CN112544977B - Preparation method of multi-layer coated probiotic microcapsule - Google Patents
Preparation method of multi-layer coated probiotic microcapsule Download PDFInfo
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- CN112544977B CN112544977B CN202011208280.7A CN202011208280A CN112544977B CN 112544977 B CN112544977 B CN 112544977B CN 202011208280 A CN202011208280 A CN 202011208280A CN 112544977 B CN112544977 B CN 112544977B
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- 239000003094 microcapsule Substances 0.000 title claims abstract description 110
- 239000006041 probiotic Substances 0.000 title claims abstract description 45
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 45
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 238000000855 fermentation Methods 0.000 claims abstract description 25
- 230000004151 fermentation Effects 0.000 claims abstract description 25
- 239000011248 coating agent Substances 0.000 claims abstract description 16
- 238000000576 coating method Methods 0.000 claims abstract description 16
- 235000013305 food Nutrition 0.000 claims abstract description 5
- 239000010410 layer Substances 0.000 claims description 46
- 230000001580 bacterial effect Effects 0.000 claims description 45
- 239000010802 sludge Substances 0.000 claims description 44
- 241000194031 Enterococcus faecium Species 0.000 claims description 31
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 30
- 239000003223 protective agent Substances 0.000 claims description 30
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 28
- 239000000463 material Substances 0.000 claims description 26
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 22
- 241000894006 Bacteria Species 0.000 claims description 21
- 239000000243 solution Substances 0.000 claims description 20
- 238000000034 method Methods 0.000 claims description 18
- 229920001661 Chitosan Polymers 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 239000002356 single layer Substances 0.000 claims description 15
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 14
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 14
- 239000004310 lactic acid Substances 0.000 claims description 14
- 235000014655 lactic acid Nutrition 0.000 claims description 14
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 14
- 230000001954 sterilising effect Effects 0.000 claims description 14
- 229960000583 acetic acid Drugs 0.000 claims description 12
- 239000002702 enteric coating Substances 0.000 claims description 12
- 238000009505 enteric coating Methods 0.000 claims description 12
- 239000012362 glacial acetic acid Substances 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- 230000001502 supplementing effect Effects 0.000 claims description 12
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 11
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 10
- 239000004594 Masterbatch (MB) Substances 0.000 claims description 10
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 10
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 10
- 238000005469 granulation Methods 0.000 claims description 9
- 230000003179 granulation Effects 0.000 claims description 9
- 238000002156 mixing Methods 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
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 8
- NWGKJDSIEKMTRX-AAZCQSIUSA-N Sorbitan monooleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O NWGKJDSIEKMTRX-AAZCQSIUSA-N 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 239000007853 buffer solution Substances 0.000 claims description 8
- 239000003795 chemical substances by application Substances 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000005022 packaging material Substances 0.000 claims description 8
- 239000001632 sodium acetate Substances 0.000 claims description 8
- 235000017281 sodium acetate Nutrition 0.000 claims description 8
- 239000000661 sodium alginate Substances 0.000 claims description 8
- 235000010413 sodium alginate Nutrition 0.000 claims description 8
- 229940005550 sodium alginate Drugs 0.000 claims description 8
- 235000020183 skimmed milk Nutrition 0.000 claims description 6
- 238000005507 spraying Methods 0.000 claims description 6
- 239000004925 Acrylic resin Substances 0.000 claims description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 4
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 4
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 claims description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- 239000001768 carboxy methyl cellulose Substances 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims description 4
- 239000008120 corn starch Substances 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
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- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 4
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- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 claims description 4
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 claims description 4
- 238000005303 weighing Methods 0.000 claims description 4
- 239000000230 xanthan gum Substances 0.000 claims description 4
- 235000010493 xanthan gum Nutrition 0.000 claims description 4
- 229920001285 xanthan gum Polymers 0.000 claims description 4
