CN112244153A - Preparation method of double-layer coated lactic acid bacteria microcapsule - Google Patents
Preparation method of double-layer coated lactic acid bacteria microcapsule Download PDFInfo
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- CN112244153A CN112244153A CN202010890657.5A CN202010890657A CN112244153A CN 112244153 A CN112244153 A CN 112244153A CN 202010890657 A CN202010890657 A CN 202010890657A CN 112244153 A CN112244153 A CN 112244153A
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- acid bacteria
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- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 88
- 241000894006 Bacteria Species 0.000 title claims abstract description 50
- 239000004310 lactic acid Substances 0.000 title claims abstract description 44
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 44
- 239000003094 microcapsule Substances 0.000 title claims abstract description 31
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 238000000576 coating method Methods 0.000 claims abstract description 50
- 239000011248 coating agent Substances 0.000 claims abstract description 49
- 241000186660 Lactobacillus Species 0.000 claims abstract description 46
- 229940039696 lactobacillus Drugs 0.000 claims abstract description 46
- 239000000463 material Substances 0.000 claims abstract description 33
- 239000000843 powder Substances 0.000 claims abstract description 11
- 210000001035 gastrointestinal tract Anatomy 0.000 claims abstract description 9
- 239000000243 solution Substances 0.000 claims description 44
- 238000005507 spraying Methods 0.000 claims description 42
- 238000005469 granulation Methods 0.000 claims description 21
- 230000003179 granulation Effects 0.000 claims description 21
- 230000001580 bacterial 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 12
- 239000001110 calcium chloride Substances 0.000 claims description 12
- 235000011148 calcium chloride Nutrition 0.000 claims description 12
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 12
- 241001465754 Metazoa Species 0.000 claims description 11
- 239000002245 particle Substances 0.000 claims description 11
- 239000003921 oil Substances 0.000 claims description 9
- 235000019198 oils Nutrition 0.000 claims description 9
- 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 8
- 235000010469 Glycine max Nutrition 0.000 claims description 8
- 244000068988 Glycine max Species 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- 235000020183 skimmed milk Nutrition 0.000 claims description 8
- 235000010413 sodium alginate Nutrition 0.000 claims description 8
- 239000000661 sodium alginate Substances 0.000 claims description 8
- 229940005550 sodium alginate Drugs 0.000 claims description 8
- 240000008415 Lactuca sativa Species 0.000 claims description 7
- 108010073771 Soybean Proteins Proteins 0.000 claims description 7
- 238000000034 method Methods 0.000 claims description 7
- 235000012045 salad Nutrition 0.000 claims description 7
- 239000010802 sludge Substances 0.000 claims description 7
- 235000019710 soybean protein Nutrition 0.000 claims description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- 235000010418 carrageenan Nutrition 0.000 claims description 6
- 239000000679 carrageenan Substances 0.000 claims description 6
- 229920001525 carrageenan Polymers 0.000 claims description 6
- 229940113118 carrageenan Drugs 0.000 claims description 6
- 230000008569 process Effects 0.000 claims description 6
- 239000011734 sodium Substances 0.000 claims description 6
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 claims description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 5
- 238000000889 atomisation Methods 0.000 claims description 5
- 238000007865 diluting Methods 0.000 claims description 5
- 238000009835 boiling Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
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- 235000015112 vegetable and seed oil Nutrition 0.000 claims description 4
- 239000008158 vegetable oil Substances 0.000 claims description 4
- 229920002472 Starch Polymers 0.000 claims description 3
- 235000010489 acacia gum Nutrition 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
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- 244000215068 Acacia senegal Species 0.000 claims description 2
- 229920000084 Gum arabic Polymers 0.000 claims description 2
- 235000019484 Rapeseed oil Nutrition 0.000 claims description 2
- 239000000205 acacia gum Substances 0.000 claims description 2
- 235000005687 corn oil Nutrition 0.000 claims description 2
- 239000002285 corn oil Substances 0.000 claims description 2
