CN101691568A - Method of producing edible immobilized active probiotics - Google Patents
Method of producing edible immobilized active probiotics Download PDFInfo
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- CN101691568A CN101691568A CN200910073054A CN200910073054A CN101691568A CN 101691568 A CN101691568 A CN 101691568A CN 200910073054 A CN200910073054 A CN 200910073054A CN 200910073054 A CN200910073054 A CN 200910073054A CN 101691568 A CN101691568 A CN 101691568A
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- 239000006041 probiotic Substances 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 10
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- 239000007788 liquid Substances 0.000 claims description 73
- 241000894006 Bacteria Species 0.000 claims description 17
- 230000000529 probiotic effect Effects 0.000 claims description 14
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 13
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 13
- 239000001110 calcium chloride Substances 0.000 claims description 13
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 13
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 claims description 11
- 239000000661 sodium alginate Substances 0.000 claims description 11
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- 239000001888 Peptone Substances 0.000 claims description 5
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- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 5
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- 238000003756 stirring Methods 0.000 claims description 5
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- HRXKRNGNAMMEHJ-UHFFFAOYSA-K trisodium citrate Chemical compound [Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O HRXKRNGNAMMEHJ-UHFFFAOYSA-K 0.000 claims description 5
- 229940038773 trisodium citrate Drugs 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- 241000186660 Lactobacillus Species 0.000 claims description 4
- 238000009395 breeding Methods 0.000 claims description 4
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- ZNOZWUKQPJXOIG-XSBHQQIPSA-L [(2r,3s,4r,5r,6s)-6-[[(1r,3s,4r,5r,8s)-3,4-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-8-yl]oxy]-4-[[(1r,3r,4r,5r,8s)-8-[(2s,3r,4r,5r,6r)-3,4-dihydroxy-6-(hydroxymethyl)-5-sulfonatooxyoxan-2-yl]oxy-4-hydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-5-hydroxy-2-( Chemical compound O[C@@H]1[C@@H](O)[C@@H](OS([O-])(=O)=O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H]2OC[C@H]1O[C@H](O[C@H]1[C@H]([C@@H](CO)O[C@@H](O[C@@H]3[C@@H]4OC[C@H]3O[C@H](O)[C@@H]4O)[C@@H]1O)OS([O-])(=O)=O)[C@@H]2O ZNOZWUKQPJXOIG-XSBHQQIPSA-L 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
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- 241000193830 Bacillus <bacterium> Species 0.000 description 5
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 5
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- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 4
- 210000000936 intestine Anatomy 0.000 description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
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- 206010028980 Neoplasm Diseases 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
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- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention discloses a method of producing edible immobilized active probiotics, comprising the following steps: adding one kind of or various kinds of mature probiotics strains to immobilized carrier solution which is suitable for the growth and the propagation of the probiotics strains according to a certain proportion; adding dropwise the immobilized carrier solution which comprises one kind of or various kinds of probiotics strains into immobilized solution, or adding the immobilized solution to immobilized carrier to solidify the immobilized carrier. The solidified immobilized carrier which comprises one kind of or various kinds of probiotics strains is washed by sterile water and then is sealed to be cultured for 8-72 hours at 10-45 DEG C. The produced immobilized probiotics do not need to be separated and frozen to dry, can be taken directly as probiotics and can be put in other food to take. If being sealed to store at normal temperature for a long time, the immobilized probiotics can grow without stop.
Description
(1) technical field
The present invention relates to a kind of production method of active probiotic, specifically belong to a kind of production method of edible immobilized active probiotics.
