CN117122064A - Liquid microcapsule system and preparation method and application thereof - Google Patents
Liquid microcapsule system and preparation method and application thereof Download PDFInfo
- Publication number
- CN117122064A CN117122064A CN202311158677.3A CN202311158677A CN117122064A CN 117122064 A CN117122064 A CN 117122064A CN 202311158677 A CN202311158677 A CN 202311158677A CN 117122064 A CN117122064 A CN 117122064A
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- China
- Prior art keywords
- parts
- liquid
- liquid microcapsule
- microcapsule system
- microcapsule
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- 239000003094 microcapsule Substances 0.000 title claims abstract description 154
- 239000007788 liquid Substances 0.000 title claims abstract description 147
- 238000002360 preparation method Methods 0.000 title claims abstract description 34
- 150000003904 phospholipids Chemical class 0.000 claims abstract description 53
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 49
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 48
- 239000005913 Maltodextrin Substances 0.000 claims abstract description 46
- 230000001580 bacterial effect Effects 0.000 claims abstract description 46
- 229920002774 Maltodextrin Polymers 0.000 claims abstract description 44
- 229940035034 maltodextrin Drugs 0.000 claims abstract description 44
- 239000000839 emulsion Substances 0.000 claims abstract description 38
- 238000000034 method Methods 0.000 claims abstract description 34
- 108010073771 Soybean Proteins Proteins 0.000 claims abstract description 30
- 230000000813 microbial effect Effects 0.000 claims abstract description 28
- 238000012545 processing Methods 0.000 claims abstract description 26
- 239000010802 sludge Substances 0.000 claims abstract description 25
- 244000005700 microbiome Species 0.000 claims abstract description 21
- 230000008569 process Effects 0.000 claims abstract description 21
- 239000003549 soybean oil Substances 0.000 claims abstract description 20
- 235000012424 soybean oil Nutrition 0.000 claims abstract description 20
- 235000011187 glycerol Nutrition 0.000 claims abstract description 19
- 235000019710 soybean protein Nutrition 0.000 claims abstract description 19
- 239000006041 probiotic Substances 0.000 claims abstract description 16
- 235000018291 probiotics Nutrition 0.000 claims abstract description 16
- 238000004945 emulsification Methods 0.000 claims abstract description 13
- 235000013305 food Nutrition 0.000 claims abstract description 12
- 230000000529 probiotic effect Effects 0.000 claims abstract description 12
- 230000036541 health Effects 0.000 claims abstract description 11
- 239000003814 drug Substances 0.000 claims abstract description 9
- 239000000126 substance Substances 0.000 claims description 29
- 238000002156 mixing Methods 0.000 claims description 21
- 239000003921 oil Substances 0.000 claims description 21
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- 238000004519 manufacturing process Methods 0.000 claims description 9
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- 239000007921 spray Substances 0.000 claims description 4
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- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 241000186000 Bifidobacterium Species 0.000 claims description 2
- 241000193171 Clostridium butyricum Species 0.000 claims description 2
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- 239000002245 particle Substances 0.000 description 19
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- 238000012360 testing method Methods 0.000 description 13
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 11
- 238000009826 distribution Methods 0.000 description 10
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- 108090000623 proteins and genes Proteins 0.000 description 8
- 230000008859 change Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 6
- 235000010469 Glycine max Nutrition 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000003674 animal food additive Substances 0.000 description 6
- 239000012153 distilled water Substances 0.000 description 6
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- 244000068988 Glycine max Species 0.000 description 4
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- 125000001165 hydrophobic group Chemical group 0.000 description 4
- 238000011056 performance test Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 102000057297 Pepsin A Human genes 0.000 description 3
- 108090000284 Pepsin A Proteins 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000000654 additive Substances 0.000 description 3
- 230000000996 additive effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 239000008187 granular material Substances 0.000 description 3
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- 229940111202 pepsin Drugs 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 102000004882 Lipase Human genes 0.000 description 2
- 108090001060 Lipase Proteins 0.000 description 2
- 239000004367 Lipase Substances 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 101710123874 Protein-glutamine gamma-glutamyltransferase Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 230000002776 aggregation Effects 0.000 description 2
- 238000004220 aggregation Methods 0.000 description 2
- 238000009395 breeding Methods 0.000 description 2
- 230000001488 breeding effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 102000038379 digestive enzymes Human genes 0.000 description 2
- 108091007734 digestive enzymes Proteins 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 235000019421 lipase Nutrition 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000002068 microbial inoculum Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
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- 238000005070 sampling Methods 0.000 description 2
- 230000003068 static effect Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 206010010774 Constipation Diseases 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 244000061121 Rauvolfia serpentina Species 0.000 description 1
- 108010046685 Rho Factor Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 240000001417 Vigna umbellata Species 0.000 description 1
- 235000011453 Vigna umbellata Nutrition 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000002457 bidirectional effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000019711 black bean protein Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000011162 core material Substances 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007071 enzymatic hydrolysis Effects 0.000 description 1
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- 239000000284 extract Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
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- 238000004108 freeze drying Methods 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
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- 238000011065 in-situ storage Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000019707 mung bean protein Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
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- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 229940071440 soy protein isolate Drugs 0.000 description 1
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- 239000004094 surface-active agent Substances 0.000 description 1
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Classifications
-
- 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
- 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
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- 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
- A23K20/158—Fatty acids; Fats; Products containing oils or fats
-
- 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
- A23K20/163—Sugars; Polysaccharides
-
- 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
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/035—Organic compounds containing oxygen as heteroatom
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/05—Organic compounds containing phosphorus as heteroatom
-
- 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
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
- A23L29/35—Degradation products of starch, e.g. hydrolysates, dextrins; Enzymatically modified starches
-
- 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
- A23L33/14—Yeasts or derivatives thereof
-
- 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
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Biotechnology (AREA)
- Physiology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- General Preparation And Processing Of Foods (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention provides a liquid microcapsule system, a preparation method and application thereof. The microcapsule system is mainly composed of stable emulsion formed by uniformly dispersing liquid microcapsule, and the liquid microcapsule is mainly composed of, by mass, 0.5-1.2 parts of microbial bacterial sludge, 1-3 parts of enzymolysis soybean protein, 2-8 parts of concentrated phospholipid, 8-10 parts of soybean oil, 2-10 parts of maltodextrin, 0.8-1.2 parts of glycerin and 6-25 parts of water. The invention also provides a preparation method of the microcapsule system, which comprises the step of carrying out vacuum emulsification treatment on the raw materials. The invention also provides an application of the liquid microcapsule in the processing process of products containing microorganisms, such as health care products, foods, probiotic medicaments or probiotic feeds. The liquid microcapsule system has the characteristics of high survival rate, good heat resistance, good slow release property, low cost and the like, and the preparation method is simple, and the survival quantity of thalli in processing and application is improved by spraying after coating, so that the application and processing cost is reduced.
