CN113261592A

CN113261592A - Probiotics goat milk powder and preparation method thereof

Info

Publication number: CN113261592A
Application number: CN202110614503.8A
Authority: CN
Inventors: 彭星星
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-06-02
Filing date: 2021-06-02
Publication date: 2021-08-17

Abstract

The invention discloses a probiotic goat milk powder and a preparation method thereof, wherein the probiotic goat milk powder comprises the following raw materials: goat milk, whey protein powder, peanut oil, walnut oil, plant nutrition powder, trehalose, L-carnitine, choline tartrate, compound vitamins, compound minerals, arachidonic acid, docosahexaenoic acid and a probiotic composition. The probiotic goat milk powder disclosed by the invention is rich and comprehensive in nutrition and good in taste, and can supplement nutrient elements for a human body in time, improve the gastrointestinal function of the human body and improve the immunity of the human body.

Description

Probiotics goat milk powder and preparation method thereof

Technical Field

The invention belongs to the technical field of food, and particularly relates to probiotic goat milk powder and a preparation method thereof.

Background

The milk powder is a mixed food made up by using fresh cow milk or goat milk as raw material through the processes of removing water content by adopting freezing or heating process, drying and adding nutrient supplementing components, such as vitamins and minerals, etc.. The goat milk is rich in protein, fat, vitamins, mineral substances and the like, has fat particles which are easy to absorb by human bodies, has high whey protein content and is not easy to cause allergy, is used as a main milk source which is parallel to the milk and the buffalo milk, and is a refined product in the dairy product. Chinese patent CN103651857A discloses an infant formula containing probiotics, its preparation method and its storage method, the infant formula containing probiotics comprises the following raw materials: raw milk, lactose, desalted whey powder, whey protein concentrate, structural grease, prebiotics, infant vitamin premix, infant mineral premix and active probiotic freeze-dried powder, wherein the viable count of the probiotics is stable, the storage time is long, the metabolic activity of total intestinal flora can be effectively stimulated, the immune system is improved, and the intestinal flora is balanced; but the content of the probiotics is not high, and the nutritional formula can be further optimized and improved.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides probiotic goat milk powder and a preparation method thereof.

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

the probiotic goat milk powder comprises the following raw materials: goat milk, whey protein powder, peanut oil, walnut oil, plant nutrition powder, trehalose, L-carnitine, choline tartrate, compound vitamins, compound minerals, arachidonic acid, docosahexaenoic acid and a probiotic composition.

The probiotic goat milk powder is prepared from the following raw materials in parts by weight: 400 parts of goat milk, 30-60 parts of whey protein powder, 1-3 parts of peanut oil, 1-2 parts of walnut oil, 3-5 parts of plant nutrient powder, 0.3-0.5 part of trehalose, 0.05-0.08 part of L-carnitine, 0.03-0.05 part of choline tartrate, 0.1-0.2 part of vitamin complex, 0.1-0.2 part of compound mineral, 0.1-0.3 part of arachidonic acid, 0.1-0.2 part of docosahexaenoic acid and 1-2 parts of probiotic composition.

The compound vitamin is a mixture of L-ascorbic acid, retinol, cholecalciferol, folic acid and D-calcium pantothenate according to the mass ratio of 5 (1-2): (0.5-1): (0.3-0.5): (0.3-0.5).

The compound mineral substance is a mixture of calcium chloride, ferrous sulfate, magnesium sulfate and sodium selenate according to a mass ratio of 15 (1-5) to (1-3) to (0.05-0.1).

The plant nutrition powder is a mixture of burdock powder, cassia seed powder and tuckahoe powder according to a mass ratio of (1-2) to (3).

The probiotic composition consists of 20-60 wt% of bacillus natto freeze-dried powder, 20-40 wt% of lactobacillus johnsonii freeze-dried powder and 20-40 wt% of akkermansia muciniphila freeze-dried powder. Preferably, the probiotic composition consists of 50 wt% of bacillus natto freeze-dried powder, 25 wt% of lactobacillus johnsonii freeze-dried powder and 25 wt% of akkermansia muciniphila freeze-dried powder.

B, bacillus natto: the bacillus natto has the effects of reducing and stabilizing blood sugar, can be applied to the preparation of medicines or preparations for reducing and stabilizing blood sugar, and has obvious effects of promoting the in vivo reduction of blood sugar and stabilizing blood sugar in intestinal expression; meanwhile, the bacillus natto has the functions of obviously losing weight, reducing the level of serum triglyceride, reducing the weight of white fat and reducing the diameter of fat cells; in addition, the bacillus natto enters the intestinal tract of an animal and survives for a long time, so that the immunity of the animal is enhanced for the growth and development of the animal, the number of dominant bacteria in the intestinal tract can be adjusted, the nutrient substances in the intestinal tract are changed, and the constipation is prevented.

Lactobacillus johnsonii: the lactobacillus johnsonii has the effects of improving obesity, reducing body fat and cholesterol, stabilizing intestinal bacteria phase, maintaining intestinal ecology with more bacteria and less bacteria, and further stimulating a mucosa to strengthen an immune system. The lactobacillus johnsonii can stimulate receptors on dendritic cells on the intestinal wall and combine, activate and activate transfer proteins in cells to move into nuclei to release a large amount of cytokines, promote secretion increase of leucocyte and interferon gamma, and inhibit immunoglobulin IgE by stimulating intestinal tract regulation immune response, so that an allergy phenomenon of excessive immune response is improved; meanwhile, the composition has good gastric acid and bile salt tolerance and cell adhesion, can colonize in the intestinal tract of a human body for more than 60 days, and also has good inhibition effect on pathogenic bacteria such as enterococcus faecalis, Escherichia coli, salmonella typhimurium and the like, thereby achieving the effects of cleaning and protecting the intestinal tract.