(2) background technology
Probiotic bacterium (Probiotics) is that a class is improved human intestine's microecological balance, the useful microorganism that HUMAN HEALTH is had opsonization, they mainly are to play the effect that promotes health by the microecological balance of safeguarding enteron aisle, and its physiologic function obtains whole world scientific and technological circle and generally admits.The whole world is delivered about the paper of research probiotic bacterium extremely abundant every year, summarizes to get up the effect that probiotic bacterium has following several respects:
(1) improve immunizing power: bifidus bacillus and Lactobacterium acidophilum can produce a large amount of immunoglobulin (Ig)s.Thereby activate the immunocyte of body, make external bacterium be difficult to retain in the body, reach immunoregulation effect.(2) improve gastrointestinal function: bifidus bacillus and Lactobacterium acidophilum form the protection barrier on the intestinal mucosa surface; resist the invasion and attack destruction of harmful bacterium to intestinal mucosa; can produce acidic substance 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 microflora.(3) trophism: bifidus bacillus and Lactobacterium acidophilum can produce lactic acid and acetic acid after fermenting in human intestinal, can improve the utilization ratio of calcium, phosphorus, iron, promote the absorption of iron and vitamins D, produce vitamin K and vitamins B, produce the Sumylact L lactose that helps digest.(4) logical intestines profit just: bifidus bacillus and Lactobacterium acidophilum can produce a large amount of acidic substance such as short chain fatty acid in intestines, can stimulate intestines peristalsis, increase ight soil humidity, play the effect of defaecation.(5) recover because of the normal microorganism species of ruined enteron aisle behind the use microbiotic: microbiotic is the important factor that causes alteration of intestinal flora.Microbiotic has also been killed a lot of probioticss in the kill harmful bacterium, in time replenish bifidus bacillus and Lactobacterium acidophilum, is very important to helping enteron aisle recovery colony balance, raising dietetic alimentation ability.(6) reducing cholesterol, prevent arteriosclerosis.(7) suppress endotoxic generation, delay senility.(8) generation of carcinogen in the inhibition enteron aisle, 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.At present, though probiotic products is extremely abundant on the market,, probiotic products has only liquid probiotic bacterium and two kinds of forms of probiotic bacterium lyophilized powder.
(3) summary of the invention
The object of the present invention is to provide a kind of production method of edible immobilized active probiotics, it comprises bifidumbacterium bifidum, Lactobacterium acidophilum, thermophilus streptococcus, lactobacillus delbruockii subspecies bulgaricus, bifidobacterium adolescentis, bifidus longum bb, bifidobacterium breve, bifidobacteria infantis, lactobacillus rhamnosus, lactobacterium casei, plant lactobacillus and animal bifid bar.
The object of the present invention is achieved like this:
(1) at first distinguishes the culture test tube slant strains, cultivated 12-72 hour, insert then in the triangular flask liquid nutrient medium and cultivate respectively or mixed culture, cultivated 8-72 hour for 20 ℃-45 ℃, carry out enlarged culturing more step by step, and then carry out ferment tank and cultivate, cultivated 8-72 hour for 20 ℃-45 ℃, make that total amount of probiotics reaches 1.0 * 10 in the fermented liquid
4-1.0 * 10
9/ mL, this nutrient solution is as seed liquor;
(2) will cultivate sophisticated seed liquor, be that the fixation support solution that contains suitable probiotic bacterium growth and breeding that it is good that 0.01%-20.0% adds the sterilization cooling is in the A liquid by concentration, thoroughly mixes and make fixation support solution B liquid; The compound method of described fixation support solution A liquid: take by weighing sodium alginate 0.1-200g, peptone 0.1-100.0 gram, whey powder 1.0-5.0 gram, Oligomeric maltose 1.0-50.0 gram, honey 1.0-50.0 gram, extractum carnis 0.1-10.0 gram, yeast extract paste 0.1-10.0 gram, Trisodium Citrate 0.1-10.0 gram, sodium-acetate 0.1-5.5 gram, dipotassium hydrogen phosphate 0.1-2.5 gram, sal epsom 0.01-0.75 gram, concentration is the 5-200mL of Tomato juice of 2.0%-25%, concentration is the Radix Dauci Sativae juice 5-200mL of 2.0%-25%, concentration is soaked juice 5-200mL for the 2.0%-25% soybean, add water and be settled to 1000 milliliters, thoroughly dissolving, pH5.0-7.0, sterilized 15 minutes, and be cooled to below 37 ℃ standby for 115 ℃.
