CN111419822A - Calcium butyrate microcapsule and preparation method and application thereof - Google Patents
Calcium butyrate microcapsule and preparation method and application thereof Download PDFInfo
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- CN111419822A CN111419822A CN202010338469.1A CN202010338469A CN111419822A CN 111419822 A CN111419822 A CN 111419822A CN 202010338469 A CN202010338469 A CN 202010338469A CN 111419822 A CN111419822 A CN 111419822A
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- Prior art keywords
- calcium butyrate
- butyrate
- calcium
- microcapsule
- preparation
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- FYPVXEILSNEKOO-UHFFFAOYSA-L calcium;butanoate Chemical compound [Ca+2].CCCC([O-])=O.CCCC([O-])=O FYPVXEILSNEKOO-UHFFFAOYSA-L 0.000 title claims abstract description 60
- 239000003094 microcapsule Substances 0.000 title claims abstract description 46
- 238000002360 preparation method Methods 0.000 title claims abstract description 24
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 claims abstract description 49
- 241000193171 Clostridium butyricum Species 0.000 claims abstract description 40
- 239000000463 material Substances 0.000 claims abstract description 27
- 230000001580 bacterial effect Effects 0.000 claims abstract description 25
- 238000000855 fermentation Methods 0.000 claims abstract description 21
- 239000003814 drug Substances 0.000 claims abstract description 15
- 239000002775 capsule Substances 0.000 claims abstract description 14
- 230000004151 fermentation Effects 0.000 claims abstract description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 42
- 108010067770 Endopeptidase K Proteins 0.000 claims description 24
- 239000011780 sodium chloride Substances 0.000 claims description 21
- 239000010802 sludge Substances 0.000 claims description 19
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 16
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 16
- 239000001963 growth medium Substances 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 15
- 239000000843 powder Substances 0.000 claims description 13
- 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 claims description 12
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- 239000008103 glucose Substances 0.000 claims description 12
- 230000003321 amplification Effects 0.000 claims description 11
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 11
- 241001052560 Thallis Species 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 239000011575 calcium Substances 0.000 claims description 9
- 239000000839 emulsion Substances 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 239000000725 suspension Substances 0.000 claims description 9
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 8
- 206010011732 Cyst Diseases 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 8
- 235000019270 ammonium chloride Nutrition 0.000 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 238000004140 cleaning Methods 0.000 claims description 8
- 235000005822 corn Nutrition 0.000 claims description 8
- 208000031513 cyst Diseases 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 8
- 229940099596 manganese sulfate Drugs 0.000 claims description 8
- 239000011702 manganese sulphate Substances 0.000 claims description 8
- 235000007079 manganese sulphate Nutrition 0.000 claims description 8
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 8
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 8
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- 229920002472 Starch Polymers 0.000 claims description 7
- 239000003674 animal food additive Substances 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 7
- 239000002244 precipitate Substances 0.000 claims description 7
- 238000011218 seed culture Methods 0.000 claims description 7
- 238000001694 spray drying Methods 0.000 claims description 7
- 235000019698 starch Nutrition 0.000 claims description 7
- 239000002518 antifoaming agent Substances 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000008107 starch Substances 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 239000002609 medium Substances 0.000 claims description 5
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 4
- 244000105624 Arachis hypogaea Species 0.000 claims description 4
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 4
- 235000018262 Arachis monticola Nutrition 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 4
- 235000010469 Glycine max Nutrition 0.000 claims description 4
- 239000001888 Peptone Substances 0.000 claims description 4
- 108010080698 Peptones Proteins 0.000 claims description 4
- 235000015278 beef Nutrition 0.000 claims description 4
- 238000006243 chemical reaction Methods 0.000 claims description 4
- 235000020232 peanut Nutrition 0.000 claims description 4
- 235000019319 peptone Nutrition 0.000 claims description 4
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- 238000010008 shearing Methods 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 abstract description 8
- 230000000873 masking effect Effects 0.000 abstract description 6
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 abstract description 5
- 241001465754 Metazoa Species 0.000 abstract description 5
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- MFBOGIVSZKQAPD-UHFFFAOYSA-M sodium butyrate Chemical compound [Na+].CCCC([O-])=O MFBOGIVSZKQAPD-UHFFFAOYSA-M 0.000 abstract description 3