Description
Technical Field
The invention belongs to the field of microbial additive application and processing, and particularly relates to a liquid microcapsule system, a preparation method and application thereof.
Background
The microbial feed additive is a feed additive approved by the agricultural rural area of the people's republic of China, and plays a beneficial role by improving the ecological balance of the intestinal flora of animals so as to improve the health level of the animals, disease resistance and digestion capacity. Is an effective means for effectively solving the problems of disease flooding, pathogen drug resistance, immunocompetence reduction, survival rate reduction and cultivation benefit reduction. The microbial preparation has the advantage that other medicines cannot be replaced, namely, the effects of disease treatment, disease prevention without disease and health care without disease. Even healthy animals can use the composition to improve the health level, and has bidirectional regulating effect on diarrhea and constipation.
The existing microbial feed additive is mainly powder or granule, and the production steps mainly comprise: extracting strains, culturing and breeding the strains in an enlarged manner, dehydrating finished products, mixing and packaging the products and the like (Sun Xiaoming. Production and application of microbial feed additives [ J ]. 04 th stage of culture and feed 2013: p 45-46); wherein, the microorganism cultured in the fermentation tank is processed by a finished product dehydration step to obtain the powder or granule of the microorganism. Therefore, in the microbial feed processing process, the microbial feed additive is mixed with other raw materials in the form of powder or granule to prepare feed, and thus, there are the following problems: 1. in the production process of the microbial additive, drying equipment is needed, and the problem of high production cost exists; 2. the problem of uneven distribution of microbial feed additives; 3. microorganisms have problems of poor acid resistance and heat resistance, low survival rate and poor colonisation ability in intestinal tracts.
For this reason, chinese patent application CN 113832047A discloses a TGase cross-linking modified lactobacillus reuteri microcapsule and a preparation method thereof, the lactobacillus reuteri microcapsule mainly uses concentrated lactobacillus reuteri bacterial liquid as a core material, uses fishskin gelatin and maltodextrin as wall materials, and the wall materials are prepared by TGase cross-linking modification treatment; wherein the Lactobacillus reuteri is Lactobacillus reuteri NCUF203.1. The preparation method disclosed by the patent application is simple and convenient to operate, low in energy consumption and high in viable bacteria survival rate, and provides a good method for preparing lactobacillus reuteri microcapsules by a spray drying method and reducing the production cost of lactobacillus reuteri preparations. The microcapsule prepared by the invention has the advantages that the number of the live bacteria of the embedded lactobacillus reuteri is greatly increased compared with that of the microcapsule prepared by uncrosslinked fishskin gelatin, and the microcapsule has good gastric acid resistance, enteric solubility and storage stability.
However, although the patent application discloses that the lactobacillus reuteri microcapsules still need drying treatment in the preparation process, the lactobacillus reuteri microcapsules are still unevenly distributed in the processing process of feeds, foods and the like, and the problems of high production cost and the like still exist; it is also difficult to solve the problems that microorganisms are sensitive to adverse factors such as high temperature and pressure, and are easily affected to inactivate bacteria during processing and storage.
Disclosure of Invention
In view of the above, the present invention aims to prepare a liquid microcapsule system, which uses microbial slime, enzymatic soy protein, concentrated phospholipid, soybean oil, maltodextrin, glycerol and the like as raw materials, and is prepared by an emulsification method under a vacuum state, so that the microorganisms in the liquid microcapsule system have high survival rate and embedding rate, and thus, the stability and colonization ability of the microorganisms in intestinal tracts are improved, and the subsequent combined post-spraying process is used for processing feeds, so that the problem of low survival rate caused by the influence of external factors such as pressure in the processing process of products containing the microorganisms is avoided; thereby reducing the production cost of processing the liquid microcapsule system and the microorganism-containing product.
The invention also aims to provide a preparation method of the liquid microcapsule system, which mainly comprises the steps of mixing raw materials in a vacuum state and preparing the liquid microcapsule system by combining an emulsifying method, and has the advantages of simple preparation method and low cost.
The invention also aims to provide an application of the liquid microcapsule system, wherein the liquid microcapsule system is added into microorganism-containing products such as health-care products, foods, probiotic medicaments, probiotic feeds and the like by adopting a post-spraying process, so that the liquid microcapsule system is distributed more uniformly in the microorganism-containing products, the survival rate of microorganisms in the microorganism-containing products is improved, the addition amount of the microcapsule system in the microorganism-containing products can be relatively reduced, and the cost is further reduced.
In order to achieve the above purpose, the present invention protects the following technical scheme:
the liquid microcapsule system mainly comprises, by mass, 0.5-1.2 parts of microbial bacterial sludge, 1-3 parts of enzymolysis soybean protein, 2-8 parts of concentrated phospholipid, 8-10 parts of soybean oil, 2-10 parts of maltodextrin, 0.8-1.2 parts of glycerin and 6-25 parts of water.
Preferably, the liquid microcapsules have a particle size distribution in the range of 1500-3000 nm, such as 1500 nm, 1600nm, 1700 nm, 1800 nm, 1900 nm, 2000nm, 2100 nm, 2200 nm, 2300 nm, 2400nm, 2500 nm, 2800 nm, 3000 nm, etc.; the particle size of the liquid microcapsules is more preferably distributed between 1800 and 2400mm. The mass percentage of the concentrated phospholipid in the liquid microcapsule system is preferably 5-20%. The mass percentage of the maltodextrin in the liquid microcapsule system is preferably 5-25%.