- 235000012343 cottonseed oil Nutrition 0.000 claims description 2
- 239000002385 cottonseed oil Substances 0.000 claims description 2
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 2
- 235000021388 linseed oil Nutrition 0.000 claims description 2
- 239000000944 linseed oil Substances 0.000 claims description 2
- 229940099596 manganese sulfate Drugs 0.000 claims description 2
- 235000007079 manganese sulphate Nutrition 0.000 claims description 2
- 239000011702 manganese sulphate Substances 0.000 claims description 2
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 2
- 239000001267 polyvinylpyrrolidone Substances 0.000 claims description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 2
- 238000007789 sealing Methods 0.000 claims description 2
- 238000000926 separation method Methods 0.000 claims description 2
- 229940073490 sodium glutamate Drugs 0.000 claims description 2
- 235000020238 sunflower seed Nutrition 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 3
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 235000013861 fat-free Nutrition 0.000 abstract 1
- 239000008267 milk Substances 0.000 abstract 1
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- 244000144972 livestock Species 0.000 description 8
- 238000009395 breeding Methods 0.000 description 6
- 230000001488 breeding effect Effects 0.000 description 6
- 239000007921 spray Substances 0.000 description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical group O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 4
- 244000144977 poultry Species 0.000 description 4
- 229930006000 Sucrose Natural products 0.000 description 3
- 229940124350 antibacterial drug Drugs 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000005243 fluidization Methods 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 239000000047 product Substances 0.000 description 3
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- 229960004793 sucrose Drugs 0.000 description 3
- 229920002774 Maltodextrin Polymers 0.000 description 2
- 239000005913 Maltodextrin Substances 0.000 description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 239000003674 animal food additive Substances 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003640 drug residue Substances 0.000 description 2
- 229940035034 maltodextrin Drugs 0.000 description 2
- 229910021645 metal ion Inorganic materials 0.000 description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 2
- 239000008108 microcrystalline cellulose Substances 0.000 description 2
- 239000001763 2-hydroxyethyl(trimethyl)azanium Substances 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 235000019743 Choline chloride Nutrition 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 235000007685 Pleurotus columbinus Nutrition 0.000 description 1
- 240000001462 Pleurotus ostreatus Species 0.000 description 1
- 235000001603 Pleurotus ostreatus Nutrition 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229960003178 choline chloride Drugs 0.000 description 1
- SGMZJAMFUVOLNK-UHFFFAOYSA-M choline chloride Chemical compound [Cl-].C[N+](C)(C)CCO SGMZJAMFUVOLNK-UHFFFAOYSA-M 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
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- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000006479 redox reaction Methods 0.000 description 1
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- 239000001632 sodium acetate Substances 0.000 description 1
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- 239000004455 soybean meal Substances 0.000 description 1
- 235000015193 tomato juice Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
- A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
Abstract
The invention discloses a preparation method of a double-layer coated lactobacillus microcapsule, which comprises the following steps: (1) preparing a lactic acid bacteria solution; (2) vulcanizing and granulating; (3) primary coating; (4) and secondary coating, wherein the formula of the microcapsule coating wall material is optimized, so that the resistance of the freeze-dried bacterium powder is improved, the non-fat dry milk powder and the like are prevented from being decomposed after entering the intestinal tract, nutrition is provided for the fixed value of the lactic acid bacteria, and the survival rate of the lactic acid bacteria is greatly improved by the secondary coating process.
Description
Technical Field
The invention relates to the technical field of lactobacillus coating, in particular to a preparation method of a double-layer coated lactobacillus microcapsule.