- 239000002904 solvent Substances 0.000 abstract description 3
- 231100000331 toxic Toxicity 0.000 abstract description 3
- 230000002588 toxic effect Effects 0.000 abstract description 3
- 150000001720 carbohydrates Chemical class 0.000 abstract description 2
- 239000004615 ingredient Substances 0.000 abstract description 2
- 235000003170 nutritional factors Nutrition 0.000 abstract description 2
- 235000013343 vitamin Nutrition 0.000 abstract description 2
- 239000011782 vitamin Substances 0.000 abstract description 2
- 229940088594 vitamin Drugs 0.000 abstract description 2
- 229930003231 vitamin Natural products 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 5
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- 239000012528 membrane Substances 0.000 description 5
- 239000000919 ceramic Substances 0.000 description 4
- 238000006386 neutralization reaction Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 108091005804 Peptidases Proteins 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000005034 decoration Methods 0.000 description 2
- 238000005189 flocculation Methods 0.000 description 2
- 230000016615 flocculation Effects 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000009965 odorless effect Effects 0.000 description 2
- 235000019419 proteases Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000009777 vacuum freeze-drying Methods 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 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 description 1
- RSWGJHLUYNHPMX-UHFFFAOYSA-N Abietic-Saeure Natural products C12CCC(C(C)C)=CC2=CCC2C1(C)CCCC2(C)C(O)=O RSWGJHLUYNHPMX-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 241000193403 Clostridium Species 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- 239000001856 Ethyl cellulose Substances 0.000 description 1
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 239000005062 Polybutadiene Substances 0.000 description 1
- 239000004698 Polyethylene Substances 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- KHPCPRHQVVSZAH-HUOMCSJISA-N Rosin Natural products O(C/C=C/c1ccccc1)[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 KHPCPRHQVVSZAH-HUOMCSJISA-N 0.000 description 1
- 229920001800 Shellac Polymers 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000001785 acacia senegal l. willd gum Substances 0.000 description 1
- 239000002535 acidifier Substances 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001768 carboxy methyl cellulose Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
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- 239000000110 cooling liquid Substances 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 235000019325 ethyl cellulose Nutrition 0.000 description 1
- 229920001249 ethyl cellulose Polymers 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 230000003311 flocculating effect Effects 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000009655 industrial fermentation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003871 intestinal function Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 229920002857 polybutadiene Polymers 0.000 description 1
- -1 polyethylene Polymers 0.000 description 1
- 229920000573 polyethylene Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 238000011085 pressure filtration Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000013341 scale-up Methods 0.000 description 1
- 239000004208 shellac Substances 0.000 description 1
- 235000013874 shellac Nutrition 0.000 description 1
- 229940113147 shellac Drugs 0.000 description 1
- ZLGIYFNHBLSMPS-ATJNOEHPSA-N shellac Chemical compound OCCCCCC(O)C(O)CCCCCCCC(O)=O.C1C23[C@H](C(O)=O)CCC2[C@](C)(CO)[C@@H]1C(C(O)=O)=C[C@@H]3O ZLGIYFNHBLSMPS-ATJNOEHPSA-N 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
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- 238000001179 sorption measurement Methods 0.000 description 1
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- 239000000126 substance Substances 0.000 description 1
- KHPCPRHQVVSZAH-UHFFFAOYSA-N trans-cinnamyl beta-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OCC=CC1=CC=CC=C1 KHPCPRHQVVSZAH-UHFFFAOYSA-N 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- 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
- A61K9/5063—Compounds of unknown constitution, e.g. material from plants or animals
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Physiology (AREA)
- Botany (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a calcium butyrate microcapsule and a preparation method and application thereof, relating to the technical field of microcapsules; the thallus in the fermentation process of the sodium butyrate is used as a capsule material, after the fermentation of clostridium butyricum is finished, the thallus is broken, so that butyric acid is adsorbed into cells again, and the odor of the butyrate is prevented from volatilizing by re-embedding the butyric acid by the thallus. The calcium butyrate microcapsule disclosed by the invention is simple in preparation process, no additional chemical reagent is added, no toxic or harmful chemical reagent is basically required to be introduced, and no danger of solvent residue exists; the capsule material using the thallus per se has better taste masking effect, and can realize the slow release of the medicine to a certain extent; mycoprotein, saccharides, vitamins, nutritional factors and the like in the bacterial cells can be used as nutritional ingredients for animals.