Preferably, the liquid microcapsule system comprises, by mass, 40 parts of microbial sludge 1 part, enzymatic hydrolysis of soybean protein 2 parts, concentration of phospholipid 3-7 parts, soybean oil 10 parts, maltodextrin 2-10 parts, glycerol 1 part and water in balance. The microbial bacterial sludge is preferably probiotic bacterial sludge, such as one or more of bacillus bacterial sludge, lactobacillus bacterial sludge, saccharomycete bacterial sludge, clostridium butyricum bacterial sludge, bifidobacterium bacterial sludge and the like; more preferably, the microbial sludge is lactobacillus reuteri sludge.
The concentrated phospholipid is preferably liquid plant concentrated phospholipid, and more preferably liquid soybean concentrated phospholipid.
The enzymolysis soybean protein is preferably enzymolysis soybean protein, enzymolysis black bean protein, enzymolysis red bean protein or enzymolysis mung bean protein, etc.
The preparation method of the liquid microcapsule system comprises the steps of mixing and emulsifying 0.5-1.2 parts of microbial sludge, 1-3 parts of enzymolysis soybean protein, 2-8 parts of concentrated phospholipid, 8-10 parts of soybean oil, 2-10 parts of maltodextrin, 0.8-1.2 parts of glycerol and 6-25 parts of water in a vacuum state to form a plurality of uniformly dispersed liquid microcapsules, and obtaining stable emulsion.
The preparation method comprises the following steps:
preparing an oil phase substance, uniformly mixing concentrated phospholipid and soybean oil at 40-60 ℃ to obtain the oil phase substance;
preparing a water phase substance, and uniformly mixing maltodextrin, glycerol and a part of water to obtain the water phase substance;
the preparation method comprises the steps of preparing a capsule system, uniformly mixing the microbial bacterial mud, the enzymolysis soy protein and the rest water, and then adding the oil phase substance and the water phase substance for emulsification treatment to form stable emulsion.
In accordance with the above, the step of preparing a capsule system comprises: firstly, mixing the microbial bacterial sludge, the enzymolysis soy protein and the rest water for 15-45 min in a vacuum state to form mixed bacterial liquid; and adding the oil phase substance and the water phase substance into the mixed bacterial liquid, and emulsifying for 60-120 min at room temperature under a vacuum condition to form a stable emulsion, thus obtaining the liquid microcapsule system.
Preferably, in the preparation process of the liquid microcapsule system, each step is stirred and mixed at a rotating speed of 500-1000 r/min.
The application of the liquid microcapsule system in processing products containing microorganisms such as health care products, foods, probiotics medicaments or probiotics feeds. Wherein the liquid microcapsule system is used as an additive in health products, foods and probiotic feeds.
Based on the application, the liquid microcapsule system is applied to the processing process of health products, foods, probiotics medicaments or probiotics feeds by adopting a post-spraying process.
Preferably, the temperature of the post-spraying process is room temperature, and the spraying pressure of the spraying process is less than or equal to 0.7 MPa (7 Bar) of the surface pressure of the spraying machine. The gauge pressure of the spraying machine of the post-spraying process is preferably 0.2-0.6 MPa.
Therefore, the liquid microcapsule system provided by the invention is a stable milky liquid formed by taking microbial sludge, soybean protein, concentrated phospholipid, soybean oil, maltodextrin, glycerol and water as raw materials and performing emulsification treatment in a vacuum state; in the microcapsule system, soy protein is mainly used as a protective agent for microorganisms; maltodextrin is mainly used as a protective agent and an emulsifying agent of microorganisms; the concentrated phospholipid material is mainly used as an emulsion stabilizer of the liquid microcapsule system, hydrophobic groups and protein hydrophobic groups of the concentrated phospholipid material can better wrap soybean oil drops, and hydrophilic groups and protein hydrophilic groups of the concentrated phospholipid material can better extend into an aqueous phase mainly formed by maltodextrin, glycerol and water, so that a stable structure of emulsion is formed; thus, the liquid microcapsule has better embedding effect, and bacteria in the liquid microcapsule are detectedThe activity can reach 10 9 CFU/mL, the embedding rate can reach 99.11%. The liquid microcapsule has good heat resistance and slow release property, and improves the tolerance of lactobacillus reuteri to adverse factors such as digestive enzymes and the like. Therefore, the liquid microcapsule system is easy to digest and obtain, and can be widely applied to the fields of probiotic health products, foods, medicaments, feeds and the like.
The preparation method of the liquid microcapsule system provided by the invention comprises the steps of firstly compounding bacterial mud with enzymolysis soy protein, and then carrying out emulsification reaction with the prepared oil phase and water phase under vacuum; compared with the existing preparation method of the probiotics coating material, the preparation method is simpler, the cost is lower, and the effect is better. In addition, the raw materials used for the liquid microcapsule prepared by the method provided by the invention are easy to obtain and are common food materials, and the prepared liquid microcapsule is stable and easy to store.
Therefore, the liquid microcapsule thalli provided by the invention has the characteristics of high survival rate, good heat resistance, good slow release property, low cost and the like, and the preparation method is simple, and the survival number of thalli in processing and application is improved by spraying after coating, so that the application and processing cost is reduced; the problems that the existing microorganism is sensitive to adverse factors such as high temperature, pressurization and the like and is easy to be influenced in the processing and storage processes to inactivate bacteria are also solved.
Drawings
FIG. 1 is a photograph of a phospholipid, wherein the left photograph is a photograph of a modified phospholipid, and the right photograph is a photograph of a concentrated phospholipid.
Fig. 2 is a graph showing the effect of phospholipids on lactobacillus reuteri liquid microcapsules, wherein graph a: ESI, EAI embedding rate, graph B: potential, graph C: average particle size, panel D: particle size distribution and graph E 1 -E 4 : microscopic observation of the figures, and significant differences between the data containing different lowercase letters in the subscripts in figures A-CP＜ 0.05）。
Fig. 3 is an external view of lactobacillus reuteri liquid microcapsule, wherein, fig. a: photo of directly prepared original liquid microcapsule, panel B: photograph of 10-fold diluted liquid microcapsule, panel C: photographs of liquid microcapsules diluted 100-fold, panels D and E: and (5) microscopic observation.