Background
For a long time, the application of the antibiotic feed additive in the livestock aquaculture industry has achieved good results, and the development of the livestock aquaculture industry is greatly promoted, however, the addition of the antibacterial drugs in the premix and the blind abuse of the antibacterial drugs in the breeding process cause a series of problems: firstly, the feed and the breeding industry seriously depend on antibacterial drugs, so that the drug resistance of pathogenic bacteria and viruses in the animal body and the breeding environment is intensified, the drug effect is reduced, the breeding cost is increased, the ecological environment in the animal body and the animal body is damaged, and the difficulty in preventing and controlling the animal epidemic diseases is increased; secondly, the problems of drug residues in livestock products are serious, the product quality and safety are reduced, the drug residue problem causes frequent obstruction of the livestock product export in China, economic loss is heavy, and the market competitiveness and the trust of consumers are reduced; thirdly, the environment pollution is serious, a large amount of drug-resistant bacteria and harmful livestock and poultry excrement exist in the breeding environment, the ecological environment is worsened, the human health is threatened, and the like, the lactobacillus type microbial feed additive is an important one, the lactobacillus is a resident beneficial bacterium in the animal body, and can resist pathogenic microorganisms and adjust the balance of animal digestive tract microflora; activating immune system and enhancing immunity; in addition, the lactobacillus can also prevent diarrhea of livestock and poultry, improve the weight gain of livestock and poultry, and play an important physiological function in intestinal tracts.
However, since most lactic acid bacteria do not produce spores, have poor stress resistance, and are easily inactivated, their use value is seriously affected, for example, the pH of gastric juice is usually below 2.5, and the acidity is such that most of the ingested lactic acid bacteria are destroyed, and only about one part per million of lactic acid bacteria can reach the large intestine alive. When lactic acid bacteria are added to feed for pelleting, the usual pelleting temperature of the feed is 80-C, and the lactobacillus can be lost by 70-80% in only 5 minutes. In addition, in feed production and livestock and poultry breeding, in order to be convenient to use, lactic acid bacteria are usually directly added into complete feed or additive premix, and because both of the lactic acid bacteria and the additive premix have a large amount of trace element metal ions (such as Cu2+, Fe2+, Mn2+, Zn2+ and Ca2+), vitamins (such as choline chloride) and antibiotics, the survival and the activity of the lactic acid bacteria can be influenced in a series of oxidation-reduction reactions, pH value changes, the survival microenvironment of the lactic acid bacteria is changed, and the like.
Disclosure of Invention
The invention aims to design a preparation method of a double-layer coated lactic acid bacteria microcapsule to solve the problems in the background technology.
In order to achieve the purpose, the invention provides the following technical scheme:
a preparation method of a double-layer coated lactobacillus microcapsule comprises the following steps:
(1) preparation of lactic acid bacteria solution:
inoculating lactic acid bacteria separated from intestinal tracts of healthy animals into a sterilized culture medium for culture, then carrying out centrifugal separation on the obtained lactic acid bacteria culture solution, diluting the separated precipitate, namely bacterial sludge, into a mixed solution containing 109-1011 cfu/ml of viable lactic acid bacteria by using sterilized normal saline, and adding skimmed milk powder accounting for 4-9% of the total weight of the precipitate, namely bacterial sludge, to obtain a lactic acid bacteria solution;
(2) and (3) vulcanizing and granulating:
spraying the lactobacillus solution prepared in the step (1) in a rotating pelleting pot of a fluidized granulation coating dryer filled with pelleting auxiliary materials in a boiling state, stopping spraying the lactobacillus solution after l3-26min, and continuously boiling and rotating the pelleting pot for 6-8 min to obtain spherical lactobacillus particles with the diameter of 1.1-1.8 mm, wherein the content of lactobacillus in the spherical lactobacillus particles accounts for 22-28% of the total weight of the spherical lactobacillus particles;
the process conditions of the fluidized granulation coating dryer used are as follows: the consumption of compressed air is 0.25-0.5 m3/min, the pressure of air sealing is 0.14-0.17 Mpa, the atomization pressure is 0.17-0.20 Mpa, the current pressure is 0.25-0.51 Mpa, the material spraying speed is 5.1-13.5 ml/min, the inlet temperature of the sprayed material is 25.5-92.5 ℃, and the outlet temperature is normal temperature and normal pressure;
(3) primary coating:
putting the spherical lactic acid bacteria particles obtained in the step (2) into a coating pan in a fluidized granulation coating dryer, simultaneously spraying 1.2-5.0 wt% of wall material solution and 1.2-3.0 wt% of CaCl2 solution from an inlet of the coating pan, wherein the temperature of a spraying inlet is 68-72 ℃, the spraying speed is 5-18 ml/min, and after 15-27 minutes, obtaining coated lactic acid bacteria microcapsules with the diameter of 1.8-2.7 mm through an outlet in the fluidized granulation coating dryer, and the outlet temperature is 35-46 ℃.