Description
Technical Field
The invention belongs to the technical field of microcapsules, and particularly relates to a calcium butyrate microcapsule as well as a preparation method and application thereof.
Background
Clostridium butyricum was first discovered and reported in 1933 by the university of qianye medical university of japan, womb recently doctor, and is therefore also known as a species of introgression, classified as a clostridium species, a gram-positive anaerobic bacillus. In 1935, Rousi institute of microbiology, doctor Kingimiyairi, isolated Clostridium butyricum from human feces and soil, and subsequently discovered that the filtrate of anaerobic culture contained less fatty acid and had a strong intestinal function, it could inhibit pathogenic bacteria in intestinal tract and promote the growth of beneficial bacteria such as bifidobacteria and lactobacilli in intestinal tract, and introduced Clostridium butyricum in the research institute of Russia, institute of microbiology, department of sciences of Heilongjiang province, 1992, and colonized in China, made a significant contribution to the development of beneficial bacteria in China.
The main metabolite of clostridium butyricum is butyric acid, and a butyrate product can be obtained through industrial fermentation culture of clostridium butyricum. Butyrate is one of the feed additive varieties allowed to be used in China, and is listed in feed additive variety catalog by the Ministry of agriculture in 2006. The application mode in animal production is mainly characterized in that butyrate products are added into animal feed to play roles in intestinal mucosa nutritional agents, electrolyte balance regulators, gastrointestinal microecological regulators, compound acidifier, flavoring agents, feeding promoting agents and the like.
Normally, sodium butyrate has a strong lipid odor. In the production process of the feed, strong fat odor causes great interference to the production industry and the production environment, and after the feed is added, a feed factory still emits fat odor in the processing process, and both manufacturers and users are subjected to strong environmental protection pressure.
The microcapsule technology is a preparation product with millimeter to micron level formed by embedding medicines by using natural high molecular materials (gelatin, Arabic gum, shellac, starch, dextrin, wax, rosin, sodium alginate, chitosan and the like) or semi/total synthetic high molecular materials (carboxymethyl cellulose, methyl cellulose, ethyl cellulose, polyethylene, polystyrene, polybutadiene and the like) as capsule materials. The drug is embedded by the microcapsule technology to cover the unpleasant smell of the drug, but the traditional microcapsule technology needs to additionally use a plurality of chemical reagents, so that the chemical reagents of the product are remained, the process is complex, and the taste-covering effect is not good.
Disclosure of Invention
In view of the above, the invention aims to provide a calcium butyrate microcapsule, a preparation method and an application thereof, which have the advantages of simple process, no need of adding extra chemical reagents and good taste masking effect.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a preparation method of calcium butyrate microcapsules, which comprises the following steps: (1) inoculating clostridium butyricum into a seed culture medium for culture, collecting thalli in a culture solution, and cleaning to obtain bacterial sludge;
(2) adding a NaCl solution and proteinase K into the bacterial sludge, stirring for 1-3 h at 55-60 ℃, performing wall breaking treatment, collecting bacterial precipitates after wall breaking, and performing freeze drying to obtain a clostridium butyricum cell cyst material;
(3) reacting butyric acid with Ca (OH)2Reacting to generate a calcium butyrate solution, and mixing the calcium butyrate solution with protease K to break the wall to obtain a wall-broken solution;
(4) mixing the wall breaking liquid with the clostridium butyricum cell cyst material obtained in the step (2), stirring, and drying to obtain calcium butyrate microcapsules;
no chronological sequence exists among the steps (1), (2) and (3).