Akkermansia muciniphila: the akkermansia muciniphila has the function of improving the intestinal barrier. When the intestinal barrier is damaged, the intestinal permeability is increased, and pathogenic microorganisms, endotoxin and the like in the intestinal tract can enter blood or tissues through the intestinal barrier system, so that the body is in a disease state. Therefore, maintaining the intestinal barrier function normally plays a crucial role in the health status of the body. In addition, numerous studies have shown that the occurrence, development and outcome of many diseases (e.g., obesity, diabetes, pancreatitis, fatty liver, inflammatory bowel disease, cancer, etc.) are clinically closely related to intestinal barrier dysfunction. Therefore, intervention treatment for intestinal barrier dysfunction has a very positive effect on blocking disease progression and accelerating disease recovery.

The preparation method of the bacillus natto freeze-dried powder comprises the following steps:

s1, strain activation: marking out the bacillus natto on a solid culture medium A, and culturing for 18-30h at 32-40 ℃ to obtain activated bacillus natto; the preparation method of the solid culture medium A comprises the following steps: uniformly mixing 1-6 wt% of peptone, 1-4 wt% of glucose, 0.2-1 wt% of NaCl, 1-3 wt% of agar powder and the balance of deionized water, and sterilizing at the temperature of 110-;

s2, expanding culture: inoculating activated bacillus natto into a liquid culture medium A, wherein the inoculation amount is 1-5 wt%, and performing shake culture at 34-42 ℃ and 200-; the preparation method of the liquid culture medium A comprises the following steps: 6-14 wt% of skim milk powder, 1-4 wt% of organic inulin, 0.1-2 wt% of potassium dihydrogen phosphate, 0.0-1 wt% of magnesium sulfate and the balance of deionized water are uniformly mixed, 1-3mol/L of sodium carbonate is adopted to adjust the pH of the solution to be neutral, and the solution is sterilized for 10-30min at the temperature of 110-;

s3, fermentation: inoculating the seed liquid A into a fermentation medium A according to the inoculation amount of 2-8 wt%, and fermenting at 30-38 ℃ for 20-30h to obtain a fermentation product A; the preparation method of the fermentation medium A comprises the following steps: uniformly mixing 1-5 wt% of pea polypeptide, 1-4 wt% of lactose, 0.5-2 wt% of peptone, 0.1-1 wt% of dipotassium hydrogen phosphate, 0.05-1 wt% of manganese sulfate, 0.01-0.5 wt% of sodium selenite and the balance of deionized water, adjusting the pH of the solution to be neutral by adopting 1-3mol/L of sodium carbonate, and sterilizing at the temperature of 110-;

s4, collecting and emulsifying bacterial sludge: centrifuging the fermentation product A at 4000-7000rpm, discarding the supernatant, collecting bacterial sludge A, adding the bacterial sludge A into the freeze-drying protective agent A according to the mass ratio of 1 (1-4), and emulsifying at 100-300rpm for 15-40min to obtain an emulsion A; the lyoprotectant A is prepared as follows: uniformly mixing 2-8 wt% of glycerol, 10-30 wt% of skim milk powder, 2-6 wt% of ascorbic acid, 0.5-3 wt% of sodium glutamate, 0.5-2 wt% of prebiotics and the balance of deionized water;

s5, freeze-drying and crushing: freezing the emulsion A at-20-60 deg.C for 2-5h, and freezingDrying at vacuum temperature of-40-60 deg.C and vacuum degree of 0.1-0.5mbar for 10-30h, taking out, pulverizing, and sieving with 10-40 mesh sieve to obtain Bacillus natto lyophilized powder. Aiming at the defects of low viable count and difficult high-density fermentation of the common fermented bacillus natto, the lactobacillus johnsonii fermented by adopting the special fermentation medium and the fermentation method thereof has obviously improved thallus density compared with the common fermentation, and the viable count is not less than 2 multiplied by 10¹⁰cfu/g, which is difficult to achieve and overcome by the prior art, the viable count is greatly improved, and the high-density fermentation of the high viable bacillus natto is realized;

meanwhile, the bergamot extract is added as a nutrient of the bacillus natto, and active substances such as linalyl acetate, limonene, linalool, aldehydes, ketones, esters and the like which are main components of the bergamot extract can effectively promote the growth and the reproduction of the bacillus natto, and can inhibit the growth of mixed bacteria and improve the survival rate of the bacillus natto; in addition, the pea polypeptide is added into the fermentation medium, belongs to the active ingredient of the probiotic preparation of the bioactive peptide, and can play a role of a freeze-drying protective agent, so that the damage of freeze drying on the probiotic is prevented, the live bacteria survival rate of the bacillus natto is improved, the production period of the prepared bacillus natto is shortened, the production cost is reduced, and the production efficiency is improved.

Preferably, the preparation method of the bacillus natto freeze-dried powder comprises the following steps:

s3, fermentation: inoculating the seed liquid A into a fermentation medium A according to the inoculation amount of 2-8 wt%, and fermenting at 30-38 ℃ for 20-30h to obtain a fermentation product A; the preparation method of the fermentation medium A comprises the following steps: 1-5 wt% of pea polypeptide, 1-4 wt% of lactose, 0.5-2 wt% of peptone, 0.1-1 wt% of dipotassium hydrogen phosphate, 0.05-1 wt% of manganese sulfate, 0.01-0.5 wt% of sodium selenite, 0.01-0.8 wt% of nutrient and the balance of deionized water are uniformly mixed, 1-3mol/L of sodium carbonate is adopted to adjust the pH of the solution to be neutral, and the solution is sterilized for 10-30min at the temperature of 110-;

s5, freeze-drying and crushing: freezing the emulsion A at-20-60 ℃ for 2-5h, drying at-40-60 ℃ under vacuum degree of 0.1-0.5mbar for 10-30h, taking out, pulverizing, and sieving with 10-40 mesh sieve to obtain Bacillus natto lyophilized powder.