(4) Secondary coating:
putting the primary microcapsule obtained in the step (3) into a coating pan in a fluidized granulation coating dryer again, simultaneously spraying 1.1-5.1 wt% of wall material solution and 1.1-3.1 wt% of CaCl2 solution from an inlet of the coating pan, wherein the temperature of a spraying material inlet is 65.5-74.5 ℃, the spraying speed is 2-20 ml/min, and after 12-28 minutes, obtaining a double-layer coated lactic acid bacteria microcapsule with the diameter of 2.2-3.0 mm through an outlet in the fluidized granulation coating dryer, wherein the outlet temperature is 36-47 ℃;
the wall material is one or a mixture of two or more of sodium alginate, CaCl2 and carrageenan, soybean protein isolate and water-insoluble vegetable oil, and other wall material components: 1-4 g of sodium glutamate, 0.2-1.8 g of manganese sulfate, 1-1.7 g of glycerol and 300.1-0.8 g of polyvinylpyrrolidone K; wherein the inner layer is coated, and microcapsule wall material including gum arabic and sodium carboxymethyl starch (CMS-Na) is added into conventional lyophilized protectant.
Preferably, the water-insoluble vegetable oil is linseed oil, cottonseed oil, rapeseed oil, soybean salad oil, sunflower seed oil or corn oil.
Compared with the prior art, the invention has the beneficial effects that: by optimizing the formula of the microcapsule coating wall material and adding microcapsule wall materials, namely, Arabic gum and sodium carboxymethyl starch (CMS-Na) into a conventional freeze-drying protective agent, thalli, skimmed milk powder and cane sugar form a relatively stable spatial structure through microencapsulation, so that the resistance of freeze-dried bacterial powder is improved, the skimmed milk powder and the like are prevented from being decomposed after entering an intestinal tract, and nutrition is provided for lactobacillus in a fixed value; the outer coating increases the barrier property, firmness and elasticity of the microcapsule, prevents the destruction of metal ions, gastric acid and bile salt in oxygen and feed, and the secondary coating enables the thallus yield to reach more than 90 percent, the thallus microcapsule coating efficiency to reach more than 75 percent, the thallus coating yield to reach more than 85 percent, the quality is stable, the quality guarantee period is long, and the room temperature can reach 12 months.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a flow chart of the preparation method of the double-layer coated lactic acid bacteria microcapsule of the present invention.
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the accompanying drawings, and obviously, the description is only a part of the embodiments of the present invention, not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Example 1
Using a culture medium comprising components A, B, C and D, wherein component A is one or more of beef extract or peptone, yeast extract, skimmed milk/powder, soybean peptide, tryptone, and soybean meal; the component B is one or a mixture of more of lactose, glucose, tomato juice and malt extract; the component C is one or more of Tween-80, cysteine, Sucus Dauci Sativae, rhizoma Solani Tuber osi juice, herba Alii Fistulosi juice, and Pleurotus Ostreatus juice; the component D is one or a mixture of more of NaCI, KH2P04, diammonium citrate, sodium acetate and MgS04, the culture medium is sterilized at 121 ℃ for 15-30 min, and the culture medium is cooled to room temperature.
Separating a lactic acid bacteria strain from intestinal tracts of healthy animals, inoculating the lactic acid bacteria strain into the culture medium, and culturing at 40 ℃ for 12 hours; centrifuging the culture solution for 30min at 4000r/min to obtain lactobacillus bacterial sludge;
diluting the lactobacillus bacterial mud with sterilized normal saline, and adding skimmed milk powder with the weight of 5% of the total weight of the lactobacillus bacterial mud to obtain lactobacillus concentrated solution with viable bacteria concentration of 1010 cfu/ml.
