CN1843385A - Enteral microecological formulation and its preparation process - Google Patents
Enteral microecological formulation and its preparation process Download PDFInfo
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Abstract
The invention provides an intestinal tract micro-ecological preparation and its preparing process, wherein each 100g of the product contains bifidobacteria 1.0-20.0g, Lactobacillus acidophilus 1.0-25.0g, Streptococcus thermophilus 1.0-25.0g, Bulgarian subspecies of Lactobacillus fermentum 1.0-25.0g, saccharomycete 1.0-35.0g, and right amount of isomaltose hypgather, fructooligosaccharide, and many kinds of vitamins.
Description
(1) technical field
What the present invention relates to is a kind of microorganism formulation, specifically a kind of human intestine's microbial ecological agent.The invention still further relates to the production method of this microorganism formulation.
(2) background technology
Probiotic bacteria (Probiotics) speech derives from Latin, looks like for " helping life (for life)." in the sixties in 20th century since antibiotic be extensive use of and animal husbandry in antibiotic use the side effect that causes, people begin to notice the effect of some active bacteria formulations in improving animal health.When Lilly and Stillwell create this speech at first, be to be used to refer to the excretory material that is used to stimulate another kind of protozoacide (Protozoan) growth of a kind of protozoacide.Can improve the animal feed preparation of animal health by promoting the microbial population of animal intestinal tract balance thereby Parker in 1974 then describes those with it, comprise various biological or chemical materials, as antibiotic etc.1992, people such as Havenaar were defined as Probiotics: " active bacteria formulation of forming by single or multiple microorganism, after being applied to the human or animal, can be by improving the composition of host indigenous flora, thereby promote human or animal's health ".This definition has developed this notion of probiotic bacteria from many aspects.At first, it no longer only is confined to intestinal microbial population with the effect of probiotic bacteria, and the flora that also might act on other positions is formed, for example positions such as respiratory tract, urogenital tract and skin; Secondly, it has proposed the notion that probiotic bacteria is used first clearly in human body; The 3rd, probiotic bacteria can be made up of more than one microorganism.Afterwards, in order to distinguish common yoghurt bacterium (or sour milk products) and probiotic bacteria and products thereof, Sehrezenmier etc. (2001) revise the Probiotics definition that Huis in ' t Veld and Havenaar (1992) are proposed, revised " probiotic bacteria " is defined as: contain the sufficient amount viable bacteria, form clear and definite microorganism formulation or product, form the host health effect that helps that produces thereby can change a certain position of host flora by the effect of growing surely.The weak point of this definition is still not distinguish the specific microorganism and the concrete form of product.Definition below present the employing: thus help the single of host health effect or form clear and definite mixed microorganism by the composition generation of growing a certain position of effect change host flora surely.
Probiotics (beneficial natural disposition or prebiotic effect) then be meant certain microorganism formulation or fermented product by regulate host's mucosa and systemic immune function and improve enteral nutrition and colony balance to useful physiological action that the host produced.Although part lactobacillus and bacillus bifidus can not arrive intestinal with the viable bacteria state, and grow surely there, its cell wall components still can be brought into play anti-tumor and become and immunoregulation effect, and for example heat-inactivated Lb.casei LC9018 still has facilitation to health.Metchmikoff was according to his result of study to bulgaricus bacillus (Bulgarianbacillus) in 1907, think that often edible fermentation milk helps the health of human body, Lactobaillus delbruekii subsp.bulgaricus used in this bacterium and the growth of modern yoghourt is very close.Up to now, lactobacillus remains most widely used probiotic bacteria.The lactobacillus that is commonly used for probiotic bacteria at present has L.delbruekiisubsp.bulgaricus, Lb.acidophilus, Lb.casei, Lb.fermentum, Lb.reuteri, Lb.rhamnosus, Lb.plantarum, Lb.lactis, Lb.gasseri, Lb.johnsonii etc.With streptococcus with the form of yoghourt as probiotic bacteria, then start from Fuller (1993), as the S.thermophilus of one of ferment agent for sour milk strain.It also is one of beneficial natural disposition lactobacillus of using always.Lactobacillus with prebiotic effect all is intestinal bacteria usually, and they are considered to have the function of improving human intestines and stomach's digestive tract microecological environment.The scope of prebiotic effect can be divided into three aspects such as nutrition, physiological action and antibacterial action.Research and observed effect in these areas directly caused various food and the application of feedstuff in humans and animals that contain lactobacillus.Lactobacillus also has potential immune assosting effect, and oral lactobacillus can excite mucosa and systemic immunoreation.