Fig. 4 is a graph showing the effect of maltodextrin on lactobacillus reuteri liquid microcapsules, wherein graph a: ESI, EAI embedding rate, graph B: potential, graph C: average particle size, panel D: particle size distribution and graph E 1 -E 4 : microscopic observation of the figures, and significant differences between the data containing different lowercase letters in the subscripts in figures A-CP＜ 0.05）。
Fig. 5 is a graph showing the effect of emulsification time on lactobacillus reuteri liquid microcapsules, wherein graph a: ESI, EAI embedding rate, graph B: potential, graph C: average particle size, panel D: particle size distribution and graph E 1 -E 4 : microscopic observation of the figures, and significant differences between the data containing different lowercase letters in the subscripts in figures A-CP＜ 0.05）。
Fig. 6 is a graph of the effect of temperature on lactobacillus reuteri microcapsules, wherein graph A, B, C: graph E of the Effect of bacterial liquid colony count 1 -E 4 : microscopic observation of the figures, and significant differences between the data containing different lowercase letters in the subscripts in figures A-CP＜ 0.05）。
Fig. 7 is a micrograph of lactobacillus reuteri microcapsules in gastrointestinal fluids at different digestion times: microscopic pictures under 40 x mirror in gastric juice digestion (A 1 -A 5 ) And a picture under a 100-fold mirror (C) 1 -C 5 ) The method comprises the steps of carrying out a first treatment on the surface of the Microscopic pictures under 40-fold mirror in intestinal juice digestion (B 1 -B 5 ) And a picture under a 100-fold mirror (D 1 -D 5 ）。
Fig. 8 is a photograph and a microscopic image of the lactobacillus reuteri liquid microcapsule system provided in the embodiment of the present invention before and after spraying.
Fig. 9 is a bar chart of total bacterial colony count before and after spraying of lactobacillus reuteri liquid microcapsule system provided by the embodiment of the invention.
FIG. 10 is a graph showing the effect of different spray pressures on the activity of the liquid microcapsule system of Lactobacillus reuteri provided in the examples of the present invention.
Detailed Description
The technical scheme of the invention is further described in detail through the following specific embodiments.
The liquid microcapsule system provided by the invention mainly takes bacterial mud, enzymolysis soybean protein, concentrated phospholipid, soybean oil, maltodextrin, glycerol and water as raw materials, and is subjected to vacuum emulsification to form uniform and stable emulsion.
The invention also provides a preparation method of the liquid microcapsule system, which comprises the following steps:
preparing an oil phase substance, slightly heating and uniformly mixing concentrated phospholipid and soybean oil to obtain the oil phase substance;
preparing a water phase substance, and uniformly mixing maltodextrin and glycerol to obtain the water phase substance;
preparing a capsule system, firstly mixing microbial bacterial mud and enzymolysis soy protein for 15-45 min in a vacuum state to form mixed bacterial liquid; and adding the oil phase substance and the water phase substance into the mixed bacterial liquid, and emulsifying for 60-120 min at room temperature under a vacuum condition to form a stable emulsion, thus obtaining the liquid microcapsule system.
The main reagents and materials used in the following examples of the present invention are shown in Table 1.
TABLE 1 Main reagents and materials
| Principal materials and reagents | Manufacturing factories |
| MT476913 Lactobacillus reuteri | Industrial microorganism strain preservation and breeding engineering laboratory (Henan Zhengzhou) |
| Soybean | Zhengzhou Yonghui park metallocene supermarket purchase |
| Phospholipid | Zhengzhou four-dimensional science and technology Co Ltd |
| Goldlong fish soybean oil | Sea-benefiting grain and oil industry Co.Ltd |
| Maltodextrin | Soy Biotechnology Co.Ltd |
| Pepsin (10000 NFU/mg) | Bioengineering (Shanghai) Co., Ltd. |
| Lipase (Activity 20000U/g) | Soy Biotechnology Co.Ltd |
| Trypsin (10000 NFU/mg) | Bioengineering (Shanghai) Co., Ltd. |
The principal configuration solutions and materials employed in the following examples of the present invention are as follows:
1) MRS liquid medium: peptone 10g, beef extract 10g, yeast extract 5 g, glucose 20 g, sodium acetate 5 g, tween-80 ml, magnesium sulfate 0.2 g, manganese sulfate 0.05 g, dipotassium hydrogen phosphate 2 g, C 6 H 14 N 2 O 7 2 g. Deionized water 1000 and ml, regulating pH to 6.2-6.4 with hydrochloric acid, and sterilizing at 121deg.C for 20 min.
2) MRS solid medium: the MRS liquid culture medium obtained in the above 1) is added with 18 g agar, and sterilized at 121 ℃ for 20 min.
3) The preparation method of the bacterial mud comprises the following steps: activating strain, adding 1 mL Lactobacillus reuteri strain into 100 mL MRS liquid culture medium, standing at 37.0deg.C for culturing 14: 14 h for first activationThe method comprises the steps of carrying out a first treatment on the surface of the Adding 3 mL of first activated lactobacillus reuteri bacteria liquid into 100 mL of MRS liquid culture medium, and performing stationary culture at 37.0 ℃ for 14 h for second activation; inoculating 4 mL second activated lactobacillus reuteri bacteria liquid into 100 mL MRS liquid culture medium, standing at 37.0deg.C for 14 hr, centrifuging at 5000r/min for 10 min, collecting precipitate to obtain lactobacillus reuteri bacteria mud with viable count of 10 10 CFU/mL。
4) The preparation method of the enzymolysis soybean protein comprises the following steps:
preparation of soy protein isolate (soybean protein isolate, SPI): cleaning soybean, removing impurities such as stones, crushing, sieving with a 80-mesh sieve, removing fat by petroleum ether, mixing defatted soybean powder with distilled water at a ratio of 1:10 (w/w), stirring 1 h, adjusting pH with 2 mol/L NaOH to 8.0, centrifuging at 50deg.C in water bath for 1 h,5000r/min, centrifuging at 4.8,4 ℃ with 2 mol/L HCL for the supernatant, standing overnight, centrifuging at 5000r/min for 50 min the next day, taking out precipitate, mixing with distilled water, adjusting pH to 7.0, and freeze-drying to obtain SPI;
preparation of enzymatically hydrolyzed soy protein (enzymatic hydrolysate of soybean protein isolate, ESP): preparing 3% SPI solution, regulating pH to 2.0, stirring at 37deg.C for 10 min, adding pepsin (enzyme-substrate ratio 1:100), performing enzymolysis, keeping pH constant, stopping reaction after 2. 2 h, regulating pH to 7.0, inactivating enzyme, and lyophilizing to obtain enzymolysis soybean protein.