- 229940082509 xanthan gum Drugs 0.000 claims description 4
- 241000186000 Bifidobacterium Species 0.000 claims description 2
- 241001478240 Coccus Species 0.000 claims description 2
- 241000194032 Enterococcus faecalis Species 0.000 claims description 2
- 241000186660 Lactobacillus Species 0.000 claims description 2
- 244000199866 Lactobacillus casei Species 0.000 claims description 2
- 235000013958 Lactobacillus casei Nutrition 0.000 claims description 2
- 241001147746 Lactobacillus delbrueckii subsp. lactis Species 0.000 claims description 2
- 241000186604 Lactobacillus reuteri Species 0.000 claims description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 2
- 229940032049 enterococcus faecalis Drugs 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims description 2
- 229940039696 lactobacillus Drugs 0.000 claims description 2
- 229940017800 lactobacillus casei Drugs 0.000 claims description 2
- 229940001882 lactobacillus reuteri Drugs 0.000 claims description 2
- 239000004816 latex Substances 0.000 claims description 2
- 229920000126 latex Polymers 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 238000007873 sieving Methods 0.000 claims description 2
- 230000002378 acidificating effect Effects 0.000 claims 1
- 239000000825 pharmaceutical preparation Substances 0.000 claims 1
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- 230000004083 survival effect Effects 0.000 abstract description 27
- 239000003814 drug Substances 0.000 abstract description 3
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 3
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- 230000000694 effects Effects 0.000 description 5
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- 108010010803 Gelatin Proteins 0.000 description 2
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- 238000005516 engineering process Methods 0.000 description 2
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- 229920000159 gelatin Polymers 0.000 description 2
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- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
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- 230000004048 modification Effects 0.000 description 2
- 238000004088 simulation Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
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- 239000003613 bile acid Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
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- 238000009776 industrial production Methods 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 229940127554 medical product Drugs 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000001819 pancreatic juice Anatomy 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000013441 quality evaluation Methods 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P20/00—Coating of foodstuffs; Coatings therefor; Making laminated, multi-layered, stuffed or hollow foodstuffs
- A23P20/10—Coating with edible coatings, e.g. with oils or fats
- A23P20/15—Apparatus or processes for coating with liquid or semi-liquid products
- A23P20/18—Apparatus or processes for coating with liquid or semi-liquid products by spray-coating, fluidised-bed coating or coating by casting
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/46—Ingredients of undetermined constitution or reaction products thereof, e.g. skin, bone, milk, cotton fibre, eggshell, oxgall or plant extracts
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5026—Organic macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
-
- A—HUMAN NECESSITIES
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- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/125—Casei
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/157—Lactis
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/173—Reuteri
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- Public Health (AREA)
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- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
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- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
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- Chemical Kinetics & Catalysis (AREA)
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- Biomedical Technology (AREA)
- Biotechnology (AREA)
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Abstract
The invention discloses a preparation method of a multi-layer coated probiotic microcapsule. The preparation method comprises the steps of coating probiotics before microencapsulation and coating probiotics after fermentation, thereby preparing the multilayer composite coated probiotic microcapsule. The preparation method of the invention plays a great role in maintaining the survival and stability of the probiotic preparation. In addition, the preparation method can more conveniently realize the large-scale production of the probiotic microencapsulated product, and is suitable for the preparation of the probiotic microcapsules in the fields of feed, food, medicine and the like.
Description
Technical Field
The invention belongs to the technical field of microorganisms. More particularly relates to a preparation method of a multi-layer coated probiotic microcapsule.