The nutritional agent in the S3 is a bergamot extract, and the preparation method of the bergamot extract comprises the following steps:

t1, cleaning, drying and crushing bergamot, sieving with a 30-70-mesh sieve, taking 50-100 parts by weight of bergamot powder, adding 800-1200 parts by weight of aqueous solution for mixing, adding 0.01-1 wt% of pectinase, adjusting the pH value to 3-5 by using 1-3mol/L hydrochloric acid, performing enzymolysis at constant temperature of 40-50 ℃ for 20-30min, continuously stirring at room temperature and 200-400rpm for 30-60min, and filtering to obtain filter residue A and filtrate A;

t2, taking all filter residues A of T1, adding 800-1200 parts by weight of 30-60 wt% ethanol aqueous solution for mixing, placing the mixture at 40-60 ℃, ultrasonic power of 300-500W and ultrasonic frequency of 30-60kHz for ultrasonic 0.5-2h, filtering to obtain filtrate B, combining the filtrate A and the filtrate B, and reducing the concentration to 1/10-1/20 of the original volume at 80 ℃ to obtain concentrated solution;

t3, vacuum drying the concentrated solution of T2 at 70-90 deg.C to constant weight to obtain fructus Citri Sarcodactylis extract.

The prebiotics in the S4 are any one of isomaltooligosaccharide, fructo-oligosaccharide, malto-oligosaccharide, raffinose and isomerized lactose.

The preparation method of the lactobacillus johnsonii freeze-dried powder comprises the following steps:

w1, strain activation: taking Lactobacillus johnsonii to streak on a solid culture medium B, and statically culturing for 30-48h at the constant temperature of 30-45 ℃ to obtain activated Lactobacillus johnsonii; the solid medium B was prepared as follows: uniformly mixing 1-5 wt% of peptone, 1-4 wt% of glucose, 0.1-1 wt% of magnesium sulfate, 1-5 wt% of agar powder and the balance of deionized water, and sterilizing at the temperature of 110-;

w2, expanding culture: inoculating activated Lactobacillus johnsonii in a liquid culture medium B, wherein the inoculation amount is 1-5 wt%, and culturing in a shaking table at 32-40 ℃ for 18-36h to obtain a seed solution B; the preparation method of the liquid culture medium B comprises the following steps: the beef extract is 10-20 wt%, the peptone is 10-30 wt%, the glucose is 20-40 wt%, the sodium acetate is 2-8 wt%, the citric acid diamine is 0.5-2 wt%, the dipotassium phosphate is 0.1-1 wt%, the Tween-800.1-1 wt%, the magnesium sulfate is 0.1-0.5 wt%, and the balance is deionized water, and the beef extract is uniformly mixed and sterilized at the temperature of 110-;

w3, fermentation: inoculating the seed liquid B into a fermentation medium B according to the inoculation amount of 2-6 wt%, and performing static fermentation at 30-45 ℃ for 8-18h to obtain a fermentation product B; the preparation method of the fermentation medium B comprises the following steps: the beef extract is 10-20 wt%, the peptone is 10-30 wt%, the glucose is 20-40 wt%, the sodium acetate is 2-8 wt%, the citric acid diamine is 0.5-2 wt%, the dipotassium phosphate is 0.1-1 wt%, the Tween-800.1-1 wt%, the magnesium sulfate is 0.1-0.5 wt%, and the balance is deionized water, and the beef extract is uniformly mixed and sterilized at the temperature of 110-;

w4, bacterial sludge collection and emulsification: centrifuging the fermentation product B at 10000-15000rpm for 10-30min, removing the supernatant, collecting bacterial sludge B, adding the bacterial sludge B into the freeze-drying protective agent B according to the mass ratio of 1 (1-4), and emulsifying at 150-200rpm for 10-30min to obtain an emulsion B; the preparation method of the freeze-drying protective agent B comprises the following steps: uniformly mixing 10-30 wt% of skimmed milk powder, 2-8 wt% of trehalose, 3-8 wt% of glycerol, 1-4 wt% of maltodextrin and the balance of deionized water;

w5, lyophilization and pulverization: freezing the emulsion B at-40 to-50 ℃ for 2-8h, drying at-50 to-70 ℃ under vacuum at 0.1 to 0.5mbar for 40-60h, taking out, pulverizing, and sieving with 10-40 mesh sieve to obtain Lactobacillus johnsonii lyophilized powder.

The preparation method of the akkermansia muciniphila freeze-dried powder comprises the following steps:

x1, strain activation: taking Ackermanella muciniphila to streak on a solid culture medium C, and culturing for 20-28h at the constant temperature of 35-40 ℃ to obtain activated Ackermanella muciniphila; the solid medium C was prepared as follows: uniformly mixing 1-5 wt% of casein peptone, 0.5-2 wt% of glucose, 0.5-2 wt% of fucose, 0.1-1 wt% of threonine, 1-5 wt% of agar powder and the balance of deionized water, adjusting the pH to 6.0-7.0 by using 1-3mol/L hydrochloric acid solution, and sterilizing at the temperature of 110-;

x2, expanding culture: inoculating Ackermanella muciniphila in a liquid culture medium C with the inoculation amount of 1-4 wt%, and culturing in a shaker at 35-40 deg.C for 20-28h to obtain a seed solution C; the preparation method of the liquid culture medium C comprises the following steps: uniformly mixing 10-30 wt% of beef extract, 1-5 wt% of casein peptone, 0.5-2 wt% of glucose, 0.5-2 wt% of fucose, 0.1-0.5 wt% of threonine, 0.1-0.5 wt% of magnesium sulfate and the balance of deionized water, adjusting the pH to 6.0-7.0 by using 1-3mol/L hydrochloric acid solution, and sterilizing at the temperature of 110-;