5) Preparation of simulated gastric fluid and intestinal fluid
Simulating gastric juice, weighing 10g sodium chloride, adding distilled water for dissolution, adjusting the pH to 1.5 by using 1.5 mol/L HCl, transferring into a 500 mL volumetric flask, adding 0.2 g pepsin to a constant volume, and preparing in situ.
Simulating intestinal juice, weighing 6.8 of g of dipotassium hydrogen phosphate, adding distilled water for dissolution, adjusting the pH to 7.4 by using 1 mol/L NaOH, transferring into a 1000 mL volumetric flask, adding 2 g trypsin and 2 g lipase to a constant volume, and preparing the intestinal juice on site.
The proportion of raw materials, preparation process parameters and using method in the liquid microcapsule system provided by the invention have important influence on the processing performance, and lactobacillus reuteri is taken as an example to further explain the invention.
The liquid microcapsule system provided by the following examples of the invention is 40 g, and is mainly prepared by the following steps:
preparing an oil phase substance, uniformly mixing 3-7 g concentrated phospholipid and 10g soybean oil at 40-60 ℃ at a rotating speed of 1000 r/min to obtain the oil phase substance;
preparing a water phase substance, dissolving 2-10 g maltodextrin in a part of water to form a maltodextrin solution, and uniformly mixing with 1g glycerin to obtain the water phase substance;
preparing a capsule system, firstly mixing 2 g soy protein and the rest water in a flat-bottom flask to form a soy protein solution, then adding 1g lactobacillus reuteri bacteria mud, and stirring for 20min in vacuum at 1000 r/min; then adding the oil phase substance and the water phase substance, and stirring and emulsifying at room temperature under vacuum (-0.1 MPa) to form lactobacillus reuteri liquid microcapsule to obtain 40 g stable emulsion: a liquid microcapsule system.
The performance test method adopted by each embodiment of the invention is as follows:
(1) Determination of Emulsion Activity Index (EAI) and Emulsion Stability Index (ESI)
The lactobacillus reuteri liquid microcapsule is diluted by 1000 times with 0.10 percent sodium dodecyl sulfate, the blank group is zeroed with 0.10 percent sodium dodecyl sulfate, the measurement is carried out at the wavelength of 500 nm, the light absorption values at 0min and 30 min are recorded, and the light absorption values are respectively recorded as A 0 With A3 0 . The absorbance is between 0.2 and 0.8. Emulsion Stability Index (ESI), emulsion Activity Index (EAI) were calculated:
wherein: t is a constant of 2.303, N is a dilution factor, ρ is the emulsion protein concentration, and φ is the oil quality fraction.
(2) Particle size and Zeta potential measurement
Diluting lactobacillus reuteri liquid microcapsule sample with distilled water 1000 times, taking out 1 mL, and measuring the particle size and Zeta potential of the microcapsule by using a Markov particle size meter and a Zeta potential meter at 25 ℃.
(3) Determination of the embedding Rate
Diluting lactobacillus reuteri microcapsule system sample 100 times, and performing colony count to N 0 The samples were then diluted appropriately and centrifuged to remove the supernatant, and colony counts were counted as N by plate counting.
Embedding ratio (ME):。
(4) Test of Heat resistance
Liquid microcapsules of 1g lactobacillus reuteri were diluted 10-fold with distilled water and treated at 70 ℃, 85 ℃ and 100 ℃ with 30 s, 45 s and 60 s, respectively. Viable counts were performed by the gradient dilution plate method, 3 replicates per treatment, with lactobacillus reuteri solution as a control.
(5) In vitro simulated digestion test
Static digestion, simulated gastric fluid and lactobacillus reuteri liquid microcapsules were added to a triangular flask, shaken for 2 h in a 37 ℃ water bath of 110 r/min, sampled every 30 min and immediately assayed for relevant indicators. And (3) immediately after the gastric juice sampling is finished, adding simulated intestinal juice, wherein the time and the method for sampling the intestinal juice are the same as those of the simulated gastric juice. And simultaneously performing a comparison experiment.
(6) Microscopic observation
The lactobacillus reuteri liquid microcapsule was diluted 10-fold, observed with an optical microscope, and then the sample was coated and fixed, dyed by a gram dyeing method, crystal violet dyed for 1 min, iodized liquid mordant dyed for 1 min,95% ethanol decolorized for 30 s, and tsar counterstained for 1 min, and photographed under 16×40 and 16×100 (oil glasses).
1. Effect of concentrated phospholipids on liquid microcapsule systems
Phospholipids are natural surfactants having hydrophilic and lipophilic groups, can be used as emulsifier stabilizers, binders and lubricants, and are safe, inexpensive and readily available. However, as shown in FIG. 1 (left panel), the modified phospholipids are easily layered and yellow in color at normal temperature; the concentrated phospholipid is a uniform solution without layering at normal temperature as shown in figure 1 (right graph); therefore, the present invention selects the concentrated phospholipid as an emulsion stabilizer to ensure that the liquid microcapsule system provided by the present invention is a stable emulsion in which the liquid microcapsules are uniformly dispersed. Under otherwise identical conditions, the amount of concentrated phospholipid added has a significant impact on the performance of the liquid microcapsule system.