Background
The function of the probiotic live bacteria preparation is mainly dependent on the life activities of live bacteria contained in the probiotic live bacteria preparation, so that the content of the live bacteria, the vitality of the live bacteria and the quality guarantee period are key factors for evaluating the quality of products. At present, researchers at home and abroad maintain the survival and stability of probiotic preparations as the standard of quality evaluation. In order to solve the problems of low survival rate and instability of probiotics in use, the protection of probiotics by using a microcapsule embedding technology becomes a hot spot of domestic and foreign researches at present. However, most of the existing microcapsule coating technologies and products have the problem of porous surfaces, especially, a single-layer microcapsule has a pore size of 500-900 nm and poor stability, and most of the microcapsules cannot bear the action of low-pH-value gastric acid, bile acid, pancreatic juice and the like after entering the digestive tract of a human or an animal; when the additive is applied to livestock and poultry feed, the high-temperature and high-pressure impact of feed granulation is difficult to resist; meanwhile, the single-layer microcapsule processed still cannot meet the requirement on the shape stability of the film. In addition, some technical processes are not suitable for large-scale production, and the activity of the coated probiotics and the industrialization and application of the products thereof are seriously influenced.
Therefore, a preparation method of the microcapsule which has a great effect on maintaining the survival and stability of the probiotic preparation needs to be developed, the large-scale production of the probiotic microencapsulated product can be realized more conveniently, and the preparation method is suitable for the preparation of the probiotic microcapsule in a plurality of fields such as feed, food, medicine and the like.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the preparation method of the multilayer coated probiotic microcapsule, which has good stress resistance and good preparation stability and is suitable for industrial production.
The invention also aims to provide a multilayer coating probiotic microcapsule prepared by the method and application thereof in the fields of feed, food or medicine.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a method for preparing a multi-layer coated probiotic microcapsule, comprising:
(1) preparing a sodium alginate solution with the concentration of 10g/L-20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1-1.5 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 35-38 ℃, adding the probiotic seed liquid at the last logarithmic growth stage until the density of the probiotic is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as an oil phase, wherein the volume percentage of the span 80 is 0.1-0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1-0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400 plus 500rpm according to the volume ratio of the water phase to the oil phase of 1:3-5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500 plus 600, immobilizing for 10-20min, stopping stirring, adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10-20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) adding a culture medium suitable for growth of probiotics into a fermentation tank, feeding ammonia water at 35-38 ℃ under the condition that the stirring speed is 50-100rpm, controlling the pH value of fermentation liquor to be 6.2-6.9, continuously culturing for 8-22h until more than 80% of the space in the microcapsule is filled with the probiotics, and centrifuging to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution, and fully mixing to perform film forming reaction to obtain double-layer microcapsule bacterial sludge;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-30-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5-10 parts of glycerol, 5-10 parts of trehalose, 0.5-1 part of skimmed milk powder, 0.5-1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5-1 part of xanthan gum, 0.001-0.005 part of sodium carboxymethylcellulose, 0.2-0.8 part of microcrystalline cellulose and 100 parts of water and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) and (3) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 40-55 ℃, controlling the air outlet temperature to be 30-40 ℃, drying until the water content of the sample is 10-20%, spraying an enteric coating material to form a coating, and sieving the dried and coated sample by using a double-layer sieve to obtain a final product with the particle size of 20-60 meshes.
In the method, the probiotics is lactic acid bacteria or yeast. Wherein, the lactic acid bacteria include but not limited to one or more of lactobacillus, bifidobacterium or gram positive coccus. In a specific embodiment of the present invention, the lactic acid bacteria is one or more of lactobacillus plantarum, enterococcus faecium, enterococcus faecalis, lactobacillus reuteri, lactobacillus casei, and lactobacillus lactis.
Further, in the step (7) of the method, the chitosan acid buffer solution is prepared by preparing chitosan with molecular weight of 300-500kDa into a chitosan aqueous solution with mass concentration of 0.2-0.5%, and adjusting the pH value of the aqueous solution to 4.5-5.5.
In the step (9), the enteric coating material is 20-40% of polyacrylic resin latex solution by mass.
According to the specific embodiment of the invention, the mass ratio of the enteric coating material to the microcapsule master batch is 1:4-10, and the microcapsule master batch is sprayed at a speed of 25-50 mL/min.