x3, fermentation: inoculating the seed liquid C into a fermentation culture medium C according to the inoculation amount of 2-8 wt%, and performing static fermentation at 35-40 ℃ for 12-24h to obtain a fermentation product C; the preparation method of the fermentation medium C comprises the following steps: uniformly mixing 10-30 wt% of beef extract, 1-5 wt% of casein peptone, 0.5-2 wt% of glucose, 0.5-2 wt% of fucose, 0.1-0.5 wt% of threonine, 0.1-0.5 wt% of magnesium sulfate and the balance of deionized water, adjusting the pH to 6.0-7.0 by using 1-3mol/L hydrochloric acid solution, and sterilizing at the temperature of 110-;

x4, bacterial sludge collection and emulsification: placing the fermentation product B obtained in the step X3 at 6000-10000rpm for centrifugation for 10-30min, removing the supernatant, collecting bacterial sludge C, adding the bacterial sludge C into the freeze-drying protective agent C according to the mass ratio of 1 (1-4), and emulsifying at 100-300rpm for 10-30min to obtain an emulsion C; the preparation method of the freeze-drying protective agent C comprises the following steps: uniformly mixing 10-30 wt% of skimmed milk powder, 2-8 wt% of glycerol, 3-10 wt% of oligomeric maltose and the balance of deionized water;

x5, lyophilization and pulverization: freezing the emulsion C at-40 to-50 ℃ for 2-8h, drying at-50 to-70 ℃ under a vacuum degree of 0.1 to 0.5mbar for 40-60h, taking out of the bin, crushing, and sieving with a 10-40 mesh sieve to obtain the adhesive protein akkermansia freeze-dried powder.

The preparation method of the probiotic goat milk powder comprises the following steps:

adding whey protein powder, peanut oil, walnut oil, plant nutrient powder, trehalose, L-carnitine, choline tartrate, compound vitamins and compound minerals into goat milk, and uniformly mixing to obtain a mixed material I; preheating the mixed material I to 50-70 ℃, homogenizing at the homogenizing pressure of 200-220bar, heating to 90-100 ℃, and preserving the heat for 3-8s for sterilization to obtain a mixed material II; carrying out triple effect concentration on the mixed material II to 40-50% of the original weight to obtain a mixed material III; spray drying the mixture III, and cooling to room temperature to obtain a mixture IV; adding arachidonic acid, docosahexaenoic acid and a probiotic composition into the mixed material IV and uniformly mixing; and packaging, detecting and warehousing to obtain the product.

The first-effect pressure of the triple-effect concentration is 300-320bar, and the first-effect temperature is 70-75 ℃; the secondary effect pressure is 250-260bar, and the secondary effect temperature is 65-68 ℃; the triple effect temperature is 60-63 ℃; wherein the air inlet temperature of the spray drying is 185-190 ℃, the air outlet temperature is 90-95 ℃, and the air flow rate is 0.2-0.3m³/min。

The invention has the beneficial effects that: the probiotic goat milk powder disclosed by the invention is rich and comprehensive in nutrition and good in taste, and can supplement nutrient elements for a human body in time, improve the gastrointestinal tract function of the human body and improve the immunity of the human body. The composite probiotic composition consists of bacillus natto freeze-dried powder, lactobacillus johnsonii freeze-dried powder and akkermansia muciniphila freeze-dried powder, growth and reproduction of beneficial bacteria in intestinal tracts and intestinal health are facilitated, and the three components synergistically play a role in protecting the intestinal tracts, improving the microbial balance of the intestinal tracts, regulating the intestinal tract function and improving the immunity of organisms.

Detailed Description

The above summary of the present invention is described in further detail below with reference to specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples.

Introduction of some raw materials in this application:

goat milk, which is adopted as goat milk, is standardized, subjected to impurity separation and acceptance, pasteurized and homogenized.

w1, strain activation: taking Lactobacillus johnsonii to streak on a solid culture medium B, and statically culturing for 36h at the constant temperature of 38 ℃ to obtain activated Lactobacillus johnsonii; the solid medium B was prepared as follows: uniformly mixing 3 wt% of peptone, 2 wt% of glucose, 0.5 wt% of magnesium sulfate, 2 wt% of agar powder and the balance of deionized water, and sterilizing at 120 ℃ for 20 min;

w2, expanding culture: inoculating activated lactobacillus johnsonii into a liquid culture medium B, wherein the inoculation amount is 2 wt%, and culturing in a shaking table at 36 ℃ for 24 hours to obtain a seed solution B; the preparation method of the liquid culture medium B comprises the following steps: uniformly mixing 15 wt% of beef extract, 15 wt% of peptone, 30 wt% of glucose, 5 wt% of sodium acetate, 1 wt% of diamine citrate, 0.5 wt% of dipotassium phosphate, 0.2 wt% of tween-800.5, 0.2 wt% of magnesium sulfate and the balance of deionized water, and sterilizing at 120 ℃ for 20 min;

w3, fermentation: inoculating the seed liquid B into a fermentation medium B according to the inoculation amount of 4 wt%, and performing static fermentation for 10h at 36 ℃ to obtain a fermentation product B; the preparation method of the fermentation medium B comprises the following steps: 15 wt% of beef extract, 15 wt% of peptone, 30 wt% of glucose, 5 wt% of sodium acetate, 1 wt% of diamine citrate, 0.5 wt% of dipotassium phosphate, 0.2 wt% of tween-800.5 and 0.2 wt% of magnesium sulfate, and sterilizing at 120 ℃ for 20 min;

w4, bacterial sludge collection and emulsification: centrifuging the fermentation product B at 12000rpm for 15min, removing supernatant, collecting bacterial sludge B, adding the bacterial sludge B into the freeze-drying protective agent B according to the mass ratio of 1:2, and emulsifying at 180rpm for 15min to obtain an emulsion B; the preparation method of the freeze-drying protective agent B comprises the following steps: uniformly mixing 20 wt% of skimmed milk powder, 5 wt% of trehalose, 5 wt% of glycerol, 2 wt% of maltodextrin and the balance of deionized water;

w5, lyophilization and pulverization: freezing the emulsion B at-45 deg.C for 3h, drying at-60 deg.C under vacuum degree of 0.2mbar for 48h, taking out, pulverizing, and sieving with 20 mesh sieve to obtain Lactobacillus johnsonii lyophilized powder.