An FLP type fluidization granulation coating dryer is adopted, and the ratio of sucrose: maltodextrin, i.e.: microcrystalline cellulose: 5: 4: 1 (weight ratio), putting 2kg of pelleting auxiliary materials into a pelleting pot, and spraying the lactobacillus concentrated solution, wherein the weight ratio of the pelleting auxiliary materials to the lactobacillus concentrated solution is 1: 2 the process conditions of the fluidized granulation coating dryer used are as follows: the consumption of compressed air is 0.4m3/min, the pressure of air seal is adjusted to 0.16Mpa, the atomization pressure is 0.18Mpa, the current is adjusted to 0.35Mpa, the spraying feeding speed is 10ml/min, and the spraying inlet temperature and the spraying outlet temperature are both normal temperature; and (3) starting the FLP type fluidized granulation coating dryer, observing the pelleting condition through an equipment window, and preparing spherical lactic acid bacteria particles with the diameter of 1.1-1.8 mm after 16 min.
And starting a coating device to carry out primary coating, spraying wall material solution of 2 wt% of sodium alginate, 1 wt% of carrageenan, 0.5 wt% of soybean salad oil and 0.4 wt% of soybean protein isolate at the inlet temperature of 72 ℃ and the outlet temperature of 45 ℃, and spraying 2 wt% of CaCl2 solution from another nozzle at the same time, wherein the feeding speeds are 8ml/min and 4ml/min respectively, so as to prepare the coated lactic acid bacteria microcapsule with the diameter of 2.1-2.9 mm, and the content of lactic acid bacteria accounts for 22% of the total weight of the microcapsule.
And (3) secondary coating, namely spraying 1.8 wt% of sodium alginate, 1 wt% of carrageenan, 0.5 wt% of soybean salad oil and 0.4 wt% of wall material solution of soybean protein isolate at the inlet temperature of 70 ℃ and the outlet temperature of 44 ℃, and spraying 2 wt% of CaCl2 solution from another spray head at the same time, wherein the feeding speeds are 7ml/min and 4ml/min respectively, so as to prepare the double-layer coated lactobacillus microcapsule with the diameter of 2.2-3.0 mm.
Example 2
Separating a lactic acid bacterium from the intestinal tract of a healthy animal, inoculating the lactic acid bacterium into the culture medium described in example 1, and culturing at 25-42 ℃ for 12 hours; centrifuging the culture solution for 30min at 4000r/min to obtain lactobacillus bacterial sludge; diluting the lactobacillus bacterial mud with sterilized normal saline, and adding skimmed milk powder accounting for 3% of the total weight of the lactobacillus bacterial mud to obtain lactobacillus concentrated solution with viable bacteria concentration of 109 cfu/mL.
And (3) adopting an FLP type fluidization granulation coating dryer to mix the microcrystalline cellulose: sucrose: corn starch: 2: 3: 5 (weight ratio), putting 2kg of pelleting auxiliary materials into a pelleting pot, and spraying the lactobacillus concentrated solution, wherein the weight ratio of the pelleting auxiliary materials to the lactobacillus concentrated solution is 1: 2, spraying twice; the process conditions of the fluidized granulation coating dryer used are as follows: the consumption of compressed air is 0.25m3/min, the pressure of air seal is 0.14Mpa, the atomization pressure is 0.17Mpa,
regulating current pressure to 0.25Mpa, spraying at feeding speed of 5.1ml/min, spraying inlet air temperature of 50 deg.C, and outlet temperature of normal temperature; and (3) starting the FLP type fluidized granulation coating dryer, observing the pelleting condition through an equipment window, and preparing spherical lactic acid bacteria particles with the diameter of 1.1-1.8 mm after 10 min.