In a second aspect, the invention also claims the multilayer coated probiotic microcapsule prepared by the above method and the application of the multilayer coated probiotic microcapsule in preparing feed, food or medical products.
The invention has the following beneficial effects:
the method for preparing the multi-layer coated probiotic microcapsule solves the technical problem of porous surface of the probiotic microcapsule by coating before fermentation and coating after fermentation, and greatly improves the stress resistance of the coated probiotic microcapsule and the difficult problems of easy survival after storage and feed granulation at normal temperature.
Detailed Description
In order to more clearly illustrate the invention, the invention is further described below in connection with preferred embodiments. It is to be understood by persons skilled in the art that the following detailed description is illustrative and not restrictive, and is not to be taken as limiting the scope of the invention.
The materials in the following examples, unless otherwise specified, are all materials commonly used in the art and are commercially available.
Example 1: preparation of multilayer coated enterococcus faecium microcapsule product
(1) Preparing a sodium alginate solution with the concentration of 10g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 37 ℃, adding enterococcus faecium seed liquid at the last logarithmic growth stage until the density of probiotics is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as oil phase, wherein the volume percentage of the span 80 is 0.1%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400rpm according to the volume ratio of the water phase to the oil phase of 1:3, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500, immobilizing for 10-20min, stopping stirring, then adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10, slowly settling the microcapsule, and then sucking away the oil phase;
(6) sterilizing at 121 ℃ for 30min, adding a fermentation medium cooled to 37 ℃ into a fermentation tank, adding ammonia water under the conditions of 37 ℃ and a stirring speed of 100rpm, controlling the pH value of the fermentation liquor to be 6.2-6.9, continuously culturing for 8-16h until more than 80% of the space in the microcapsule is filled with probiotics, and centrifuging at 15000r/m to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution for film forming reaction to obtain double-layer microcapsule bacterial sludge, and cleaning with sterile water; wherein the chitosan acid buffer solution is prepared by preparing chitosan with molecular weight of 400kDa into a chitosan aqueous solution with mass concentration of 0.4%, and adjusting the pH value of the aqueous solution to 5.0;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5 parts of glycerol, 10 parts of trehalose, 0.5 part of skimmed milk powder, 0.5 part of sodium acetate and 150 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5 part of xanthan gum, 0.001 part of sodium carboxymethylcellulose, 0.2 part of microcrystalline cellulose and 120 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 45 ℃, controlling the air outlet temperature to be 35 ℃, and drying for 30 min; drying until the water content of the sample is 20%, starting a side-spraying coating device, spraying enterococcus faecium microcapsule master batches by using an enteric coating material at a speed of 30mL/min (the enteric coating material is 20% by mass of polyacrylic resin emulsion, and the mass ratio of the enteric coating material to the microcapsule master batches is 1:5), forming a coating, and drying to obtain the enterococcus faecium microcapsule.