Lactobacillus johnsonii, deposited in China center for type culture Collection with a collection number of CCTCC NO: m20191013.

x1, strain activation: taking Ackermanella muciniphila to streak on a solid culture medium C, and culturing for 24h at the constant temperature of 37 ℃ to obtain activated Ackermanella muciniphila; the solid medium C was prepared as follows: uniformly mixing 3 wt% of casein peptone, 1 wt% of glucose, 1 wt% of fucose, 0.2 wt% of threonine, 3 wt% of agar powder and the balance of deionized water, adjusting the pH to 6.5 by using 2mol/L hydrochloric acid, and sterilizing at 120 ℃ for 20 min;

x2, expanding culture: inoculating the activated akkermansia muciniphila in a liquid culture medium C, wherein the inoculation amount is 2 wt%, and culturing in a shaker at 37 ℃ for 24h to obtain a seed solution C; the preparation method of the liquid culture medium C comprises the following steps: uniformly mixing 15 wt% of beef extract, 3 wt% of casein peptone, 1 wt% of glucose, 1 wt% of fucose, 0.2 wt% of threonine, 0.2 wt% of magnesium sulfate and the balance of deionized water, adjusting the pH to 6.5 by using 2mol/L hydrochloric acid, and sterilizing at 120 ℃ for 20 min;

x3, fermentation: inoculating the seed solution C into a fermentation medium C according to the inoculation amount of 4 wt%, and performing static fermentation at 37 ℃ for 18h to obtain a fermentation product C; the preparation method of the fermentation medium C comprises the following steps: uniformly mixing 15 wt% of beef extract, 3 wt% of casein peptone, 1 wt% of glucose, 1 wt% of fucose, 0.2 wt% of threonine, 0.2 wt% of magnesium sulfate and the balance of deionized water, adjusting the pH to 6.5 by using 2mol/L hydrochloric acid, and sterilizing at 120 ℃ for 20 min;

x4, bacterial sludge collection and emulsification: placing the fermentation product B obtained in the step X3 at 8000rpm, centrifuging for 20min, removing supernatant, collecting bacterial sludge C, adding the bacterial sludge C into the freeze-drying protective agent C according to the mass ratio of 1:2, and emulsifying at 200rpm for 15min to obtain emulsion C; the preparation method of the freeze-drying protective agent C comprises the following steps: uniformly mixing 20 wt% of skimmed milk powder, 5 wt% of glycerol, 5 wt% of oligomeric maltose and the balance of deionized water;

x5, lyophilization and pulverization: freezing the emulsion C at-45 deg.C for 3h, drying at-60 deg.C under vacuum degree of 0.2mbar for 48h, taking out, pulverizing, and sieving with 20 mesh sieve to obtain the lyophilized powder of Acermanniella muciniphila.

Akkermansia muciniphila, purchased from the china industrial microbial strain collection management center, under the strain collection number: CICC 24917.

Bacillus natto, Bacillus subtilis, is preserved in China general microbiological culture Collection center, and the preservation number is: CGMCC No. 5769.

Defatted milk powder and whey protein powder are purchased from Zhengzhou Bai Si Te food additive Co., Ltd, and are in food grade.

Organic inulin is purchased from food science and technology limited, mingjie, henna, in an amount of: 99 percent and is in food grade.

The pea polypeptide is purchased from Gansu Probiotics and auspicious biotechnology limited, the total protein is more than or equal to 90 percent, the peptide content is more than or equal to 80 percent, the molecular weight is less than 1000, and the pea polypeptide is in food grade.

Bergamot, burdock powder, cassia seed powder and tuckahoe powder are purchased in the Bozhou city and are measured by the cooperative society of the Xianggang Chinese medicinal plant planting specialty in urban areas.

Fucose and trehalose are purchased from Zhengzhou Bai Si Te food additive Co Ltd, and are in food grade.

Pectinase was purchased from huizhou yuan biotechnology ltd, enzyme activity: 10 ten thousand U/g and food grade.

Isomaltooligosaccharides were purchased from seian university harvest biotechnology limited, type: IMO-500, Cat number: 45000, content of active substance: 99 percent and is in food grade.

Example 1

The probiotic goat milk powder is prepared from the following raw materials: 400 parts of goat milk, 60 parts of whey protein powder, 3 parts of peanut oil, 2 parts of walnut oil, 5 parts of plant nutrient powder, 0.5 part of trehalose, 0.06 part of L-carnitine, 0.05 part of choline tartrate, 0.2 part of vitamin complex, 0.2 part of compound mineral, 0.3 part of arachidonic acid, 0.2 part of docosahexaenoic acid and 2 parts of probiotic composition.

adding whey protein powder, peanut oil, walnut oil, plant nutrient powder, trehalose, L-carnitine, choline tartrate, compound vitamins and compound mineral substances into goat milk according to a formula, and uniformly mixing to obtain a mixed material I; preheating the mixed material I to 60 ℃, homogenizing under the homogenizing pressure of 210bar, heating to 95 ℃, preserving heat for 5s, and sterilizing to obtain a mixed material II; carrying out triple effect concentration on the mixed material II to 48% of the original weight to obtain a mixed material III; wherein the first effect pressure of the triple effect concentration is 310bar, and the first effect temperature is 72 ℃; the double-effect pressure is 250bar, and the double-effect temperature is 66 ℃; the triple effect temperature is 62 ℃; spray drying the mixture III, and cooling to room temperature to obtain a mixture IV; wherein the air inlet temperature of spray drying is 188 ℃, the air outlet temperature is 92 ℃, and the air flow rate is 0.23m³Min; adding arachidonic acid, docosahexaenoic acid and a probiotic composition into the mixed material IV and uniformly mixing; and packaging, detecting and warehousing to obtain the product.