Preferably, the culturing in the step (1) comprises two-stage amplification culturing, wherein a culture medium of the first-stage amplification culturing comprises the following components in percentage by mass: 2% of glucose, 1% of yeast extract, 1% of tuna extract, 1% of peptone, 0.5% of soluble starch, 0.5% of sodium chloride, 0.05% of glycerol and the balance of water;
the culture medium for the second-stage amplification culture comprises the following components in percentage by mass: 2.5% of glucose, 2.0% of corn steep liquor, 3.5% of yeast extract, 1% of beef extract, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of monopotassium phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate and the balance of water.
Preferably, the volume ratio of the bacterial sludge to the NaCl solution in the step (2) is 1: 20-30; the mass percentage of NaCl in the NaCl solution is 3-5%;
the volume ratio of the bacterial sludge to the proteinase K is 1:10-4～10-3The enzyme activity of the proteinase K is 40U/mg.
Preferably, step (3) Ca (OH)2Is suspension emulsion after high-speed shearing; ca (OH) in the suspension emulsion2The mass percentage content of (A) is 20%;
the volume ratio of butyric acid to the suspension emulsion during the reaction in the step (3) is 1: 10-30;
the volume ratio of the calcium butyrate solution to the proteinase K is 1:10-3～10-2。
Preferably, the butyric acid comprises butyric acid obtained by anaerobic fermentation of clostridium butyricum; the fermentation medium used for anaerobic fermentation comprises the following components in percentage by mass: 3% of glucose, 2.5% of corn steep liquor dry powder, 3.5% of peanut cake powder, 1.5% of soybean cake powder, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of potassium dihydrogen phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate, 0.05% of defoaming agent and the balance of water
Preferably, the stirring time in the step (4) is 2-3 h.
Preferably, the drying in the step (4) comprises spray drying, wherein the inlet air temperature of the tower is 170-270 ℃ and the outlet air temperature of the tower is 70-170 ℃ during the spray drying.
The invention also provides a calcium butyrate microcapsule, wherein the capsule material of the calcium butyrate microcapsule is the thallus of clostridium butyricum, and the mass ratio of the capsule material to calcium butyrate is (5-60): 40 to 95.
The invention also provides application of the calcium butyrate microcapsule in preparation of a taste-masking feed additive.
The invention also provides application of the calcium butyrate microcapsule in preparation of a slow-release medicament.
The invention provides a preparation method of calcium butyrate microcapsules, which is characterized in that after fermentation of clostridium butyricum, butyric acid is adsorbed into cells again through wall breaking treatment of thalli, and odor volatilization of butyrate is avoided through re-embedding of the thalli on the butyric acid. The invention takes the thalli in the sodium butyrate fermentation process as the capsule wall material, is not a traditional high molecular material, has simple process, does not need to add extra chemical reagents, basically does not need to introduce toxic and harmful chemical reagents, and has no danger of solvent residue; the capsule material using the thallus per se has better taste masking effect, and can realize the slow release of the medicine to a certain extent; mycoprotein, saccharides, vitamins, nutritional factors and the like in the bacterial cells can be used as nutritional ingredients for animals.
The clostridium butyricum is a natural material, compared with a traditional synthesized water-soluble material, the clostridium butyricum has stronger flexibility, toughness and stability, the prepared clostridium butyricum microcapsule not only keeps the characteristics of the function and the like of the medicine, but also has rich nutrient substances of the clostridium butyricum, and the clostridium butyricum microcapsule of the calcium butyrate has no toxicity, high biocompatibility and excellent dispersibility in water, can realize the slow release of the medicine and has stable biological activity of the medicine; the size of the microcapsule is dozens of microns, and the microcapsule is freeze-dried into powder, so that the microcapsule is convenient to apply and easy to store; the preparation condition is mild, the preparation equipment is simple, the operation is convenient, toxic and harmful chemical reagents do not need to be introduced basically, the danger of solvent residue does not exist, and the method is very suitable for embedding medicaments and food additives.