Test conditions: maltodextrin 4 g, concentrated phospholipid, 3 g, 4 g, 5 g, 6 g, 7 g, lactobacillus reuteri 1g, enzymolysis soybean protein 2 g, soybean oil 10g, glycerin 1g and water in sequence; according to the preparation method, the corresponding liquid microcapsule systems are prepared according to the formula, and the stirring and emulsifying time of each liquid microcapsule system in the preparation process is 90 minutes. The results of the performance test of each lactobacillus reuteri liquid microcapsule are shown in fig. 2.
As can be seen from fig. 2: the emulsion stability, the emulsion activity and the embedding rate are obviously changed along with the increase of the concentration of the concentrated phospholipid, and the emulsion is increased and then reducedP<0.05). At a concentrated phospholipid content of 6 g, the emulsion stability value, the emulsion activity value and the entrapment rate were all maximized. Along with the increase of the content of the concentrated phospholipid, the emulsion stability embedding rate of the system is continuously improvedP<0.05），E 1 Ratio E 2 Dense and many spheres E 3 Ratio E 4 The bacterial mass is large. The concentration of the phospholipid is obviously reduced when 7 g is addedP<0.05). Because the concentrated phospholipid is an emulsifier, the protein and the concentrated phospholipid in the liquid microcapsule system are both amphiphilic, the oil drops can be better coated by the hydrophobic groups of the phospholipid and the hydrophobic groups of the protein, and the hydrophilic groups of the phospholipid and the hydrophilic groups of the protein can be better extended into the water phase, so that a stable structure of emulsion is formed, the interfacial tension of water and oil is reduced, but when the concentrated phospholipid reaches a certain concentration, the combination of the protein and the concentrated phospholipid reaches saturation, the emulsifying property of the emulsion tends to be mild, and even the stability is reduced; the excessive addition of the concentrated phospholipid leads to the formation of emulsion drops with different sizes in a compound system, and the emulsion drops are unevenly distributed and are easy to migrate, flow and the like. The particle size decreased and increased with increasing content of concentrated phospholipid, with two major peaks at 3 g, 5 g, 7 g, relatively short and broad at 4 g and relatively high and narrow at 6 g, indicating relatively uniform emulsion droplet distribution.
Wherein the lactobacillus reuteri liquid microcapsule formed when the added amount of the concentrated phospholipid was 6 g is shown in fig. 3. The lactobacillus reuteri liquid microcapsule is formed by embedding lactobacillus reuteri by wall material phospholipid, maltodextrin and the like, and is milky white liquid as shown in fig. 3 (A); the microcapsule shown in the figure (A) is diluted 10 times to be white liquid shown in the figure (B), the microcapsule is spherical (D) with uniform size under a 40 times microscope, and the microcapsule is observed to be a thallus aggregation (E) under a 100 times oil microscope, so that the liquid microcapsule wraps the thallus, a barrier is formed by a wall material outside the thallus, the thallus is isolated from the external environment, and the stability and activity of the thallus are enhanced. And (C) is a photo drawing of 100 times of microcapsule dilution shown in the drawing (A), and is mainly used for measuring various performance indexes of the lactobacillus reuteri liquid microcapsule.
2. Influence of maltodextrin on liquid microcapsule systems
Under otherwise identical conditions, the amount of maltodextrin added has a significant effect on the performance of the liquid microcapsule system.
Test conditions: maltodextrin is 2 g, 4 g, 6 g, 8 g, 10g, concentrated phospholipid 6 g, lactobacillus reuteri 1g, enzymolysis soybean protein 2 g, soybean oil 10g, glycerin 1g and the balance of water in sequence, and the concentration of maltodextrin in the whole liquid microcapsule system is 5%, 10%, 15%, 20% and 25% in sequence through calculation; according to the preparation method, the corresponding liquid microcapsule systems are prepared according to the formula, and the stirring and emulsifying time of each liquid microcapsule system in the preparation process is 90 minutes. The results of the performance test of each lactobacillus reuteri liquid microcapsule are shown in fig. 4.
As can be seen from fig. 4: as the concentration of maltodextrin increases, the EAI and ESI are both increased as the concentration of maltodextrin increasesP<0.05 When the concentration of maltodextrin increases to 15%, both EAI and ESI decrease with increasing concentration of maltodextrinP<0.05). The embedding rate is increased firstly and then decreased as the concentration of maltodextrin is increasedP<0.05 The embedding rate was highest when the maltodextrin concentration reached 15%; e (E) 1 Ratio E 2 Dense and many spheres E 3 Ratio E 4 The bacterial cells are agglomerated. When the maltodextrin concentration reached 20% and 25%It is possible that maltodextrin has a strong interaction with the emulsifier phospholipid, which affects the stability of the microcapsules and thus the entrapment effect. The absolute value of the potential is firstly reduced and then increased along with the increase of the concentration of maltodextrinP<0.05 The absolute value is at a minimum at a maltodextrin concentration of 15%. The particle size is not significantly different between the maltodextrin concentration of 5% and 10%P<0.05 Particle size is reduced when maltodextrin concentration reaches 15%P<0.05 Particle size gradually increases with increasing concentrationP<0.05 A) is provided; the average particle size distribution at 5% and 10% concentration was narrow and low, the peaks at 20% and 25% concentration were both low and broad, the peak at 15% concentration was one peak at maximum and narrow, indicating that the maltodextrin concentration was 15%, i.e., the maltodextrin addition was 6 g, and the liquid droplet distribution of the lactobacillus reuteri liquid microcapsule emulsion was relatively uniform.
3. Influence of the emulsion time on the liquid microcapsule system
The emulsifying time of the liquid microcapsule system has an important influence on the performance of the liquid microcapsule system under the same other conditions.
Test conditions: maltodextrin 6 g, concentrated phospholipid 6 g, lactobacillus reuteri 1g, enzymolysis soybean protein 2 g, soybean oil 10g, glycerin 1g and the balance of water; the raw materials are respectively stirred and emulsified for 30 min, 60 min, 90min, 120min and 150 min at room temperature under vacuum state, so as to obtain different liquid microcapsule systems. The results of the performance test of the respective lactobacillus reuteri liquid microcapsule systems are shown in fig. 5.