The compound vitamin is a mixture of L-ascorbic acid, retinol, cholecalciferol, folic acid and D-calcium pantothenate according to a mass ratio of 5:2:1:0.5: 0.5; the compound mineral is a mixture of calcium chloride, ferrous sulfate, magnesium sulfate and sodium selenate according to a mass ratio of 15:5:3: 0.06; the plant nutrition powder is a mixture of burdock powder, cassia seed powder and tuckahoe powder according to a mass ratio of 1:1: 3.

The probiotic composition is bacillus natto freeze-dried powder; the preparation method of the bacillus natto freeze-dried powder comprises the following steps:

s1, strain activation: marking out the bacillus natto on a solid culture medium A, and culturing for 24 hours at 37 ℃ to obtain activated bacillus natto; the preparation method of the solid culture medium A comprises the following steps: uniformly mixing 3 wt% of peptone, 2 wt% of glucose, 0.5 wt% of NaCl, 2 wt% of agar powder and the balance of deionized water, and sterilizing at 120 ℃ for 20 min;

s2, expanding culture: inoculating activated bacillus natto into a liquid culture medium A, wherein the inoculation amount is 2 wt%, and performing shaking culture at 38 ℃ and 300rpm for 12 hours to obtain a seed solution A; the preparation method of the liquid culture medium A comprises the following steps: uniformly mixing 10 wt% of skim milk powder, 2 wt% of organic inulin, 1 wt% of potassium dihydrogen phosphate, 0.2 wt% of magnesium sulfate and the balance of deionized water, adjusting the pH of the solution to be neutral by adopting 2mol/L sodium carbonate, and sterilizing at 120 ℃ for 20 min;

s3, fermentation: inoculating the seed liquid A into a fermentation medium A according to the inoculation amount of 5 wt%, and fermenting at 35 ℃ for 24h to obtain a fermentation product A; the preparation method of the fermentation medium A comprises the following steps: uniformly mixing 2 wt% of pea polypeptide, 2 wt% of lactose, 1 wt% of peptone, 0.2 wt% of dipotassium hydrogen phosphate, 0.1 wt% of manganese sulfate, 0.05 wt% of sodium selenite, 0.1 wt% of a nutrient and the balance of deionized water, adjusting the pH of the solution to be neutral by adopting 2mol/L sodium carbonate, and sterilizing at 120 ℃ for 20 min;

s4, collecting and emulsifying bacterial sludge: centrifuging the fermentation product A at 5000rpm for 15min, removing supernatant, collecting bacterial sludge A, adding the bacterial sludge A into the freeze-drying protective agent A according to the mass ratio of 1:2, and emulsifying at 200rpm for 30min to obtain an emulsion A; the lyoprotectant A is prepared as follows: uniformly mixing 5 wt% of glycerol, 20 wt% of skim milk powder, 4 wt% of ascorbic acid, 1.5 wt% of sodium glutamate, 1 wt% of isomaltooligosaccharide and the balance of deionized water;

s5, freeze-drying and crushing: freezing the emulsion A at-40 deg.C for 3h, drying at-50 deg.C under vacuum degree of 0.2mbar for 20h, taking out, pulverizing, and sieving with 20 mesh sieve to obtain Bacillus natto lyophilized powder.

The nutritional agent is a bergamot extract, and the preparation method of the bergamot extract comprises the following steps:

t1, cleaning, drying and crushing bergamot, sieving with a 50-mesh sieve, taking 80 parts by weight of bergamot powder, adding 1000 parts by weight of deionized water solution for mixing, adding 0.1 wt% of pectinase, adjusting pH to 4 by using 2mol/L hydrochloric acid, carrying out enzymolysis at a constant temperature of 45 ℃ for 25min, continuously stirring at room temperature and 300rpm for 40min, and filtering to obtain filter residue A and filtrate A;

t2, taking all filter residues A of T1, adding 1000 parts by weight of 40 wt% ethanol aqueous solution, mixing, putting into ultrasonic waves with the ultrasonic power of 400W and the ultrasonic frequency of 45kHz at 50 ℃ for 1h, filtering to obtain filtrate B, combining the filtrate A and the filtrate B, and reducing and concentrating the filtrate A and the filtrate B to 1/15 of the original volume at 80 ℃ to obtain concentrated solution;

t3, vacuum drying the concentrated solution of T2 at 80 deg.C to constant weight to obtain fructus Citri Sarcodactylis extract.

Example 2

Essentially the same as example 1, except that: the probiotic composition consists of 50 wt% of bacillus natto freeze-dried powder and 50 wt% of lactobacillus johnsonii freeze-dried powder. The preparation method of the bacillus natto freeze-dried powder is the same as that of the example 1.

Example 3

Essentially the same as example 1, except that: the probiotic composition consists of 50 wt% of bacillus natto freeze-dried powder and 50 wt% of akmansia muciniphila freeze-dried powder. The preparation method of the bacillus natto freeze-dried powder is the same as that of the example 1.

Example 4

Essentially the same as example 1, except that: the probiotic composition consists of 50 wt% of bacillus natto freeze-dried powder, 25 wt% of lactobacillus johnsonii freeze-dried powder and 25 wt% of akkermansia muciniphila freeze-dried powder. The preparation method of the bacillus natto freeze-dried powder is the same as that of the example 1. Wherein the viable count of Bacillus natto lyophilized powder is 3.7 × 10¹¹cfu/g, viable count in Lactobacillus johnsonii freeze-dried powder 8.3 x 10¹¹cfu/g, viable count in Ackermanella mucronata freeze-dried powder 2.0 x 10¹¹cfu/g。