Detailed Description
The invention provides a preparation method of calcium butyrate microcapsules, which comprises the following steps: (1) inoculating clostridium butyricum into a seed culture medium for culture, collecting thalli in a culture solution, and cleaning to obtain bacterial sludge;
(2) adding a NaCl solution and proteinase K into the bacterial sludge, stirring for 1-3 h at 55-60 ℃, performing wall breaking treatment, collecting bacterial precipitates after wall breaking, and performing freeze drying to obtain a clostridium butyricum cell cyst material;
(3) reacting butyric acid with Ca (OH)2Reacting to generate a calcium butyrate solution, and mixing the calcium butyrate solution with protease K to break the wall to obtain a wall-broken solution;
(4) mixing and stirring the wall breaking liquid and the clostridium butyricum cell cyst material obtained in the step (2), and drying to obtain calcium butyrate microcapsules;
no chronological sequence exists among the steps (1), (2) and (3).
The method comprises the steps of inoculating clostridium butyricum into a seed culture medium for culture, collecting thalli in a culture solution, and cleaning to obtain bacterial sludge. The culture of the invention preferably comprises two-stage amplification culture, wherein the culture medium of the first-stage amplification culture preferably comprises the following components in percentage by mass: 2% of glucose, 1% of yeast extract, 1% of tuna extract, 1% of peptone, 0.5% of soluble starch, 0.5% of sodium chloride, 0.05% of glycerol and the balance of water; wherein the culture medium for the second-stage amplification culture preferably comprises the following components in percentage by mass: 2.5% of glucose, 2.0% of corn steep liquor, 3.5% of yeast extract, 1% of beef extract, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of monopotassium phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate and the balance of water. The source of the soluble starch is not particularly limited in the present invention, and conventional soluble starches in the art may be used. The present invention is not particularly limited to the inoculation and scale-up culture protocol, and may be a routine protocol in the art. The thalli are obtained by preferably centrifuging, flocculating or membrane filtering the culture solution, wherein the centrifugation is preferably carried out at the rotating speed of 6000rpm for 15min, the flocculation preferably comprises the flocculation by using 0.15 percent by mass of polyacrylamide, and the membrane filtration preferably comprises the low-pressure (0.1MP) filtration by using a 50nm ceramic membrane. The bacteria are cleaned to obtain the bacterial sludge, the cleaning method preferably comprises ceramic membrane low-pressure filtration cleaning, the amount of cleaning water is added according to actual filtration conditions, and the ceramic membrane preferably comprises a 50nm ceramic membrane.
After bacterial sludge is obtained, adding a NaCl solution and proteinase K into the bacterial sludge, stirring for 1-3 h at 55-60 ℃, performing wall breaking treatment, collecting bacterial precipitates after wall breaking, and freeze-drying to obtain the clostridium butyricum cell cyst material. When NaCl solution and proteinase K are added into the bacterial sludge, the NaCl solution is preferably added firstly, then the pH value is adjusted to 6.0, and finally the proteinase K is added. The volume ratio of the bacterial sludge to the NaCl solution is preferably 1:20 to 30. The mass percentage of NaCl in the NaCl solution is preferably 3-5%. The volume ratio of the bacterial sludge to the proteinase K is preferably 1:10-4～10-3The method comprises the steps of cooling liquid subjected to wall breaking treatment to room temperature, centrifuging and washing for 3 times, and collecting thalli precipitates, wherein the liquid is preferably subjected to wall breaking treatment in a 50L glass reaction kettle, wherein the wall breaking treatment is preferably carried out at 37 ℃ and 200rpm, and stirring for 60min at normal pressure, the centrifugal rotation speed is preferably 6000r/min, the time is preferably 15min, the thalli precipitates subjected to wall breaking are collected and freeze-dried, the freeze-drying is preferably vacuum freeze-drying, the vacuum freeze-drying temperature is preferably-50 ℃, the vacuum degree is preferably-0.1 MP, and the drying time is preferably 1.5 h.