Example 2: preparation method of multilayer coated lactobacillus plantarum microcapsule product
(1) Preparing a sodium alginate solution with the concentration of 20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:5 to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 37 ℃, adding lactobacillus plantarum seed liquid at the last logarithmic growth stage until the probiotic density is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as oil phase, wherein the volume percentage of the span 80 is 0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 500rpm according to the volume ratio of the water phase to the oil phase of 1:5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:600, immobilizing for 10-20min, stopping stirring, then adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) sterilizing at 121 ℃ for 30min, adding a fermentation medium cooled to 37 ℃ into a fermentation tank, adding ammonia water under the conditions of 37 ℃ and a stirring speed of 50rpm, controlling the pH value of the fermentation liquor to be 6.2-6.9, continuously culturing for 16-22h until more than 80% of the space in the microcapsule is filled with probiotics, and centrifuging at 15000r/m to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution for film forming reaction to obtain double-layer microcapsule bacterial sludge, and cleaning with sterile water; wherein the chitosan acid buffer solution is prepared by preparing chitosan with molecular weight of 400kDa into a chitosan aqueous solution with mass concentration of 0.5%, and adjusting the pH value of the aqueous solution to 5.0;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 10 parts of glycerol, 10 parts of trehalose, 1 part of skimmed milk powder, 1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.8 part of xanthan gum, 0.003 part of sodium carboxymethyl cellulose, 0.5 part of microcrystalline cellulose and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 55 ℃, controlling the air outlet temperature to be 40 ℃, and drying for 35 min; drying until the water content of the sample is 10%, starting a side-spraying coating device, spraying enterococcus faecium microcapsule master batch by adopting an enteric coating material at the speed of 30mL/min (the enteric coating material is 30% polyacrylic resin emulsion by mass, and the mass ratio of the enteric coating material to the microcapsule master batch is 1:8), forming a coating, and drying to obtain the enterococcus faecium microcapsule.
Test example 1: simulation high-temperature granulation test result of multilayer-coated lactic acid bacteria microcapsule product
By utilizing the multilayer-coated enterococcus faecium microcapsule product and the lactobacillus plantarum microcapsule product which are provided by the method, a single-layer-coated enterococcus faecium sample ck prepared by ZL 201310218187.8 in the prior art is used as a control, a domestic pressure cooker is adopted to continuously heat 1m30s, the high-temperature and high-humidity effect generated in the feed granulation process is simulated, the change of the viable count before and after the simulated high-temperature test treatment is calculated, and the following result is obtained, wherein the average value of the high-temperature-resistant survival rate of the multilayer-coated enterococcus faecium is 68.27%, and the high-temperature-resistant survival rate of the sample is improved by 18% compared with the high-temperature-resistant survival rate of the enterococcus faecium prepared by ZL 201310218187.8 in the prior art; the high-temperature resistant survival rate of the multilayer coated lactobacillus plantarum is 53.26%, which is 32% higher than that of a lactobacillus plantarum sample prepared by using the prior art ZL 201310218187.8, and the specific experimental results are shown in Table 1.
TABLE 1 high temperature test of multilayer-coated lactic acid bacteria samples
Test example 2: test result of room temperature preservation of multilayer coated lactic acid bacteria microcapsule product
The multilayer-coated enterococcus faecium microcapsule product (example 1) and the novel-coated lactobacillus plantarum microcapsule product (example 2) prepared by the method of the invention are compared with a single-layer-coated enterococcus faecium sample and a single-layer-coated lactobacillus plantarum sample prepared by the method of the prior art ZL 201310218187.8, and non-coated bacterial powder, and are placed at 25 ℃ to observe the change of viable count of the samples in 60 days and 90 days, so that the survival rate of the samples is calculated. The result shows that the average survival rate of the multilayer-coated enterococcus faecium microcapsule product is 90% at 60 days, is 38% higher than the average survival rate of the single-layer-coated enterococcus faecium ck at 60 days by 65%, and is 80% higher than the average survival rate of the uncoated enterococcus faecium powder at 60 days by 50%; the average survival rate of the multilayer coated lactobacillus plantarum microcapsule product is 58 percent at 60 days, is 93 percent higher than the average survival rate of 30 percent at 60 days of single-layer coated lactobacillus plantarum ck, and is 480 percent higher than the average survival rate of 10 percent at 60 days of uncoated enterococcus faecium powder; the average survival rate of the multi-layer coated enterococcus faecium microcapsule product provided by the invention is 75% higher than 50% of the average survival rate of enterococcus faecium ck90 by 50% in 90 days, and is 88% higher than 40% of the average survival rate of uncoated enterococcus faecium powder by 90 days; the average survival rate of the lactobacillus plantarum microcapsule product is 49 percent at 90 days, is 390 percent higher than the average survival rate of the lactobacillus plantarum ck at 90 days by 10 percent, and is 970 percent higher than the average survival rate of the uncoated lactobacillus plantarum powder at 90 days by 10 percent.