Bacillus bifidus and bacillus acidophilus are inherent probioticss in the human body, and bacillus bifidus and bacillus acidophilus account for more than 95% of probiotic bacteria in the human body intestinal canal, and they mainly are to play the effect that promotes health by the microecological balance of safeguarding intestinal.The two effect in intestinal is similar, but the position difference that exists, the bacillus acidophilus is at the first half of intestinal, and bacillus bifidus is in the Lower Half of intestinal.Therefore, bacillus bifidus and bacillus acidophilus complement each other, and are safeguarding the health of whole intestinal jointly.
Probiotic bacteria integrates the effect with following several respects:
(1) improve immunity: bacillus bifidus and bacillus acidophilus can produce a large amount of immunoglobulins.Thereby activate the immunocyte of body, make external bacterium be difficult to retain in the body, reach immunoregulation effect.
(2) improve gastrointestinal function: bacillus bifidus and bacillus acidophilus form the protection barrier on the intestinal mucosa surface; resist the invasion and attack destruction of harmful bacterium to intestinal mucosa; can produce acidic materials such as lactic acid and acetic acid; cause the environment that is unfavorable for harmful bacteria growing; suppress growth, the breeding of harmful bacterium, play the effect of regulating intestinal microbial population.
(3) Nutrition: bacillus bifidus and bacillus acidophilus can produce lactic acid and acetic acid after fermenting in human body intestinal canal, can improve the utilization rate of calcium, phosphorus, ferrum, promote the absorption of ferrum and vitamin D, produce vitamin K and vitamin B, produce Lactose enzyme facilitating digestion lactose.
(4) logical intestinal profit just: bacillus bifidus and bacillus acidophilus can produce a large amount of acidic materials such as short-chain fatty acid at enteral, can stimulate intestinal peristalsis promoting, increase feces humidity, play the effect of relieving constipation.
(5) recover because of the normal microorganism species of ruined intestinal behind the use antibiotic: antibiotic is the key factor that causes alteration of intestinal flora.Antibiotic has also been killed a lot of probioticss in the kill harmful bacterium, in time replenish bifidus bacillus and bacillus acidophilus, is very important to helping intestinal recovery colony balance, raising alimentation ability.
(6) cholesterol reducing, prevent arteriosclerosis.
(7) suppress endotoxic generation, slow down aging.
(8) generation of cancerigenic factor in the inhibition intestinal, the generation of prophylaxis of tumours.
(9) radioprotective etc.
Therefore, the research of enteral microecological formulation has become the research emphasis in our times microbiology field, and the research and development meeting of probiotic products brings positive promotion meaning to human beings'health, therefore has important economic value and social benefit.
(3) summary of the invention
The object of the present invention is to provide a kind of compound micro-ecological preparation, the present invention also aims to provide the compound method of this compound micro-ecological preparation.
The object of the present invention is achieved like this: percentage ratio related among the present invention all is weight percentage.
Contain bifidobacterium bifidum 1.0g-20.0, bacillus acidophilus 1.0-25.0g, streptococcus thermophilus 1.0-25.0g, lactobacillus delbruockii subspecies bulgaricus 1.0-25.0g, yeast 1.0-35.0g and oligomeric isomaltose, oligofructose, multivitamin in every 100g product.
Can also also have Bacillus subtillis 1.0-25.0g in every 100g product.
Can also also have natto bacillus 1.0-25.0g in every 100g product.
Product of the present invention adopts following method to prepare:
1, the cultivation of bifidobacterium bifidum, bacillus acidophilus, streptococcus thermophilus, lactobacillus delbruockii subspecies bulgaricus:
Culture medium prescription: Semen Maydis pulp 0.1-15.0 gram, soybean peptide 0.1-15.0 gram, the 1.0%-35% of Tomato juice, whey powder 0.1-15.0 gram, Semen sojae atricolor is soaked juice 1.0%-35%, glucose 0.5-15.0 gram, lactose 0.5-15.0 gram, pancreatin hydrolysis casein 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-20.0 gram, yeast extract 0.1-25.0 gram, sodium citrate 0.01-5.0 gram, sodium acetate 0.01-5.5 gram, dipotassium hydrogen phosphate 0.01-5.5 gram, magnesium sulfate 0.1-2.5 gram, L-cysteine hydrochloride 0.001-2.5 gram, tween 80 0.1-10.0 milliliter, add water and be settled to 1000 milliliters, pH5.0-7.5 sterilized 15 minutes for 105 ℃.