As can be seen from fig. 5: the EAI and the ESI gradually rise with the increase of the emulsification timeP<0.05 Maximum when the emulsification time reaches 90min, and no obvious change of EAI and ESI when the emulsification time is increasedP<0.05). The embedding rate is increased and then decreased along with the increase of the emulsification timeP<0.05 The embedding rate is maximum when the emulsifying time reaches gradually increasing 90min from 30 to 90min, and the embedding rate is reduced when the emulsifying time exceeds 90minP<0.05 Fig. E 1 Comparison E 2 Dense and many spheres, fig. E 3 Comparison E 4 The bacterial cells have large aggregation and canThe magnetic stirring can destroy the embedding system, break the interaction between maltodextrin and emulsifier phospholipid, reduce emulsion breaking caused by emulsion stability, increase free thallus in emulsion and reduce embedding rate. The average particle size is narrow and short in peak of the graph of 30 min, the peak ratio of the graph of 60 min is 90min and the peak height of 150 min is wide, two peaks of 120min indicate that the particle size distribution of the emulsion is uneven, and the peak of 90min is sharp and narrow, which indicates that the emulsion distribution is even.
Examples
The embodiment provides a liquid microcapsule system, namely maltodextrin 6 g, concentrated phospholipid 6 g, lactobacillus reuteri 1g, enzymolysis soybean protein 2 g, soybean oil 10g, glycerin 1g and the balance of water; the raw materials are stirred and emulsified for 90min at room temperature under vacuum state. Through detection, the bacterial activity of the liquid microcapsule system reaches 10 9 CFU/mL, the embedding rate reaches 99.11%. The heat resistance test results, in vitro digestion test results and processing application test results of the liquid microcapsule system are shown in fig. 6-10.
(1) Results of the Heat resistance test
As can be seen from fig. 6: the total number of the bacterial colonies of the control group is obviously reduced along with the lengthening of the treatment time at 70 DEG CP<0.05 The total number of colonies of the microcapsule group is also obviously reduced along with the lengthening of the treatment timeP<0.05 And the total number of colonies of the control group and the microcapsule group is not very large, the control group is bacterial liquid, and the bacterial liquid contains MRS culture medium and has a certain protection effect on bacterial cells. At 85 ℃, along with the lengthening of the treatment time, the control group and the microcapsule group have no obvious change, but the total number of colony of the microcapsule group is obviously more than that of the control group, which indicates that the coating wall material has certain heat resistance. The change of the control group and the microcapsule group along with the lengthening of the treatment time at 100 ℃ is not obvious, but the survival number of the bacterial cells of the bacterial liquid control group is obviously more than that of the control group according to the total number of bacterial colonies of the microcapsule group in each treatment period, which proves that the coating wall material maltodextrin has good heat resistance.
(2) Results of in vitro digestion test
From FIG. 7（A 1 -A 5 ) It can be seen that: the microcapsule has no obvious change in the process of simulating gastric juice digestionP>0.05 No significant change after increasing in intestinal juiceP>0.05 A) is provided; the control bacterial liquid group is obviously reduced just after gastric juice is addedP<0.05 No significant changes in gastric juice and intestinal juiceP<0.05 A) is provided; the embedded wall material has a certain protection effect on lactobacillus reuteri, so that the lactobacillus reuteri is prevented from being damaged by a strong acid environment; when the microcapsule reaches intestinal juice, the intestinal juice is alkaline and the gastric juice is acidic to neutralize, so that the pH of the solution is increased to be suitable for the growth of the Rauwolfia, and the total bacterial colony number in the intestinal juice is more than that of the gastric juice; the bacterial liquid in the control group contains a liquid culture medium, the culture medium is a nutrient substance, meanwhile, the culture medium also provides nutrition for lactobacillus reuteri, and the pH in the intestinal liquid is increased to promote the growth and propagation of bacterial colonies so that the total number of bacterial colonies in the intestinal liquid is more than that in gastric juice.
From FIG. 7 (B) 1 -B 5 ) It can be seen that: with the increase of digestion time, the EAI and ESI are gradually reduced to intestinal juice and then the change is not obviousP>0.05 The pH rapidly decreases after gastric juice addition and the microcapsules are in a strong acid environment, decreasing stability and thus leading to a gradual decrease in both EAI and ESI.
From FIG. 7 (C 1 -C 5 ) Sum (D) 1 -D 5 ) It can be derived that: particle size is reduced in gastric juice along with the increase of digestion timeP<0.05 The microcapsule is cracked in the small intestine without obvious change in intestinal juice, and then secondary emulsification is carried out, so that the size is uniform; the absolute value of the potential value in the whole digestion gradually decreases with the increase of the digestion timeP<0.05）。
Static gastric juice digestion 2 h microcapsule group viable count is 1.26×10 7 CFU/mL, intestinal juice digestion 2 h microcapsule viable count is 2.16X10 8 CFU/mL, after 4 h is digested by gastrointestinal fluid, the viable count of the microcapsule is 1.85 multiplied by 10 8 CFU/mL; thus, it is demonstrated that the liquid microcapsules have a slow release effect in the intestinal tract.
(3) Influence of the pressure parameters of the processing procedure on the liquid microcapsule system
The existing microbial inoculum is interfered by factors such as pressure and the like in the processing and application processes, and has great influence on the activity of the microbial inoculum. In the processing process of the liquid microcapsule system provided by the embodiment of the invention by adopting a post-spraying process, the spraying pressure has an important influence on the embedding performance of the liquid microcapsule, and the specific influence is further explained through the following test.
1) Test conditions: photo graphs and colony counts of lactobacillus reuteri liquid microcapsule systems before and after spraying by using a gauge pressure of 3 bar of a spraying machine after diluting the liquid microcapsule by 100 times are shown in fig. 8 and 9.