The invention relates to butyric acid and Ca (OH)2And mixing the solutions, performing neutralization reaction to generate a calcium butyrate solution, and mixing the calcium butyrate solution with protease K to break the wall to obtain a wall-broken solution. The butyric acid is preferably butyric acid obtained by anaerobic fermentation of clostridium butyricum, the method for producing butyric acid by anaerobic fermentation of clostridium butyricum is not particularly limited, and preferably comprises inoculating clostridium butyricum into the seed culture medium for culture to obtain seed solution, performing anaerobic fermentation on the seed solution,thus obtaining the butyric acid. The culturing method of the present invention is preferably the same as that in step (1), and will not be described herein. According to the invention, the seed solution is preferably inoculated into a fermentation medium for anaerobic fermentation, and the fermentation medium preferably comprises the following components in percentage by mass: 3% of glucose, 2.5% of corn steep liquor dry powder, 3.5% of peanut cake powder, 1.5% of soybean cake powder, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of potassium dihydrogen phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate, 0.05% of defoaming agent and the balance of water. The time of the anaerobic fermentation is preferably 45 hours, and the temperature is preferably 37 ℃. The kind and source of the defoaming agent are not particularly limited in the present invention, and it is preferably a conventional commercially available defoaming agent in the art, and a defoamer is used in the examples of the present invention.
The invention utilizes butyric acid and Ca (OH)2A neutralization reaction occurs to produce calcium butyrate, the Ca (OH)2Preferably suspension emulsion after high-speed shearing; ca (OH) in the suspension emulsion2The mass percentage of (B) is 20%. In the neutralization reaction of the present invention, the volume ratio of the butyric acid to the suspension emulsion is preferably 1:10 to 30. After the calcium butyrate solution is obtained, the calcium butyrate solution is mixed with protease K and then wall breaking is carried out, and the volume ratio of the calcium butyrate solution to the protease K is preferably 1:10-3～10-2The enzyme activity of the proteinase K is preferably 40U/mg. The wall breaking method is preferably carried out for 1-3 h after mixing and stirring, and the stirring temperature is preferably 55-60 ℃.
After the wall-broken liquid is obtained, the wall-broken liquid and the clostridium butyricum cell capsule wall material obtained in the step (2) are mixed, stirred and dried to obtain the calcium butyrate microcapsule. After the mixing, the pH value is preferably adjusted to 5.5-8.0, calcium butyrate in the wall-broken liquid is adsorbed by using the clostridium butyricum cell envelope material, and the stirring is carried out for 2-3 hours while the adsorption is carried out. The drying is preferably spray drying, the tower inlet air temperature is preferably 170-270 ℃ and the outlet air temperature is preferably 70-170 ℃ during spray drying.
The invention provides a calcium butyrate microcapsule, wherein a capsule material of the calcium butyrate microcapsule is a thallus of clostridium butyricum, and the mass ratio of the capsule material to calcium butyrate is (5-60): 40 to 95. In the calcium butyrate microcapsule, the mass ratio of the capsule material to the calcium butyrate is preferably 45-55: 65-75. The particle size of the calcium butyrate microcapsule is preferably 6 microns.
The invention also provides application of the calcium butyrate microcapsule in preparation of a taste-masking feed additive. The invention has better taste masking effect by using the thallus as the capsule material. When the calcium butyrate microcapsules are used as the taste-masking feed additive, different amounts of the taste-masking feed additive are preferably added at different stages according to different animal types, and more preferably 500-1000 g/ton.
The invention also provides application of the calcium butyrate microcapsule in preparation of a slow-release medicament. The calcium butyrate microcapsule disclosed by the invention can realize slow release of a medicament to a certain extent, and can be applied to preparation of a slow-release medicament. When the calcium butyrate microcapsule is used for preparing a slow-release medicament, the calcium butyrate microcapsule is used as a slow-release framework material, and the addition amount is preferably 5-10% (mass percentage content).