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 10-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 10-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 10-72 hour for 20 ℃-45 ℃.
2, the cultivation of Bacillus subtillis, natto bacillus:
1000 milliliters of culture medium prescription: soybean peptide 0.1-15.0 grams, glucose 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-15.0 gram, peptone 0.1-25 gram, Semen Maydis pulp 0.1-15.0 gram, sodium chloride 0.05-5.0 gram, water, pH5.0-7.5,105 ℃ of sterilizations 15 minutes.
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 16-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 18-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 18-72 hour for 20 ℃-45 ℃.
3, cultivate to finish after, carry out centrifugally with centrifuge 3500-8000r/min, wash 1-2 time with sterilized water, obtain active bacterium mud.
4, the preparation of protection liquid: take by weighing sodium glutamate 0.1-15.0 gram, lactose 0.1 gram-20.0 grams, glycerol 0.1-25.0 gram, polyvinylpyrrolidone 0.1-20.0 gram, Vc 0.01-15.0 gram, water 100mL; heated and stirred is dissolved it; then; 105 ℃ of temperature were sterilized 15 minutes; be cooled to 10 ℃-35 ℃, this solution is protection liquid.
5, the preparation of active bacterium mud-protection liquid: under aseptic technique, take by weighing a certain amount of active bacterium mud and add in the protection liquid active bacterium mud: protection liquid should be 0.5: 1-8: 1, thoroughly stir, and this is an A liquid.
6, microcapsule embedded dose preparation: take by weighing sodium alginate 0.1-15.0 gram, kappa-carrageenan 0.1-10.0 gram, add 100 milliliters in water, stirred 10 minutes, static 2-6 hour, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is an embedding liquid.
7, the preparation of embedding consolidation liquid: take by weighing calcium chloride 0.1-15.0 gram, potassium chloride 0.1-10.0 gram, add 100 milliliters in water, thoroughly dissolving, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is the embedding consolidation liquid.
8, under aseptic technique, take by weighing a certain amount of A liquid and embedding liquid simultaneously, A liquid: embedding liquid is 0.1: 1-15: 1, thoroughly stir, and this is a B liquid.
B liquid is poured in the embedding ejector, and high pressure injects consolidation liquid, and need constantly stir, and the ratio of B liquid and embedding consolidation liquid is by 1: 1-1: 5, and the embedding cured through 2-8 hour is filtered, and washs 1 time with sterilized water.Then, adopt Freeze Drying Technique to carry out lyophilization again, make single Powdered active microorganism mycopowder, pulverize again in case of necessity.
9, the preparation of active microbiological preparations: with the active mycopowder of mentioned microorganism, be mixed in proportion, be equipped with oligomeric isomaltose, multivitamin and other prebioticses again and make active microbiological preparations.
This product can improve the quantity of the probiotic bacteria in the intestinal, improves microecological environment, prevents growing of harmful bacterium, strengthens the resistance against diseases of intestinal, and loosening bowel to relieve constipation guarantees that intestinal is in a kind of health status, promotes human body key health.Quantity and the activity of probiotic bacteria and the preservation time of probiotic bacteria of the more effective assurance probiotics viable bacteria of this preparation method energy.
(4) specific embodiment
For a more detailed description to the present invention for example below:
Prescription 1:(content: g/100g) bifidobacterium bifidum 1.0g, bacillus acidophilus 1.0g, streptococcus thermophilus 2.0g, Lactobacillus bulgaricus 2.5g, yeast 3.5g, oligomeric isomaltose 50.5g, milk powder 38.15g, VB12mg, VB22mg, VB62mg, VC200mg, folic acid 500 μ g, protein sugar 0.01g, strawberry essence 0.04g.
Prescription 2: bifidobacterium bifidum 1.0g, bacillus acidophilus 2.5g, streptococcus thermophilus 2.5g, Lactobacillus bulgaricus 2.5g, Bacillus subtillis 2.0g, yeast 6.5g, oligomeric isomaltose 50.5g, soybean protein powder 10.5g, edible cellulose 10.0g, VB12mg, VB22mg, beta-carotene 2mg, VE2mg, VC500mg, folic acid 500 μ g, xylitol 10.5g, strawberry essence 0.04g.