(3) the fixation support solution B liquid that will contain the profitable probliotics seed pours in apparatus for making pearl ball or the syringe, B liquid is gently splashed in the fixation support solution C liquid, to reach B liquid and C liquid is 1: 1～1: 8 ratio, curing through 2～8 hours makes it to solidify, pull carrier ball sterilized water washed twice then out, place 10 ℃ of-45 ℃ of airtight cultivations 8-72 hour; The compound method of described fixation support solution C liquid: take by weighing calcium chloride 1.0-200.0g or Repone K 1.0-200.0g, add 1000mL water, stir, thoroughly dissolving, 115 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃.
The present invention also has some technical characterictics like this:
Another method of step (3) is that fixation support solution B liquid is put into volume is 0.01cm
3-1.0m
3In the container, the degree of depth 0.1～10cm of B liquid poured fixation support solution C liquid in the B liquid by 1: 1～1: 8 then gently, the curing through 2～8 hours, and that the immobilization probiotic bacterium can be made into is spherical, thread, column, strip, bulk, film like.
The collocation method of fixation support solution A liquid, described sodium alginate 0.1-200g substitutes with carrageenin 0.1-200g or carrageen 0.1-200g.Sodium alginate, kappa-carrageenan can use separately, but also both are mixed in proportion use,
The compound method of fixation support solution C liquid, described calcium chloride, Repone K can be prepared separately, but also mixed preparing, if sodium alginate, kappa-carrageenan use separately, then calcium chloride, Repone K are prepared separately.If sodium alginate, kappa-carrageenan mixed preparing, then calcium chloride, Repone K are with regard to mixed preparing.
The immobilized active probiotics that the present invention produced does not need to separate freeze-drying, can directly take as probiotic bacterium, but prolonged preservation under the normal temperature, viable count does not only reduce and can continue the breeding growth, and the bacterium number raises to some extent.
(4) embodiment
Embodiment one:
(1) with lactobacterium casei test tube slant bacterial classification, cultivated 12-72 hour, insert then in the triangular flask liquid nutrient medium and cultivate, cultivated 20 hours for 37 ℃, carry out enlarged culturing more step by step, and then carry out ferment tank and cultivate, cultivated 20 hours for 37 ℃, make that total count reaches 1.0 * 10 in the fermented liquid
9/ mL, this nutrient solution is as seed liquor;
(2) will cultivate sophisticated seed liquor, and add sterilization by 3.0% and cool off in the good sodium alginate fixation support solution (A liquid), and thoroughly mix and make (B liquid).
(3) then fixation support solution (B liquid) is splashed in 4% calcium chloride (C liquid), make it to solidify.The fixation support that contains lactobacterium casei after solidifying washs with sterilized water, places 10 ℃ of-45 ℃ of airtight cultivations 8-72 hour then.
(4) preparation of fixation support solution (A liquid): take by weighing sodium alginate 30.0g, peptone 5.0 grams, whey powder 5.0 grams, Oligomeric maltose 10.0 grams, honey 20.0 grams, extractum carnis 5.0 grams, yeast extract paste 2.0 grams, Trisodium Citrate 1.0 grams, sodium-acetate 1.0 grams, dipotassium hydrogen phosphate 1.0 grams, sal epsom 0.5 gram, concentration is that 10% the 50mL of Tomato juice, concentration are that 10% Radix Dauci Sativae juice 50mL, 5% soybean are soaked juice 50mL, Jia Shui and be settled to 1000 milliliters, thoroughly dissolving, pH5.0-7.0, sterilized 15 minutes, and be cooled to below 37 ℃ standby for 115 ℃.
(5) preparation of calcium chloride solid solution (C liquid): take by weighing calcium chloride 40.0g, add 1000mL water, stir, thoroughly dissolving, then, 115 ℃ of temperature were sterilized 15 minutes, were cooled to 25 ℃, and this solution is calcium chloride solid solution (C liquid).
(6) B liquid is poured in the syringe, gently splash in the calcium chloride solid solution (C liquid), the ratio of B liquid and calcium chloride solid solution (C liquid) was by 1: 4, and carrier ball sterilized water washed twice is pulled in the curing through 2 hours out, places 37 ℃ of airtight cultivations 8-72 hour then.Packing gets final product.