Example 5

Essentially the same as example 1, except that: the preparation method of the bacillus natto freeze-dried powder comprises the following steps:

s3, fermentation: inoculating the seed liquid A into a fermentation medium A according to the inoculation amount of 5 wt%, and fermenting at 35 ℃ for 24h to obtain a fermentation product A; the preparation method of the fermentation medium A comprises the following steps: uniformly mixing 2 wt% of pea polypeptide, 2 wt% of lactose, 1 wt% of peptone, 0.2 wt% of dipotassium hydrogen phosphate, 0.1 wt% of manganese sulfate, 0.05 wt% of sodium selenite and the balance of deionized water, adjusting the pH of the solution to be neutral by adopting 2mol/L sodium carbonate, and sterilizing at 120 ℃ for 20 min;

Example 6

s3, fermentation: inoculating the seed liquid A into a fermentation medium A according to the inoculation amount of 5 wt%, and fermenting at 35 ℃ for 24h to obtain a fermentation product A; the preparation method of the fermentation medium A comprises the following steps: uniformly mixing 2 wt% of lactose, 1 wt% of peptone, 0.2 wt% of dipotassium phosphate, 0.1 wt% of manganese sulfate, 0.05 wt% of sodium selenite, 0.1 wt% of nutrient and the balance of deionized water, adjusting the pH of the solution to be neutral by adopting 2mol/L sodium carbonate, and sterilizing at 120 ℃ for 20 min;

Comparative example 1

Essentially the same as example 4, except that: the probiotic goat milk powder is prepared from the following raw materials: 400 parts of goat milk, 60 parts of whey protein powder, 3 parts of peanut oil, 2 parts of walnut oil, 0.5 part of trehalose, 0.06 part of L-carnitine, 0.05 part of choline tartrate, 0.2 part of vitamin complex, 0.2 part of compound mineral, 0.3 part of arachidonic acid, 0.2 part of docosahexaenoic acid and 2 parts of probiotic composition.

Comparative example 2

Essentially the same as example 4, except that: the probiotic goat milk powder is prepared from the following raw materials: 400 parts of goat milk, 60 parts of whey protein powder, 5 parts of plant nutrient powder, 0.5 part of trehalose, 0.06 part of L-carnitine, 0.05 part of choline tartrate, 0.2 part of compound vitamin, 0.2 part of compound mineral, 0.3 part of arachidonic acid, 0.2 part of docosahexaenoic acid and 2 parts of probiotic composition.

Test example 1

And (3) evaluating the functions of the probiotic goat milk powder: 60 SPF-grade Wistar rats of 14d birth were selected and randomly divided into 3 groups of 10 males and females in each group. The probiotics goat milk powder (prepared by warm boiled water at 37 ℃ to be 1g/100mL) of the example 4 and the comparative examples 1-2 is respectively fed, and the goat milk powder is freely drunk for 7 days. After the test, the number of stools in the mouse 1d was observed, and the change in body weight before and after the test was counted.

TABLE 1 functional evaluation results of probiotic goat milk powder

	Weight gain g	Stool frequency, times/d
			Example 4	10.2	1.8
Comparative example 1	8.8	1.3
			Comparative example 2	9.5	1.5

Test example 2

And (3) measuring the viable count before freeze-drying: the Bacillus natto bacterial sludge, the Lactobacillus johnsonii bacterial sludge and the Ackermansia muciniphila bacterial sludge collected in the above examples 1 and 5-6 are taken and put into 10ml of 0.85% (m/v) physiological saline to be shaken evenly, and the viable count is measured by a gradient dilution method and an agar culture medium.

Fully shaking the cultured bacteria liquid on a clean bench, and continuously diluting all samples by 10 times with the dilution degree of 10^-1、10^-2、10^-3、10^-4、10^-5、10^-6And 10^-7. Selecting appropriate dilution, uniformly inoculating 1mL of the dilution into a modified MRS agar culture medium which is sterilized, cooled to about 45 ℃, shaking uniformly, culturing at 37 ℃ for 48h, selecting the dilution with about 100 colony counts, selecting 2 dilutions for each sample, performing 3 repeated averaging on each dilution, and calculating the unit viable count (cfu/g).

And (3) measuring the viable count after freeze-drying: taking 1g of Bacillus natto freeze-dried powder, Lactobacillus johnsonii freeze-dried powder and Ackermannheimia muciniphila freeze-dried powder, adding 10mL of 0.85% (m/v) physiological saline into a centrifugal tube for restoration, fully oscillating until the freeze-dried powder is completely dissolved, and respectively diluting with 10 degrees of dilution^-1、10^-2、10^-3、10^-4、10^-5、10^-6And 10^-7. Selecting appropriate dilution, uniformly inoculating 1mL of the diluted solution into a modified MRS agar culture medium which is sterilized and then cooled to about 45 ℃, shaking uniformly, and culturing at 37 ℃ for 48h, wherein the dilution with about 100 colonies is selected.

TABLE 2 detection results of viable count before lyophilization

	Bacillus natto (cfu/g)
		Example 1	9.8×10¹¹
Example 5	4.1×10¹⁰
		Example 6	6.8×10¹⁰

TABLE 3 detection results of viable count after lyophilization

	Bacillus natto (cfu/g)
		Example 1	3.7×10¹¹
Example 5	1.4×10¹⁰
		Example 6	2.8×10¹⁰

From the results, the content of the viable count can be effectively improved by adding the bergamot extract serving as a nutrient in the preparation process of the bacillus natto, the content of the viable count can also be improved to a certain extent by adding the pea polypeptide, and the viable count is also effectively improved after freeze-drying. The reason is that the main components of the bergamot extract, such as linalyl acetate, limonene, linalool, aldehydes, ketones, esters and other active substances, can effectively promote the growth and the propagation of the bacillus natto, and can inhibit the growth of mixed bacteria and improve the survival rate of the bacillus natto; in addition, the pea polypeptide belongs to the effective component of the probiotic preparation of the bioactive peptide, and simultaneously can play the role of a freeze-drying protective agent, so that the damage of freeze-drying to probiotics is prevented, the survival rate of viable bacteria of bacillus natto is improved, the defatted milk powder in the freeze-dried powder can prevent the cell damage by stabilizing the composition of cell membranes, the defatted milk powder also contains protein and can provide a protective layer for the cells, in addition, a porous structure is generated in the freeze-drying process, so that the dehydration is easier, the survival rate of thalli can be improved, the excessive moisture of the product is removed by freeze-drying, and the stability of the finished product is improved; meanwhile, the pea polypeptide and the probiotics are freeze-dried together, so that the activity of the probiotics is kept, the effective viable count of the finished product is higher, and the activity of the pea polypeptide can be improved. Meanwhile, the probiotic composition prepared by the invention has a production period, so that the production cost is reduced and the production efficiency is improved.