The calcium butyrate microcapsules provided by the present invention, the preparation method and the application thereof are described in detail below with reference to the examples, but they should not be construed as limiting the scope of the present invention.
Example 1
Step one, inoculating clostridium butyricum into a seed culture medium for step-by-step amplification culture, centrifuging the obtained culture solution, and cleaning to obtain bacterial sludge;
wherein, the primary culture medium comprises the following components in percentage by weight: 2% of glucose, 1% of yeast extract, 1% of tuna extract, 1% of peptone, 0.5% of soluble starch, 0.5% of sodium chloride, 0.05% of glycerol and the balance of water. The secondary culture medium comprises the following components in percentage by weight: 2.5% of glucose, 2.0% of corn steep liquor, 3.5% of yeast extract, 1% of beef extract, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of monopotassium phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate and the balance of water.
Secondly, adding NaCl solution into the prepared bacterial sludge, adjusting the pH value to 6.0, simultaneously adding proteinase K, and stirring for 3 hours at 55 ℃ to perform wall breaking treatment; cooling the wall-broken bacteria to room temperature, centrifuging, washing with water for 3 times, collecting bacteria precipitate, and freeze-drying to obtain Clostridium butyricum cell cyst material for later use;
wherein, the concentration of NaCl solution is 5%, the adding proportion is 1: 24; the adding ratio of the protease K is 1:10-3(V/V), the enzyme activity of the protease is 40U/mg;
thirdly, inoculating clostridium butyricum into a seed culture medium for step-by-step amplification culture to prepare a seed solution;
fourthly, inoculating the seed liquid into a fermentation culture medium for anaerobic fermentation to generate butyric acid;
wherein the fermentation medium comprises the following components in percentage by weight: 3% of glucose, 2.5% of corn steep liquor dry powder, 3.5% of peanut cake powder, 1.5% of soybean cake powder, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of potassium dihydrogen phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate, 0.05% of defoaming agent and the balance of water.
The fifth step, add 20% Ca (OH) to the fermentation product at a ratio of 1:202Carrying out neutralization reaction to generate calcium butyrate;
and sixthly, adding protease K into the calcium butyrate obtained in the fifth step, stirring for 3 hours at 55 ℃ to perform clostridium butyricum wall breaking treatment, adding the clostridium butyricum cell envelope prepared in the step 2 after wall breaking is finished, adjusting the pH to 7.0, adsorbing the calcium butyrate in the wall breaking liquid by using the clostridium butyricum cell envelope, and stirring for 3 hours while adsorbing.
Wherein the adding ratio of the protease K is 1:10-2(V/V), the enzyme activity of the protease is 40U/mg;
and seventhly, carrying out spray drying on the solution obtained in the sixth step to obtain the microcapsule containing 35% of clostridium butyricum fermented and 65% of calcium butyrate.
Wherein the air inlet temperature of the spray drying tower is 180-220 ℃, and the air outlet temperature is 70-90 ℃.
And (3) carrying out taste masking effect measurement on the prepared calcium butyrate microcapsules: and (3) respectively placing the equivalent product (sample A) and the commercially available common calcium butyrate product (sample B) in a closed container (the sample loading is not less than 2/3 of the container), and preparing 10 samples to be detected according to the sample loading. Selecting 10 healthy volunteers, randomly selecting one sample to be tested for each volunteer, testing the test feeling of each volunteer on the product, and evaluating the taste masking effect by taking severe odor, stink odor, slight odor and odorless as evaluation indexes. The product of the invention is mainly characterized by slight odor (40%) and odorless (60%), and the common products sold in the market are marked by severe odor (60%), odor (30%) and slight odor (10%).