TABLE 2 survival rates of multi-layer coated lactic acid bacteria products, pure microcapsule products and uncoated powder after storage at 25 ℃ for 60d and 90d
Test example 3: influence of different protective agent formulas on high-temperature resistance of coated lactic acid bacteria
And (3) experimental design: 4 different protective agent formulas are selected in the test, enterococcus faecium is used as an initial strain, and the coated enterococcus faecium product is prepared according to the method in the embodiment 1; meanwhile, microcapsule enterococcus faecium without protective agent is used as a control group, and the other 4 groups with protective agent are used as test groups, so as to obtain the optimal formula process parameters of the enterococcus faecium protective agent and improve the high temperature resistance of the enterococcus faecium protective agent.
Wherein, 4 experimental groups are respectively: (1) protectant 1 (same as example 1); (2) protective agent 2: adding 5 parts of glycerol and 10 parts of trehalose into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; (3) protective agent 3: adding 5 parts of glycerol, 10 parts of trehalose, 5 parts of skimmed milk powder, VC2 parts and 1 part of sodium acetate into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; (4) protective agent 4: 5 parts of glycerol, 10 parts of trehalose, 2 parts of gelatin, 2 parts of VC and 1 part of sodium acetate are added into the double-layer microcapsule bacterial sludge according to 100 parts of the weight of the double-layer microcapsule bacterial sludge.
And (3) test results: as can be seen from Table 3, the survival rate of the products produced by the group 1 protective agent after the laboratory simulation high temperature granulation treatment is significantly higher than that of the control group without the protective agent and the test groups without the protective agent.
TABLE 3 high temperature resistance test of different protectant formulations on coated lactic acid bacteria
| Group of | Before average viable count (cfu/g) | Average number of live bacteria (cfu/g) | High temperature resistant survival rate |
| Control group without protective agent | 2.7×1010 | 3.0×109 | 11.11％ |
| Protectant 1 (example 1) | 5.3×1010 | 3.8×1010 | 71.70％ |
| Protectant 2 | 2.4×1010 | 1.1×1010 | 45.83％ |
| Protectant 3 | 4.4×1010 | 1.8×1010 | 40.91％ |
| Protectant 4 | 2.8×1010 | 4.6×109 | 16.43％ |
Test example 4: study on storage stability of coated lactic acid bacteria at 25 ℃ by different protective agents
And (3) experimental design: 4 different protective agent formulas are selected in the test, enterococcus faecium is used as an initial strain, and the coated enterococcus faecium product is prepared according to the method in the embodiment 1; meanwhile, microcapsule enterococcus faecium without protective agent is used as a control group, and the other 4 groups with protective agent are used as test groups, so as to obtain the optimal formula process parameters of the enterococcus faecium protective agent and improve the storage performance of the enterococcus faecium protective agent.
Wherein, 4 experimental groups are respectively: (1) protectant 1 (same as example 1); (2) protective agent 2: adding 5 parts of glycerol and 10 parts of trehalose into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; (3) protective agent 3: adding 5 parts of glycerol, 10 parts of trehalose, 5 parts of skimmed milk powder, VC2 parts and 1 part of sodium acetate into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; (4) protective agent 4: 5 parts of glycerol, 10 parts of trehalose, 2 parts of gelatin, 2 parts of VC and 1 part of sodium acetate are added into the double-layer microcapsule bacterial sludge according to 100 parts of the weight of the double-layer microcapsule bacterial sludge.
And (3) test results: as can be seen from Table 4, the survival rates of the products produced in the protectant 1 group after being stored at 25 ℃ for 1 month, 2 months and 3 months are all obviously higher than those of the control group without the protectant and other protectant test groups.