Starting a coating device, spraying a wall material solution of 3 wt% of sodium alginate, 0.6 wt% of soybean salad oil and 0.5 wt% of soybean protein isolate at the inlet temperature of 65 ℃ and the outlet temperature of 30 ℃, and simultaneously spraying a CaCl2 solution of 1 wt% from another spray head at feeding speeds of 4ml/in and 2ml/min respectively to prepare the coated lactobacillus microcapsule with the diameter of 2.1-2.9 mm, wherein the content of lactobacillus accounts for 25% of the total weight of the microcapsule.
And (3) secondary coating, namely spraying 1.7 wt% of sodium alginate, 1 wt% of carrageenan, 0.5 wt% of soybean salad oil and 0.4 wt% of wall material solution of soybean protein isolate at the inlet temperature of 68 ℃ and the outlet temperature of 43 ℃, and spraying 2 wt% of CaCl2 solution from another spray head at the same time, wherein the feeding speeds are 7ml/min and 4ml/min respectively, so as to prepare the double-layer coated lactobacillus microcapsule with the diameter of 2.2-3.0 mm.
Example 3
Separating a lactic acid bacterium from the intestinal tract of a healthy animal, inoculating the lactic acid bacterium into the culture medium described in example 1, and culturing at 25-42 ℃ for 12 hours; centrifuging the culture solution for 30min at 4000r/min to obtain lactobacillus bacterial sludge; diluting the lactobacillus bacterial mud with sterilized normal saline, and adding skimmed milk powder accounting for 10% of the total weight of the lactobacillus bacterial mud to obtain lactobacillus concentrated solution with viable bacteria concentration of 1011 cfu.
An FLP type fluidization granulation coating dryer is adopted, and the ratio of sucrose: maltodextrin, i.e.: protein sugar: 2: 3: 4 (weight ratio), putting 2kS of pelleting auxiliary materials into a pelleting pot, and spraying the lactobacillus concentrated solution, wherein the weight ratio of the pelleting auxiliary materials to the lactobacillus concentrated solution is 1: 3, carrying out three times of spraying; the process conditions of the fluidized granulation coating dryer used are as follows: the consumption of compressed air is 0.6m3/min, the pressure of air seal is adjusted to 0.20Mpa, the atomization pressure is 0.22Mpa, the current is adjusted to 0.6Mpa, the spraying feeding speed is 15mVmin, the spraying inlet air temperature is 95 ℃, and the outlet temperature is normal temperature; and (3) starting the FLP type fluidized granulation coating dryer, observing the pelleting condition through an equipment window, and preparing spherical lactic acid bacteria particles with the diameter of 1.1-1.9 mm after 30 min. And starting a coating device, spraying 5 wt% of sodium alginate wall material solution at an inlet temperature of 75 ℃ and an outlet temperature of 50' C, and spraying 3 wt% of CaCl2 solution from another spray head at feeding speeds of 20ml/min and 10mi/rain respectively to prepare the coated lactobacillus microcapsules with diameters of 2.1-2.9 mm, wherein the content of lactobacillus accounts for 30% of the total weight of the microcapsules.
And (3) secondary coating, namely spraying wall material solution of 1.6 wt% of sodium alginate, 1 wt% of carrageenan, 0.5 wt% of soybean salad oil and 0.4 wt% of soybean protein isolate at the inlet temperature of 66 ℃ and the outlet temperature of normal temperature, and spraying 2 wt% of CaCl2 solution from another spray head at the same time, wherein the feeding speeds are 6ml/min and 4ml/min respectively, so as to prepare the double-layer coated lactobacillus microcapsule with the diameter of 2.2-3.0 mm.
