite probiotics is 50%, the mass fraction of lactobacillus plantarum powder is 20%, the mass fraction of bifidobacterium longum powder is 20%, and the mass fraction of lactobacillus paracasei powder is 10%.
Example 9 probiotic and prebiotic Complex formulation
The mass ratio of each bacterial powder in the composite probiotics is the same as that of the example 1, and the rest is the same as that of the example 1.
The mass fraction of clostridium butyricum powder in the composite probiotics is 35%, the mass fraction of lactobacillus plantarum powder is 30%, the mass fraction of bifidobacterium longum powder is 25%, and the mass fraction of lactobacillus paracasei powder is 10%.
Example 10 environmental tolerance determination of four probiotic artificial gastrointestinal fluids
(1) Preparation of artificial stomach intestine liquid
A certain amount of gastric protein was dissolved in PBS buffer solution at pH 3.0, and its final concentration was adjusted to 3g/L, thereby obtaining artificial gastric juice.
A certain amount of trypsin was dissolved in PBS buffer solution of pH 8.0, and the final concentration was adjusted to 1g/L, thereby obtaining an artificial intestinal juice.
Both the artificial gastric juice and the artificial intestinal juice were filtered under aseptic conditions with 0.22 μm filters.
(2) Resistance to artificial gastrointestinal fluids
The target strain is inoculated in MRS liquid culture medium with an inoculation amount of 3%, and after three generations of continuous activation at 37 ℃, the bacterial cells are collected by centrifugation and prepared into bacterial suspension for standby. The prepared bacterial suspension is inoculated into artificial gastric juice filtered by a 0.22 mu m filter membrane according to the inoculation amount of 3 percent, cultured at 37 ℃, and 100 mu L of bacterial liquid is respectively taken for dilution and coating at 0, 1, 2 and 3 hours, so as to calculate the survival rate. Then, 1mL of the bacteria-containing artificial gastric juice which is cultured for 3 hours is sucked, 9mL of the bacteria-containing artificial gastric juice which is filtered by a 0.22 mu m filter membrane is added, the bacteria-containing artificial gastric juice is cultured at 37 ℃, and 100 mu L of bacteria solution is respectively sucked at 1, 3, 5 and 8 hours for dilution and coating, and the survival rate is calculated.
The survival rate calculation formula is as follows:
wherein N is t N as the number of colonies after treatment 0 Is the number of untreated colonies.
The environmental tolerance results of the four probiotics artificial gastrointestinal fluids are shown in tables 1-4.
TABLE 1 results of tolerance of Clostridium butyricum to artificial gastrointestinal fluids
TABLE 2 results of tolerance of Lactobacillus plantarum to Artificial gastrointestinal fluids
TABLE 3 results of Bifidobacterium longum tolerance to artificial gastrointestinal fluids
TABLE 4 results of tolerance of Lactobacillus paracasei to Artificial gastrointestinal fluids
As shown in tables 1-4, the survival rate of the four strains in the artificial gastric juice is above 85%, and the survival rate of the four strains in the artificial intestinal juice is above 75%, which shows that the four strains have better tolerance in the artificial simulated gastric intestinal juice, in particular to lactobacillus paracasei.
Example 11 Effect of probiotic and prebiotic Complex formulation on diabetic mice
Grouping and establishing a diabetes model:
360 male C57BL/6J mice are selected for experiment, the age of the mice is about 3 weeks, the mice are randomly divided into 3 groups, and 120 mice in each group: normal, model, and experimental groups. The temperature of the rat room is 22+/-2 ℃, the humidity is 55+/-5%, the day and night alternation is carried out for 12 hours, and the basic feed is suitable for feeding for one week. Starting from week 2, the model group and the experimental group were fed with high-fat and high-sugar feed. After 3 weeks, model and experimental groups were intraperitoneally injected with STZ at a dose of 110mg/kg to construct a diabetes model, while normal groups were injected with the same dose of citric acid buffer as a control. After one week of modeling, the random blood sugar of the mice was measured, and the blood sugar value was higher than 11.1 mmol/L. The experimental group was then perfused with the probiotic and prebiotic complex formulation obtained in example 1, at 9:00 am, and the normal and model groups were perfused with the same dose of saline. The experimental period was 13 weeks in total.
Measurement of blood glucose, insulin: before and after the end of the experiment, the blood glucose values of the mice were measured and recorded. After the mice were fasted overnight without water withdrawal, tail blood was taken and their FBG values were determined. The PBG value of the mice was determined 2 hours after feeding the feed. Insulin was measured after the completion of the experiment, and the results are shown in tables 5, 6 and 7.
TABLE 5 susceptibility of the probiotic and prebiotic composite formulation of example 1 to blood glucose changes before and after experiments in diabetic mice
As can be seen from table 5, the probiotic and prebiotic composite preparation of the present invention can reduce the hyperglycemia level to some extent, and delay the occurrence of diabetes.
TABLE 6 Effect of the probiotic and prebiotic Complex formulation from example 1 on insulin content in diabetic mice
As shown in Table 6, the probiotic and prebiotic composite preparation of the invention can reduce the insulin release amount of diabetic mice and relieve hyperinsulinemia.
TABLE 7 Effect of probiotic and prebiotic Complex formulation from example 1 on organ index in diabetic mice
As shown in Table 7, the probiotic and prebiotic composite preparation can inhibit the increment of organ index of diabetic mice, has a certain protection effect on organs and relieves diabetic complications.
The above description is only for the purpose of illustrating the invention, and it should be understood that the invention is not limited to the above embodiments, but various modifications consistent with the idea of the invention are within the scope of the invention.
Claims (4)
1. The composite preparation of probiotics and prebiotics consists of composite probiotics and composite prebiotics and is characterized in that: the composite probiotics and the composite prebiotics consist of 32.4% clostridium butyricum, 18.0% lactobacillus plantarum, 14.4% bifidobacterium longum, 7.2% lactobacillus paracasei, 18.2% isomaltooligosaccharide and 9.8% galactooligosaccharide according to mass fraction;
in the composite probiotics, the viable count of clostridium butyricum is 10 9 ~10 10 CFU/g, the viable count of lactobacillus plantarum is 10 9 ~10 10 CFU/g, viable count of bifidobacterium longum is 10 9 ~10 10 CFU/g, viable count of Lactobacillus paracasei is 10 9 ~10 10 CFU/g。
2. A method of preparing a probiotic and prebiotic composite formulation according to claim 1, wherein: placing clostridium butyricum powder, lactobacillus plantarum powder, bifidobacterium longum powder and lactobacillus paracasei powder into a sterile tank, uniformly stirring, adding the composite prebiotics, continuously uniformly stirring, and benefiting from the composite preparation of the probiotics and the prebiotics.
3. The method for preparing the probiotic and prebiotic composite preparation according to claim 2, wherein the preparation method of clostridium butyricum powder, lactobacillus plantarum powder, bifidobacterium longum powder and lactobacillus paracasei powder comprises the following steps:
1) Activating clostridium butyricum, lactobacillus plantarum, bifidobacterium longum and lactobacillus paracasei by an MRS culture medium, and then placing the activated clostridium butyricum, lactobacillus paracasei in a fermentation culture medium for high-density culture;
2) Centrifugally separating thalli, adding freeze-drying protection liquid containing skimmed milk powder, glutamic acid and vitamin E, mixing, and freeze-drying to obtain corresponding fungus powder.
4. A method of preparing a probiotic and prebiotic combination formulation according to claim 2 or 3, wherein: the probiotic and prebiotic composite preparation is powder, tablet or capsule.
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