Embodiment two:
(1) with lactobacterium casei test tube slant bacterial classification, cultivated 12-72 hour, insert then in the triangular flask liquid nutrient medium and cultivate, cultivated 20 hours for 37 ℃, carry out enlarged culturing more step by step, and then carry out ferment tank and cultivate, cultivated 20 hours for 37 ℃, make that total count reaches 1.0 * 10 in the fermented liquid
9/ mL, this nutrient solution is as seed liquor;
(2) will cultivate sophisticated seed liquor, and add sterilization by 2.0% and cool off in the good kappa-carrageenan fixation support solution (A liquid), and thoroughly mix and make (B liquid).
(3) then fixation support solution (B liquid) is splashed in 4% Repone K (C liquid), make it to solidify.The fixation support that contains lactobacterium casei after solidifying washs with sterilized water, places 37 ℃ of airtight cultivations 20 hours then.
(4) preparation of fixation support solution (A liquid): take by weighing kappa-carrageenan 20.0 grams, peptone 5.0 grams, whey powder 5.0 grams, Oligomeric maltose 10.0 grams, honey 20.0 grams, extractum carnis 5.0 grams, yeast extract paste 2.0 grams, Trisodium Citrate 1.0 grams, sodium-acetate 1.0 grams, dipotassium hydrogen phosphate 1.0 grams, sal epsom 0.5 gram, concentration is that 10% the 50mL of Tomato juice, concentration are that 10% Radix Dauci Sativae juice 50mL, 5% soybean are soaked juice 50mL, Jia Shui and be settled to 1000 milliliters, thoroughly dissolving, pH5.0-7.0, sterilized 15 minutes, and be cooled to below 37 ℃ standby for 115 ℃.
(5) preparation of Repone K solid solution (C liquid): take by weighing Repone K 40.0g, add 1000mL water, stir, thoroughly dissolving, then, 115 ℃ of temperature were sterilized 15 minutes, were cooled to 25 ℃, and this solution is Repone K solid solution (C liquid).
(6) B liquid is poured in the syringe, gently splash in the Repone K solid solution (C liquid), the ratio of B liquid and Repone K solid solution (C liquid) was by 1: 4, and carrier ball sterilized water washed twice is pulled in the curing through 2 hours out, places 37 ℃ of airtight cultivations 8-72 hour then.Packing gets final product.
Embodiment three:
(1) with lactobacillus rhamnosus, Lactobacterium acidophilum test tube slant bacterial classification, cultivated 12-72 hour, insert respectively in the triangular flask liquid nutrient medium then and cultivate, cultivated 20 hours for 37 ℃, carry out enlarged culturing more step by step, and then carry out ferment tank and cultivate, cultivated 20 hours for 37 ℃, make that total count reaches 1.0 * 10 in the fermented liquid
9/ mL, this nutrient solution mix two liquid equal-volumes as seed liquor;
(2) will cultivate sophisticated seed liquor, and add sterilization by 2.0% and cool off in good sodium alginate, the kappa-carrageenan fixation support mixing solutions (A liquid), and thoroughly mix and make (B liquid).
(3) fixation support solution (B liquid) is splashed in 4% calcium chloride, 4% Repone K (C liquid) mixed solution then, make it to solidify.The fixation support that contains lactobacterium casei after solidifying washs with sterilized water, places 10 ℃ of-45 ℃ of airtight cultivations 8-72 hour then.
(4) preparation of fixation support solution (A liquid): take by weighing sodium alginate 30.0 grams, kappa-carrageenan 20.0 grams, peptone 5.0 grams, whey powder 5.0 grams, Oligomeric maltose 10.0 grams, honey 20.0 grams, extractum carnis 5.0 grams, yeast extract paste 2.0 grams, Trisodium Citrate 1.0 grams, sodium-acetate 1.0 grams, dipotassium hydrogen phosphate 1.0 grams, sal epsom 0.5 gram, concentration is 10% the 50mL of Tomato juice, concentration is 10% Radix Dauci Sativae juice 50mL, 5% soybean is soaked juice 50mL, add water and be settled to 1000 milliliters, thoroughly dissolving, pH5.0-7.0, sterilized 15 minutes, and be cooled to below 37 ℃ standby for 115 ℃.
(5) preparation of calcium chloride-Repone K solid solution (C liquid): take by weighing calcium chloride 40.0 grams, Repone K 40.0 grams, add 1000mL water, stir, thoroughly dissolving, then, 115 ℃ of temperature were sterilized 15 minutes, be cooled to 25 ℃, this solution is calcium chloride-Repone K solid solution (C liquid).