While the preferred embodiments of the present invention have been illustrated and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (2)
1. A preparation method of a double-layer coated lactic acid bacteria microcapsule is characterized by comprising the following steps:
(1) preparation of lactic acid bacteria solution:
inoculating lactic acid bacteria separated from intestinal tracts of healthy animals into a sterilized culture medium for culture, then carrying out centrifugal separation on the obtained lactic acid bacteria culture solution, diluting the separated precipitate, namely bacterial sludge, into a mixed solution containing 109-1011 cfu/ml of viable lactic acid bacteria by using sterilized normal saline, and adding skimmed milk powder accounting for 4-9% of the total weight of the precipitate, namely bacterial sludge, to obtain a lactic acid bacteria solution;
(2) and (3) vulcanizing and granulating:
spraying the lactobacillus solution prepared in the step (1) in a rotating pelleting pot of a fluidized granulation coating dryer filled with pelleting auxiliary materials in a boiling state, stopping spraying the lactobacillus solution after l3-26min, and continuously boiling and rotating the pelleting pot for 6-8 min to obtain spherical lactobacillus particles with the diameter of 1.1-1.8 mm, wherein the content of lactobacillus in the spherical lactobacillus particles accounts for 22-28% of the total weight of the spherical lactobacillus particles;
the process conditions of the fluidized granulation coating dryer used are as follows: the consumption of compressed air is 0.25-0.5 m3/min, the pressure of air sealing is 0.14-0.17 Mpa, the atomization pressure is 0.17-0.20 Mpa, the current pressure is 0.25-0.51 Mpa, the material spraying speed is 5.1-13.5 ml/min, the inlet temperature of the sprayed material is 25.5-92.5 ℃, and the outlet temperature is normal temperature and normal pressure;
(3) primary coating:
putting the spherical lactic acid bacteria particles obtained in the step (2) into a coating pan in a fluidized granulation coating dryer, simultaneously spraying 1.2-5.0 wt% of wall material solution and 1.2-3.0 wt% of CaCl2 solution from an inlet of the coating pan, wherein the temperature of a spraying inlet is 68-72 ℃, the spraying speed is 5-18 ml/min, and after 15-27 minutes, obtaining coated lactic acid bacteria microcapsules with the diameter of 1.8-2.7 mm through an outlet in the fluidized granulation coating dryer, and the outlet temperature is 35-46 ℃.
(4) Secondary coating:
putting the primary microcapsule obtained in the step (3) into a coating pan in a fluidized granulation coating dryer again, simultaneously spraying 1.1-5.1 wt% of wall material solution and 1.1-3.1 wt% of CaCl2 solution from an inlet of the coating pan, wherein the temperature of a spraying material inlet is 65.5-74.5 ℃, the spraying speed is 2-20 ml/min, and after 12-28 minutes, obtaining a double-layer coated lactic acid bacteria microcapsule with the diameter of 2.2-3.0 mm through an outlet in the fluidized granulation coating dryer, wherein the outlet temperature is 36-47 ℃;
the wall material is one or a mixture of two or more of sodium alginate, CaCl2 and carrageenan, soybean protein isolate and water-insoluble vegetable oil, and other wall material components: 1-4 g of sodium glutamate, 0.2-1.8 g of manganese sulfate, 1-1.7 g of glycerol and 300.1-0.8 g of polyvinylpyrrolidone K; wherein the inner layer is coated, and microcapsule wall material including gum arabic and sodium carboxymethyl starch (CMS-Na) is added into conventional lyophilized protectant.
2. The method for preparing a double-layer coated lactic acid bacteria microcapsule according to claim 1, wherein: the water-insoluble vegetable oil is linseed oil, cottonseed oil, rapeseed oil, soybean salad oil, sunflower seed oil or corn oil.
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CN1969889A (en) * | 2006-12-04 | 2007-05-30 | 济南赛拜斯生物工程有限公司 | Enteric-coated multilayer encapsulated probiotic microcapsule and preparation method thereof |
US20120282304A1 (en) * | 2010-11-02 | 2012-11-08 | Cell Biotech Co., Ltd. | Multi-coated lactic acid bacteria and preparing method thereof |
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CN1969889A (en) * | 2006-12-04 | 2007-05-30 | 济南赛拜斯生物工程有限公司 | Enteric-coated multilayer encapsulated probiotic microcapsule and preparation method thereof |
US20120282304A1 (en) * | 2010-11-02 | 2012-11-08 | Cell Biotech Co., Ltd. | Multi-coated lactic acid bacteria and preparing method thereof |
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