Prescription 3: natto bacillus 2.5g, bifidobacterium bifidum 1.0g, bacillus acidophilus 2.0g, streptococcus thermophilus 2.0g, Lactobacillus bulgaricus 2.5g, Bacillus subtillis 2.0g, yeast 6.5g, oligomeric isomaltose 30.5g, oligofructose 20.0g, soybean protein powder 10.0g, edible cellulose 10.0g, VB12mg, VB22mg, VE 2mg, VC 500mg, folic acid 500 μ g, xylitol 10.5g, strawberry essence 0.04g.
Prescription 4: bifidobacterium bifidum 1.0g, bacillus acidophilus 2.0g, streptococcus thermophilus 2.0g, Lactobacillus bulgaricus 2.5g, Bacillus subtillis 2.0g, yeast 6.5g, oligomeric isomaltose 20.0g, oligofructose 10.0g, soybean protein powder 40.5g, lysine 1.5g, VB12mg, VB22mg, VB62mg, VC 500mg, folic acid 500 μ g, CaCO35mg, ZnSO42mg, xylitol 10.5g strawberry essence 0.04g.
Prescription 5: natto bacillus 15g, bifidobacterium bifidum 5g, bacillus acidophilus 10g, streptococcus thermophilus 10g, Lactobacillus bulgaricus 15g, Bacillus subtillis 10g, yeast 15g, oligomeric isomaltose 4.5g, oligofructose 2.0g, soybean protein powder 2.0g, edible cellulose 1.0g, VB12mg, VB22mg, VE2mg, VC500mg, folic acid 500 μ g, xylitol 10.5g, strawberry essence 0.04g.
Total viable count is 2.0 * 10 in the product
4Cfu/g-2.0 * 10
10Cfu/g ,-18 ℃ of-30 ℃ of preservations, storage life is 3-18 month.
Product of the present invention is to adopt following method to prepare:
1, the cultivation of bifidobacterium bifidum, bacillus acidophilus, streptococcus thermophilus, lactobacillus delbruockii subspecies bulgaricus:
Culture medium prescription: Semen Maydis pulp 0.1-15.0 gram, soybean peptide 0.1-15.0 gram, the 1.0%-35% of Tomato juice, whey powder 0.1-15.0 gram, Semen sojae atricolor is soaked juice 1.0%-35%, glucose 0.5-15.0 gram, lactose 0.5-15.0 gram, pancreatin hydrolysis casein 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-20.0 gram, yeast extract 0.1-25.0 gram, sodium citrate 0.01-5.0 gram, sodium acetate 0.01-5.5 gram, dipotassium hydrogen phosphate 0.01-5.5 gram, magnesium sulfate 0.1-2.5 gram, L-cysteine hydrochloride 0.001-2.5 gram, tween 80 0.1-10.0 milliliter, add water and be settled to 1000 milliliters, pH5.0-7.5 sterilized 15 minutes for 105 ℃.
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 10-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 10-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 10-72 hour for 20 ℃-45 ℃.
2, the cultivation of Bacillus subtillis, natto bacillus:
1000 milliliters of culture medium prescription: soybean peptide 0.1-15.0 grams, glucose 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-15.0 gram, peptone 0.1-25 gram, Semen Maydis pulp 0.1-15.0 gram, sodium chloride 0.05-5.0 gram, water, pH5.0-7.5,105 ℃ of sterilizations 15 minutes.
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 16-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 18-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 18-72 hour for 20 ℃-45 ℃.
3, cultivate to finish after, carry out centrifugally with centrifuge 3500-8000r/min, wash 1-2 time with sterilized water, obtain active bacterium mud.
4, the preparation of protection liquid: take by weighing sodium glutamate 0.1-15.0 gram, lactose 0.1 gram-20.0 grams, glycerol 0.1-25.0 gram, polyvinylpyrrolidone 0.1-20.0 gram, Vc 0.01-15.0 gram, water 100mL; heated and stirred is dissolved it; then; 105 ℃ of temperature were sterilized 15 minutes; be cooled to 10 ℃-35 ℃, this solution is protection liquid.
5, the preparation of active bacterium mud-protection liquid: under aseptic technique, take by weighing a certain amount of active bacterium mud and add in the protection liquid active bacterium mud: protection liquid should be 0.5: 1-8: 1, thoroughly stir, and this is an A liquid.