TABLE 4 Effect of different protectant formulations on the survival rate of coated lactic acid bacteria stored at 25 deg.C for 3 months
It should be understood that the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention, and it will be obvious to those skilled in the art that other variations or modifications may be made on the basis of the above description, and all embodiments may not be exhaustive, and all obvious variations or modifications may be included within the scope of the present invention.
Claims (9)
1. A preparation method of a multi-layer coated probiotic microcapsule is characterized by comprising the following steps:
(1) preparing a sodium alginate solution with the concentration of 10g/L-20g/L in a material supplementing tank of a fermentation system;
(2) weighing calcium carbonate according to the mass ratio of 1:1-1.5 of the calcium carbonate to the sodium alginate, adding the calcium carbonate into a material supplementing tank, uniformly mixing, sterilizing, cooling to 35-38 ℃, adding the probiotic seed liquid at the last logarithmic growth stage until the density of the probiotic is 1 multiplied by 106-5×106cfu/ml, fully and uniformly mixed to be used as a water phase;
(3) adding span 80 into liquid paraffin as an oil phase, wherein the volume percentage of the span 80 is 0.1-0.3%, adding the oil phase into a fermentation tank, and sterilizing;
(4) preparing 0.1-0.3mol/L calcium chloride solution as a settling agent in another material supplementing tank, and sterilizing;
(5) slowly transferring the water phase mixed solution into the oil phase of a fermentation tank at a stirring speed of 400 plus 500rpm according to the volume ratio of the water phase to the oil phase of 1:3-5, stirring for 2-5min, then adding glacial acetic acid according to the volume ratio of the glacial acetic acid to the oil phase of 1:500 plus 600, immobilizing for 10-20min, stopping stirring, adding a settling agent into the reaction system according to the volume ratio of the glacial acetic acid to the oil phase of 1:10-20, slowly settling the microcapsule, and then sucking away the oil phase;
(6) adding a culture medium suitable for growth of probiotics into a fermentation tank, feeding ammonia water at 35-38 ℃ under the condition that the stirring speed is 50-100rpm, controlling the pH value of fermentation liquor to be 6.2-6.9, continuously culturing for 8-22h until more than 80% of the space in the microcapsule is filled with the probiotics, and centrifuging to obtain single-layer microcapsule bacterial sludge;
(7) adding the single-layer microcapsule bacterial sludge into chitosan acid buffer solution for film forming reaction to obtain double-layer microcapsule bacterial sludge;
(8) adding a protective agent into the double-layer microcapsule bacterial sludge obtained in the step (7), uniformly mixing, adding a packaging material solution, and preparing 20-30-mesh microcapsule master batches in a granulator; wherein, the protective agent comprises the following components: adding 5-10 parts of glycerol, 5-10 parts of trehalose, 0.5-1 part of skimmed milk powder, 0.5-1 part of sodium acetate and 300 parts of corn starch into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts; the packaging material solution comprises the following components: adding 0.5-1 part of xanthan gum, 0.001-0.005 part of sodium carboxymethylcellulose, 0.2-0.8 part of microcrystalline cellulose and 100 parts of water and 200 parts of water into the double-layer microcapsule bacterial sludge by taking the weight of the double-layer microcapsule bacterial sludge as 100 parts;
(9) and (3) transferring the microcapsule master batch prepared in the step (8) into a fluidized granulation coating dryer, controlling the air inlet temperature to be 40-55 ℃, controlling the air outlet temperature to be 30-40 ℃, drying until the water content of the sample is 10-20%, spraying an enteric coating material to form a coating, and sieving the dried and coated sample by using a double-layer sieve to obtain the multilayer coated probiotic microcapsule with the particle size of 20-60 meshes.
2. The method of claim 1, wherein the probiotic bacteria are lactic acid bacteria or yeast.
3. The method according to claim 2, wherein the lactic acid bacteria is one or more of lactobacillus, bifidobacterium or gram-positive coccus.