As can be seen from fig. 8: after spraying the lactobacillus reuteri liquid microcapsule, the liquid drops of the capsule are smaller, and the thallus is still agglomerated; the following is explained: the pressure makes the dispersion of the liquid microcapsule system more uniform, and the coating is not destroyed; meanwhile, the sample of the liquid microcapsule system of the lactobacillus reuteri subjected to spray coating is whiter in color and more uniform in mixing.
As can be seen in fig. 9: the total colony count of the lactobacillus reuteri liquid microcapsule before and after spraying treatment is 10 9 The difference is not obvious, which indicates that the spraying has less influence on the survival of the thalli.
2) Test conditions: after diluting the lactobacillus reuteri liquid microcapsule provided in the embodiment by 100 times, spraying by using a spraying machine respectively through different gauge pressures. And then measuring and calculating the total number of bacterial colonies and the embedding rate of the diluted liquid microcapsule system after spraying treatment. The gauge pressure of the spraying machine is 2, 3, 4, 5 and 6 Bar (1 bar=100 kpa=0.1 MPa). The results are shown in FIG. 10.
As can be seen from fig. 10: as the pressure increases, the total number of colonies increases and decreases, the total number of colonies is the largest under 3 bar, but the total number of colonies is 10 under 2-6 bar 9 The lactobacillus reuteri liquid microcapsule has better pressure-resistant effect; the embedding rate is increased and then reduced along with the increase of the pressure, the pressure is 3 bar, and the embedding rate is the best and can reach 98 percent; embedding rate is more than 90 percent and even more than 96 percent under the pressure of 2-6 bar, which indicates embeddingThe effect is better. Therefore, the liquid microcapsule can protect microbial cells in the liquid microcapsule from being damaged by factors such as pressure and the like in the post-spraying processing process, and the survival number of the microbial cells in the processing process of health care products, foods, medicaments or feeds and the like is improved.
Therefore, the lactobacillus reuteri liquid microcapsule provided by the embodiment of the invention is milky white liquid, has good dispersibility, has uniform and dense spherical size under a microscope observation 40 times, and gathers thalli under a microscope 100 times; has good heat resistance and slow release property, improves the tolerance of lactobacillus reuteri to adverse factors such as digestive enzymes, and has the characteristics of high survival rate and embedding rate. Meanwhile, after embedding of the lactobacillus reuteri, post-spraying avoids the influence of external factors such as pressure and the like on the lactobacillus reuteri in the processing process, so that enough living bacteria enter the intestinal tract, the probiotic effect is further exerted, the application range of the lactobacillus reuteri is expanded, and a theoretical foundation is laid for the lactobacillus reuteri in food processing.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting the same; while the invention has been described in detail with reference to the preferred embodiments, those skilled in the art will appreciate that: modifications may be made to the specific embodiments of the present invention or equivalents may be substituted for part of the technical features thereof; without departing from the spirit of the invention, it is intended to cover the scope of the invention as claimed.
Claims (10)
1. The liquid microcapsule system is characterized by being a stable emulsion formed by uniformly dispersing liquid microcapsules, wherein the liquid microcapsule system mainly comprises, by mass, 0.5-1.2 parts of microbial bacterial sludge, 1-3 parts of enzymolysis soybean protein, 2-8 parts of concentrated phospholipid, 8-10 parts of soybean oil, 2-10 parts of maltodextrin, 0.8-1.2 parts of glycerol and 6-25 parts of water.
2. The liquid microcapsule system according to claim 1, wherein the mass percentage of the concentrated phospholipid in the liquid microcapsule system is 5-20%.
3. Liquid microcapsule system according to claim 1 or 2, characterized in that the maltodextrin is present in the liquid microcapsule system in a mass percentage of 5% -25%.
4. The liquid microcapsule system according to claim 3, wherein the liquid microcapsule system comprises, by mass, 40 parts of microbial sludge 1 part, enzymatic soybean protein 2 parts, concentrated phospholipid 3-7 parts, soybean oil 10 parts, maltodextrin 2-10 parts, glycerin 1 part, and the balance water.
5. The liquid microcapsule system according to claim 1, 2 or 4, wherein the microbial bacterial sludge is one or more of bacillus sludge, lactobacillus sludge, saccharomycete sludge, clostridium butyricum sludge, bifidobacterium sludge and the like.
6. The preparation method of the liquid microcapsule system comprises the steps of mixing and emulsifying 0.5-1.2 parts of microbial bacterial sludge, 1-3 parts of enzymolysis soybean protein, 2-8 parts of concentrated phospholipid, 8-10 parts of soybean oil, 2-10 parts of maltodextrin, 0.8-1.2 parts of glycerol and 6-25 parts of water in a vacuum state to form a plurality of uniformly dispersed liquid microcapsules, and obtaining stable emulsion.
7. The method of manufacturing according to claim 6, comprising the steps of:
preparing an oil phase substance, uniformly mixing concentrated phospholipid and soybean oil at 40-60 ℃ to obtain the oil phase substance;
preparing a water phase substance, and uniformly mixing maltodextrin, glycerol and a part of water to obtain the water phase substance;
the preparation method comprises the steps of preparing a capsule system, uniformly mixing the microbial bacterial mud, the enzymolysis soy protein and the rest water, and then adding the oil phase substance and the water phase substance for emulsification treatment to form stable emulsion.
8. The liquid microcapsule system according to claim 7, wherein the step of preparing a capsule system comprises: firstly, mixing the microbial bacterial sludge, the enzymolysis soy protein and the rest water for 15-45 min in a vacuum state to form mixed bacterial liquid; and adding the oil phase substance and the water phase substance into the mixed bacterial liquid, and emulsifying for 60-120 min at room temperature under a vacuum condition to form a stable emulsion, thus obtaining the liquid microcapsule system.
9. The application of a liquid microcapsule system in processing a microorganism-containing product, wherein the microorganism-containing product is a health product, a food, a probiotic medicament or a probiotic feed.
10. The use according to claim 9, wherein the liquid microcapsule system is applied in the processing of health products, food, probiotic medicaments or probiotic feeds by a post-spray process, and the spray pressure of the post-spray process is less than or equal to 0.7 MPa of the sprayer gauge pressure.
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