The release degree of the prepared calcium butyrate microcapsules is measured according to a first method XC in the appendix of the second part of the pharmacopoeia of the people's republic of China, edition 2005. The release amount of the artificial gastric juice in 2 hours of the product prepared by the invention is about 5.0 percent; the release amount in 2h in the artificial intestinal juice is about 30 percent, the release amount in 12h reaches 86 percent, the release amount in 4h in the artificial intestinal juice is about 40 to 45 percent, the enteric slow release effect is achieved, and the product prepared under the condition with good in vitro release characteristic can meet the quality requirements of Chinese pharmacopoeia on slow release preparations.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (10)
1. The preparation method of the calcium butyrate microcapsule is characterized by comprising the following steps of: (1) inoculating clostridium butyricum into a seed culture medium for culture, collecting thalli in a culture solution, and cleaning to obtain bacterial sludge;
(2) adding a NaCl solution and proteinase K into the bacterial sludge, stirring for 1-3 h at 55-60 ℃, performing wall breaking treatment, collecting bacterial precipitates after wall breaking, and performing freeze drying to obtain a clostridium butyricum cell cyst material;
(3) reacting butyric acid with Ca (OH)2Generating a calcium butyrate solution after reaction, mixing the calcium butyrate solution with protease K, and breaking the wall to obtain a wall-broken solution;
(4) mixing the wall breaking liquid with the clostridium butyricum cell cyst material obtained in the step (2), stirring, and drying to obtain calcium butyrate microcapsules;
no chronological sequence exists among the steps (1), (2) and (3).
2. The method according to claim 1, wherein the culturing in step (1) comprises two-stage amplification culture, wherein the culture medium of the one-stage amplification culture comprises the following components in percentage by mass: 2% of glucose, 1% of yeast extract, 1% of tuna extract, 1% of peptone, 0.5% of soluble starch, 0.5% of sodium chloride, 0.05% of glycerol and the balance of water;
the culture medium for the second-stage amplification culture comprises the following components in percentage by mass: 2.5% of glucose, 2.0% of corn steep liquor, 3.5% of yeast extract, 1% of beef extract, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of monopotassium phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate and the balance of water.
3. The method according to claim 1, wherein the volume ratio of the bacterial sludge to the NaCl solution in the step (2) is 1: 20-30; the mass percentage of NaCl in the NaCl solution is 3-5%;
the volume ratio of the bacterial sludge to the proteinase K is 1:10-4～10-3The enzyme activity of the proteinase K is 40U/mg.
4. The method according to claim 1, wherein the Ca (OH) in the step (3)2Is suspension emulsion after high-speed shearing; ca (OH) in the suspension emulsion2The mass percentage content of (A) is 20%;
the volume ratio of butyric acid to the suspension emulsion during the reaction in the step (3) is 1: 10-30;
the volume ratio of the calcium butyrate solution to the proteinase K is 1:10-3～10-2。
5. The method according to claim 4, wherein the butyric acid comprises butyric acid obtained by anaerobic fermentation using Clostridium butyricum; the fermentation medium used for anaerobic fermentation comprises the following components in percentage by mass: 3% of glucose, 2.5% of corn steep liquor dry powder, 3.5% of peanut cake powder, 1.5% of soybean cake powder, 0.2% of ammonium sulfate, 0.5% of mycoprotein, 0.2% of potassium dihydrogen phosphate, 0.2% of calcium chloride, 0.1% of ammonium chloride, 0.07% of magnesium sulfate, 0.02% of manganese sulfate, 0.05% of defoaming agent and the balance of water.
6. The preparation method according to claim 1, wherein the stirring time in the step (4) is 2-3 h.
7. The preparation method according to claim 1, wherein the drying in the step (4) comprises spray drying, and the tower inlet air temperature is 170-270 ℃ and the outlet air temperature is 70-170 ℃.
8. The calcium butyrate microcapsule is characterized in that a capsule material of the calcium butyrate microcapsule is a thallus of clostridium butyricum, and the mass ratio of the capsule material to calcium butyrate is (5-60): 40 to 95.
9. Use of calcium butyrate microcapsules according to claim 8 for the preparation of a taste-masked feed additive.
10. Use of calcium butyrate microcapsules according to claim 8 for the preparation of a slow-release medicament.
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