6, microcapsule embedded dose preparation: take by weighing sodium alginate 0.1-15.0 gram, kappa-carrageenan 0.1-10.0 gram, add 100 milliliters in water, stirred 10 minutes, static 2-6 hour, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is an embedding liquid.
7, the preparation of embedding consolidation liquid: take by weighing calcium chloride 0.1-15.0 gram, potassium chloride 0.1-10.0 gram, add 100 milliliters in water, thoroughly dissolving, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is the embedding consolidation liquid.
8, under aseptic technique, take by weighing a certain amount of A liquid and embedding liquid simultaneously, A liquid: embedding liquid is 0.1: 1-15: 1, thoroughly stir, and this is a B liquid.
B liquid is poured in the embedding ejector, and high pressure injects consolidation liquid, and need constantly stir, and the ratio of B liquid and embedding consolidation liquid is by 1: 1-1: 5, and the embedding cured through 2-8 hour is filtered, and washs 1 time with sterilized water.Then, adopt Freeze Drying Technique to carry out lyophilization again, make single Powdered active microorganism mycopowder, pulverize again in case of necessity.
9, the preparation of active microbiological preparations: with the active mycopowder of mentioned microorganism, any one mixed by in above-mentioned 4 prescriptions is equipped with oligomeric isomaltose, oligofructose, multivitamin and other prebioticses again and makes active microbiological preparations.
The concrete composition of culture medium can be selected:
1, the cultivation of bifidobacterium bifidum, bacillus acidophilus, streptococcus thermophilus, lactobacillus delbruockii subspecies bulgaricus:
Culture medium prescription: Semen Maydis pulp 3.0 grams, soybean peptide 0.5 gram, Tomato juice 15%, whey powder 5.0 restrain, Semen sojae atricolor is soaked juice 10%, glucose 3.0 grams, lactose 2.0 grams, pancreatin hydrolysis casein 2.0 grams, Carnis Bovis seu Bubali cream 10.0 grams, yeast extract 5.0 grams, sodium citrate 0.02 gram, sodium acetate 0.1 gram, dipotassium hydrogen phosphate 0.5 gram, magnesium sulfate 0.2 gram, L-cysteine hydrochloride 2.5 grams, 3.0 milliliters of tween 80s, add water is settled to 1000 milliliters, pH6.5 sterilized 15 minutes for 105 ℃.
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 24 hours, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 18 hours for 35 ℃, inoculum concentration is 10%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 48 hours for 35 ℃.
2, the cultivation of Bacillus subtillis, natto bacillus:
Culture medium prescription: 1000 milliliters of soybean peptide 1.0 grams, glucose 5.0 grams, Carnis Bovis seu Bubali cream 10 grams, peptone 5.0 grams, Semen Maydis pulp 5.0 grams, sodium chloride 2.0 grams, water, pH7.0,105 ℃ of sterilizations 15 minutes.
Claims (4)
1, a kind of enteral microecological formulation is characterized in that: contain bifidobacterium bifidum 1.0-20.0g, bacillus acidophilus 1.0-25.0g, streptococcus thermophilus 1.0-25.0g, lactobacillus delbruockii subspecies bulgaricus 1.0-25.0g, yeast 1.0-35.0g and oligomeric isomaltose, oligofructose, multivitamin in every 100g product.
2, enteral microecological formulation according to claim 1 is characterized in that: can also also have Bacillus subtillis 1.0-25.0g in every 100g product.
3, enteral microecological formulation according to claim 1 is characterized in that: can also also have natto bacillus 1.0-25.0g in every 100g product.