4. The method according to claim 2, wherein the lactic acid bacteria is one or more of Lactobacillus plantarum, enterococcus faecium, enterococcus faecalis, Lactobacillus reuteri, Lactobacillus casei, and Lactobacillus lactis.
5. The preparation method according to claim 1, wherein in the step (7), the chitosan acidic buffer solution is prepared by preparing chitosan with molecular weight of 300-500kDa into 0.2-0.5% by mass of chitosan aqueous solution, and adjusting the pH value of the aqueous solution to 4.5-5.5.
6. The preparation method according to claim 1, wherein in the step (9), the enteric coating material is 20-40% by mass of polyacrylic resin latex.
7. The preparation method according to claim 1, wherein in the step (9), the mass ratio of the enteric coating material to the microcapsule masterbatch is 1:4-10, and the microcapsule masterbatch is sprayed at a speed of 25-50 mL/min.
8. Multilayer coated probiotic microcapsules prepared according to the preparation process of any one of claims 1 to 7.
9. Use of the multi-layer coated probiotic microcapsule according to claim 8 for the preparation of feed, food or pharmaceutical products.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103222539A (en) * | 2013-04-09 | 2013-07-31 | 魏永刚 | Preparation method of microbial pre-fermentation coating multilayer microcapsule |
| CN103275966A (en) * | 2013-06-04 | 2013-09-04 | 国家粮食局科学研究院 | Method for preparing coating microcapsule before microbial fermentation by utilizing common fermentation tank |
| CN103275962A (en) * | 2013-06-04 | 2013-09-04 | 国家粮食局科学研究院 | Method for preparing coated microcapsules before microbial fermentation |
| CN105534951A (en) * | 2016-01-08 | 2016-05-04 | 魏永刚 | Synbiotic microcapsules and preparing method and application thereof |
| CN105567669A (en) * | 2016-01-08 | 2016-05-11 | 魏永刚 | Probiotics microcapsule preparation and preparation method thereof |
| CN109170235A (en) * | 2018-08-01 | 2019-01-11 | 中国农业大学 | Probiotic microcapsule and the preparation method and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101355003B1 (en) * | 2010-11-02 | 2014-01-24 | 정명준 | Lactic acid bacteria having multi coating layers and preparing method thereof |
| ITMI20120131A1 (en) * | 2012-02-01 | 2013-08-02 | Probiotical Spa | MULTILAYER MICROCAPSULATED PROBIOTIC BACTERIA, THEIR PRODUCTION AND USE |
| TR201712406A2 (en) * | 2017-08-21 | 2019-03-21 | Nanomik Biyoteknoloji A S | Probiotic Loaded Microcapsules and Their Production |
-
2020
- 2020-11-03 CN CN202011208280.7A patent/CN112544977B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN103222539A (en) * | 2013-04-09 | 2013-07-31 | 魏永刚 | Preparation method of microbial pre-fermentation coating multilayer microcapsule |
| CN103275966A (en) * | 2013-06-04 | 2013-09-04 | 国家粮食局科学研究院 | Method for preparing coating microcapsule before microbial fermentation by utilizing common fermentation tank |
| CN103275962A (en) * | 2013-06-04 | 2013-09-04 | 国家粮食局科学研究院 | Method for preparing coated microcapsules before microbial fermentation |
| CN105534951A (en) * | 2016-01-08 | 2016-05-04 | 魏永刚 | Synbiotic microcapsules and preparing method and application thereof |
| CN105567669A (en) * | 2016-01-08 | 2016-05-11 | 魏永刚 | Probiotics microcapsule preparation and preparation method thereof |
| CN109170235A (en) * | 2018-08-01 | 2019-01-11 | 中国农业大学 | Probiotic microcapsule and the preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| "发酵前及发酵后包被粪肠球菌";张琳 等;《粮油食品科技》;20161231;第24卷(第2期);80-84 * |
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