Test example 3

Rat testing: SPF-class Wistar rats weighing 150 + -20 g were selected and randomly divided into experimental groups (examples 1-6) of 10 males and females per group. The rats are numbered before the test, the probiotic composition in the example 1-6 is prepared into a probiotic composition solution with the concentration of 100mg/L by warm boiled water at 37 ℃, the probiotic composition solution in the example 1-6 is perfused for 3 times a day by an experimental group of rats with the dosage of 0.5mL each time, warm boiled water at 37 ℃ is perfused for 3 times a day by a blank group of rats, the rats in the experimental group and the blank group freely eat and drink water and are continuously fed for 30 days, and the performance, behavior, toxicity performance and death condition of the rats are observed every day.

1. After feeding for 30 days, fasting was performed for 10 hours, and blood was collected from abdominal aorta of rats, serum was separated, and Total Cholesterol (TCH), Triglyceride (TG) and blood sugar (GLU) contents of rats were measured using an enzymatic kit.

TABLE 4 measurement of Total Cholesterol (TCH), Triglyceride (TG) and blood sugar (GLU)

	Total cholesterol (mmol-L)	Triglyceride (mmol/L)	Blood sugar (mmol/L)
				Example 1	1.93	0.49	5.26
Example 2	1.66	0.41	4.92
				Example 3	1.64	0.39	4.93
Example 4	1.43	0.31	4.37
				Example 5	2.19	0.53	5.49
Example 6	2.23	0.54	5.53
				Blank group	3.72	0.68	6.41

2. Mouse peripheral blood immune cells and peritoneal macrophages: separately collecting eyeballs of mice in blank group and test group, collecting total peripheral blood (with mouse hemorrhagic shock as standard), and collecting with Tris-NH₄After the CI solution hypotonically destroys the red blood cells, immune cells are counted, and then the mice after blood collection are dissected on a clean workbench, and abdominal cavity macrophages are taken for counting.

TABLE 5 blood immune cell and peritoneal macrophage assay results

From the results, the probiotic composition prepared by the invention has good effects of reducing blood sugar and blood fat, and also has good immune effect.

Claims

1. The probiotic goat milk powder is characterized by comprising the following raw materials: goat milk, whey protein powder, peanut oil, walnut oil, plant nutrition powder, trehalose, L-carnitine, choline tartrate, compound vitamins, compound minerals, arachidonic acid, docosahexaenoic acid and a probiotic composition.

2. The probiotic goat milk powder as claimed in claim 1, which is characterized by comprising the following raw materials in parts by weight: 400 parts of goat milk, 30-60 parts of whey protein powder, 1-3 parts of peanut oil, 1-2 parts of walnut oil, 3-5 parts of plant nutrient powder, 0.3-0.5 part of trehalose, 0.05-0.08 part of L-carnitine, 0.03-0.05 part of choline tartrate, 0.1-0.2 part of vitamin complex, 0.1-0.2 part of compound mineral, 0.1-0.3 part of arachidonic acid, 0.1-0.2 part of docosahexaenoic acid and 1-2 parts of probiotic composition.

3. The probiotic goat milk powder as claimed in claim 1 or 2, wherein the vitamin complex is a mixture of L-ascorbic acid, retinol, cholecalciferol, folic acid, and D-calcium pantothenate in a mass ratio of 5 (1-2): 0.5-1): 0.3-0.5.

4. The probiotic goat milk powder as claimed in claim 1 or 2, wherein the compound mineral is a mixture of calcium chloride, ferrous sulfate, magnesium sulfate and sodium selenate in a mass ratio of 15 (1-5) to (1-3) to (0.05-0.1).

5. The probiotic goat milk powder as claimed in claim 1 or 2, wherein the plant nutrition powder is a mixture of burdock powder, cassia seed powder and tuckahoe powder in a mass ratio of (1-2): 3.

6. The probiotic goat milk powder of claim 1 or 2, wherein the probiotic composition consists of 20-60 wt% of bacillus natto lyophilized powder, 20-40 wt% of lactobacillus johnsonii lyophilized powder, and 20-40 wt% of akkermansia muciniphila lyophilized powder.

7. The probiotic goat milk powder of claim 6, wherein the Lactobacillus johnsonii lyophilized powder is prepared by the following method:

8. The probiotic goat milk powder of claim 6, wherein the preparation method of the akkermansia muciniphila freeze-dried powder is as follows:

9. The method for preparing probiotic goat milk powder as claimed in any one of claims 1 to 8, comprising the steps of: adding whey protein powder, peanut oil, walnut oil, plant nutrient powder, trehalose, L-carnitine, choline tartrate, compound vitamins and compound minerals into goat milk, and uniformly mixing to obtain a mixed material I; preheating the mixed material I to 50-70 ℃, homogenizing at the homogenizing pressure of 200-220bar, heating to 90-100 ℃, and preserving the heat for 3-8s for sterilization to obtain a mixed material II; carrying out triple effect concentration on the mixed material II to 40-50% of the original weight to obtain a mixed material III; spray drying the mixture III, and cooling to room temperature to obtain a mixture IV; adding arachidonic acid, docosahexaenoic acid and a probiotic composition into the mixed material IV and uniformly mixing; and packaging, detecting and warehousing to obtain the product.