4, a kind of preparation method of enteral microecological formulation is characterized in that:
(1) cultivation of bifidobacterium bifidum, bacillus acidophilus, streptococcus thermophilus, lactobacillus delbruockii subspecies bulgaricus:
Culture medium prescription: Semen Maydis pulp 0.1-15.0 gram, soybean peptide 0.1-15.0 gram, the 1.0%-35% of Tomato juice, whey powder 0.1-15.0 gram, Semen sojae atricolor is soaked juice 1.0%-35%, glucose 0.5-15.0 gram, lactose 0.5-15.0 gram, pancreatin hydrolysis casein 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-20.0 gram, yeast extract 0.1-25.0 gram, sodium citrate 0.01-5.0 gram, sodium acetate 0.01-5.5 gram, dipotassium hydrogen phosphate 0.01-5.5 gram, magnesium sulfate 0.1-2.5 gram, L-cysteine hydrochloride 0.001-2.5 gram, tween 80 0.1-10.0 milliliter, add water and be settled to 1000 milliliters, pH5.0-7.5 sterilized 15 minutes for 105 ℃;
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 10-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 10-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 10-72 hour for 20 ℃-45 ℃;
(2) cultivation of Bacillus subtillis, natto bacillus:
1000 milliliters of culture medium prescription: soybean peptide 0.1-15.0 grams, glucose 0.1-15.0 gram, Carnis Bovis seu Bubali cream 0.1-15.0 gram, peptone 0.1-25 gram, Semen Maydis pulp 0.1-15.0 gram, sodium chloride 0.05-5.0 gram, water, pH5.0-7.5,105 ℃ of sterilizations 15 minutes;
At first distinguish the culture test tube slant strains, the agar of adding 2% in the slant strains culture medium, cultivated 16-24 hour, insert then in the triangular flask fluid medium and cultivate respectively, cultivated 18-72 hour for 20 ℃-45 ℃, inoculum concentration is 0.5%-25%, carries out amplification culture more step by step, and then carry out bulk fermentation respectively and cultivate, cultivated 18-72 hour for 20 ℃-45 ℃;
(3) cultivate to finish after, carry out centrifugally with centrifuge 3500-8000r/min, wash 1-2 time with sterilized water, obtain active bacterium mud;
(4) preparation of protection liquid: take by weighing sodium glutamate 0.1-15.0 gram, lactose 0.1 gram-20.0 grams, glycerol 0.1-25.0 gram, polyvinylpyrrolidone 0.1-20.0 gram, Vc 0.01-15.0 gram, water 100mL, heated and stirred is dissolved it, then, 105 ℃ of temperature were sterilized 15 minutes, be cooled to 10 ℃-35 ℃, this solution is protection liquid;
(5) preparation of active bacterium mud-protection liquid: under aseptic technique, take by weighing a certain amount of active bacterium mud and add in the protection liquid active bacterium mud: protection liquid should be 0.5: 1-8: 1, thoroughly stir, and this is an A liquid;
(6) microcapsule embedded dose preparation: take by weighing sodium alginate 0.1-15.0 gram, kappa-carrageenan 0.1-10.0 gram, add 100 milliliters in water, stirred 10 minutes, static 2-6 hour, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is an embedding liquid;
(7) preparation of embedding consolidation liquid: take by weighing calcium chloride 0.1-15.0 gram, potassium chloride 0.1-10.0 gram, add 100 milliliters in water, thoroughly dissolving, 105 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃, and this is the embedding consolidation liquid;
(8) under aseptic technique, take by weighing a certain amount of A liquid and embedding liquid simultaneously, A liquid: embedding liquid is 0.1: 1-15: 1, thoroughly stir, and this is a B liquid;
B liquid is poured in the embedding ejector, high pressure injects consolidation liquid, and need constantly to stir, the ratio of B liquid and embedding consolidation liquid is by 1: 1-1: 5, and the embedding cured through 2-8 hour, filter, and, then, adopt Freeze Drying Technique to carry out lyophilization again with sterilized water washing 1 time, make single Powdered active microorganism mycopowder, pulverize again in case of necessity;
(9) preparation of active microbiological preparations: with the active mycopowder of mentioned microorganism, be mixed in proportion, be equipped with oligomeric isomaltose, oligofructose, multivitamin and other prebioticses again and make active microbiological preparations.
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| CN101328469B (en) * | 2008-07-09 | 2010-10-13 | 扬州大学 | Streptococcus thermophilus grx02 having alcoholic liver damage protection function and use thereof |
| CN101486986B (en) * | 2009-02-12 | 2011-06-08 | 上海谱莱生物技术有限公司 | Preparation of freeze-dried Lactobacillus acidophilus powder |
| CN101691568B (en) * | 2009-10-13 | 2012-02-08 | 哈尔滨美华生物技术股份有限公司 | Method of producing edible immobilized active probiotics |
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| CN101574375B (en) * | 2008-05-06 | 2012-12-19 | 上海创博生态工程有限公司 | Lactobacillus health-care capsule and preparation method thereof |
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| CN1051439C (en) * | 1995-08-08 | 2000-04-19 | 包头市环境保护技术开发公司 | Nutrient health food |
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| CN1177039C (en) * | 2002-06-14 | 2004-11-24 | 中国科学院过程工程研究所 | Bacillus adhaerens and its use in preparing health food having thrombolysis property |
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