(6) B liquid is poured in the syringe, gently splash in calcium chloride-Repone K solid solution (C liquid), the ratio of B liquid and calcium chloride-Repone K solid solution (C liquid) was pressed 1: 4, the curing through 2 hours, pull carrier ball sterilized water washed twice out, place 37 ℃ of airtight cultivations 8-72 hour then.Packing gets final product.
Claims (3)
1. the production method of an edible immobilized active probiotics, described probiotic bacterium comprises: bifidumbacterium bifidum, Lactobacterium acidophilum, thermophilus streptococcus, lactobacillus delbruockii subspecies bulgaricus, bifidobacterium adolescentis, bifidus longum bb, bifidobacterium breve, bifidobacteria infantis, lactobacillus rhamnosus, lactobacterium casei, plant lactobacillus and animal bifidobacteria
Step is as follows:
(1) at first distinguishes the culture test tube slant strains, cultivated 12-72 hour, insert then in the triangular flask liquid nutrient medium and cultivate respectively or mixed culture, cultivated 8-72 hour for 20 ℃-45 ℃, carry out enlarged culturing more step by step, and then carry out ferment tank and cultivate, cultivated 8-72 hour for 20 ℃-45 ℃, make that total amount of probiotics reaches 1.0 * 10 in the fermented liquid
4-1.0 * 10
9/ mL, this nutrient solution is as seed liquor;
(2) will cultivate sophisticated seed liquor, be that 0.01%-20.0% adds good the containing in the fixation support solution A liquid that is fit to the probiotic bacterium growth and breeding of sterilization cooling by concentration, thoroughly mixes and make fixation support solution B liquid; Described A liquid making method, it is characterized in that: take by weighing sodium alginate 0.1-200g, peptone 0.1-100.0 gram, whey powder 1.0-5.0 gram, Oligomeric maltose 1.0-50.0 gram, honey 1.0-50.0 gram, extractum carnis 0.1-10.0 gram, yeast extract paste 0.1-10.0 gram, Trisodium Citrate 0.1-10.0 gram, sodium-acetate 0.1-5.5 gram, dipotassium hydrogen phosphate 0.1-2.5 gram, sal epsom 0.01-0.75 gram, concentration is the 5-200mL of Tomato juice of 2.0%-25%, concentration is the Radix Dauci Sativae juice 5-200mL of 2.0%-25%, concentration is soaked juice 5-200mL for the 2.0%-25% soybean, add water and be settled to 1000 milliliters, thoroughly dissolving, pH5.0-7.0, sterilized 15 minutes, and be cooled to below 37 ℃ standby for 115 ℃;
(3) the fixation support solution B liquid that will contain the profitable probliotics seed pours in apparatus for making pearl ball or the syringe, B liquid is gently splashed in the fixation support solution C liquid, to reach B liquid and C liquid is 1: 1～1: 8 ratio, curing through 2～8 hours makes it to solidify, pull carrier ball sterilized water washed twice then out, place 10 ℃ of-45 ℃ of airtight cultivations 8-72 hour; The compound method of described C liquid is characterized in that: takes by weighing calcium chloride 1.0-200.0g or Repone K 1.0-200.0g, adds 1000mL water, stir, and thoroughly dissolving, 115 ℃ of temperature were sterilized 15 minutes, were cooled to 10 ℃-35 ℃.
2. the production method of a kind of edible immobilized active probiotics according to claim 1 is characterized in that the another kind of method of described step (3) is that fixation support solution B liquid is put into volume is 0.01cm
3-1.0m
3In the container, the degree of depth 0.1～10cm of B liquid poured fixation support solution C liquid in the B liquid by 1: 1～1: 8 then gently, the curing through 2～8 hours, and that the immobilization probiotic bacterium can be made into is spherical, thread, column, strip, bulk or film like.
3. the production method of a kind of edible immobilized active probiotics according to claim 1 is characterized in that sodium alginate 0.1-200g substitutes with carrageenin 0.1-200g or carrageen 0.1-200g in the described step.
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