CN115399481B - Functional edible film loaded with probiotics and preparation method thereof - Google Patents
Functional edible film loaded with probiotics and preparation method thereof Download PDFInfo
- Publication number
- CN115399481B CN115399481B CN202210859329.8A CN202210859329A CN115399481B CN 115399481 B CN115399481 B CN 115399481B CN 202210859329 A CN202210859329 A CN 202210859329A CN 115399481 B CN115399481 B CN 115399481B
- Authority
- CN
- China
- Prior art keywords
- liposome
- protamine
- probiotics
- probiotic
- edible film
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 239000006041 probiotic Substances 0.000 title claims abstract description 90
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 90
- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- 239000002502 liposome Substances 0.000 claims abstract description 94
- 102000007327 Protamines Human genes 0.000 claims abstract description 60
- 108010007568 Protamines Proteins 0.000 claims abstract description 60
- 229940048914 protamine Drugs 0.000 claims abstract description 60
- 230000000529 probiotic effect Effects 0.000 claims abstract description 37
- 150000002632 lipids Chemical class 0.000 claims abstract description 32
- 239000000243 solution Substances 0.000 claims description 43
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 30
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 238000003756 stirring Methods 0.000 claims description 18
- 239000000725 suspension Substances 0.000 claims description 17
- 239000002904 solvent Substances 0.000 claims description 14
- 229920001661 Chitosan Polymers 0.000 claims description 13
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- 229920002494 Zein Polymers 0.000 claims description 11
- 238000010438 heat treatment Methods 0.000 claims description 11
- 239000005019 zein Substances 0.000 claims description 11
- 229940093612 zein Drugs 0.000 claims description 11
- 229960000583 acetic acid Drugs 0.000 claims description 10
- 239000003963 antioxidant agent Substances 0.000 claims description 10
- 230000003078 antioxidant effect Effects 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 8
- 239000012362 glacial acetic acid Substances 0.000 claims description 8
- 238000004132 cross linking Methods 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 230000000887 hydrating effect Effects 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- ZTHYODDOHIVTJV-UHFFFAOYSA-N Propyl gallate Chemical compound CCCOC(=O)C1=CC(O)=C(O)C(O)=C1 ZTHYODDOHIVTJV-UHFFFAOYSA-N 0.000 claims description 6
- 238000001035 drying Methods 0.000 claims description 6
- 238000001704 evaporation Methods 0.000 claims description 6
- 108060006613 prolamin Proteins 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 239000004014 plasticizer Substances 0.000 claims description 5
- -1 polytetrafluoroethylene Polymers 0.000 claims description 5
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 5
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 5
- 239000000843 powder Substances 0.000 claims description 5
- 239000002994 raw material Substances 0.000 claims description 5
- 238000009849 vacuum degassing Methods 0.000 claims description 5
- 230000001580 bacterial effect Effects 0.000 claims description 4
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- 239000000473 propyl gallate Substances 0.000 claims description 3
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- 235000010388 propyl gallate Nutrition 0.000 claims description 3
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- 239000003960 organic solvent Substances 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000000265 homogenisation Methods 0.000 claims 1
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- 238000000034 method Methods 0.000 abstract description 12
- 239000001257 hydrogen Substances 0.000 abstract description 3
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 3
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- 230000007547 defect Effects 0.000 abstract 1
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- 241000186000 Bifidobacterium Species 0.000 description 35
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 15
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 12
- 230000000052 comparative effect Effects 0.000 description 11
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- CITHEXJVPOWHKC-UUWRZZSWSA-N 1,2-di-O-myristoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCCCCCCC CITHEXJVPOWHKC-UUWRZZSWSA-N 0.000 description 8
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- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 230000002496 gastric effect Effects 0.000 description 6
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- 238000004220 aggregation Methods 0.000 description 5
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- 235000011187 glycerol Nutrition 0.000 description 5
- WTJKGGKOPKCXLL-RRHRGVEJSA-N phosphatidylcholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCCC=CCCCCCCCC WTJKGGKOPKCXLL-RRHRGVEJSA-N 0.000 description 5
- 238000001308 synthesis method Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 230000008020 evaporation Effects 0.000 description 4
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- 229920001282 polysaccharide Polymers 0.000 description 4
- 239000005017 polysaccharide Substances 0.000 description 4
- 238000010345 tape casting Methods 0.000 description 4
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical compound CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 description 3
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 3
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 241000186660 Lactobacillus Species 0.000 description 3
- 229930195725 Mannitol Natural products 0.000 description 3
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000009826 distribution Methods 0.000 description 3
- 229940039696 lactobacillus Drugs 0.000 description 3
- 239000000594 mannitol Substances 0.000 description 3
- 235000010355 mannitol Nutrition 0.000 description 3
- 238000002390 rotary evaporation Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- JADVWWSKYZXRGX-UHFFFAOYSA-M thioflavine T Chemical compound [Cl-].C1=CC(N(C)C)=CC=C1C1=[N+](C)C2=CC=C(C)C=C2S1 JADVWWSKYZXRGX-UHFFFAOYSA-M 0.000 description 3
- 238000000492 total internal reflection fluorescence microscopy Methods 0.000 description 3
- 241001474374 Blennius Species 0.000 description 2
- 241000194033 Enterococcus Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 235000013373 food additive Nutrition 0.000 description 2
- 239000002778 food additive Substances 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 238000003672 processing method Methods 0.000 description 2
- 230000035899 viability Effects 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
- 229920001817 Agar Polymers 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000193171 Clostridium butyricum Species 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 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
- 241000194036 Lactococcus Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 239000012496 blank sample Substances 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 235000012970 cakes Nutrition 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
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- 239000012895 dilution Substances 0.000 description 1
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- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 235000015220 hamburgers Nutrition 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000003094 microcapsule Substances 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000008621 organismal health Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 229920006280 packaging film Polymers 0.000 description 1
- 239000012785 packaging film Substances 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 235000013772 propylene glycol Nutrition 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
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- 239000012798 spherical particle Substances 0.000 description 1
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- 235000010447 xylitol Nutrition 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
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
- A21—BAKING; EDIBLE DOUGHS
- A21D—TREATMENT, e.g. PRESERVATION, OF FLOUR OR DOUGH, e.g. BY ADDITION OF MATERIALS; BAKING; BAKERY PRODUCTS; PRESERVATION THEREOF
- A21D13/00—Finished or partly finished bakery products
- A21D13/30—Filled, to be filled or stuffed products
- A21D13/36—Filled wafers
-
- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3526—Organic compounds containing nitrogen
-
- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3553—Organic compounds containing phosphorus
-
- 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
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3562—Sugars; Derivatives thereof
-
- 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
- A23L33/17—Amino acids, peptides or proteins
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/185—Vegetable proteins
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B65—CONVEYING; PACKING; STORING; HANDLING THIN OR FILAMENTARY MATERIAL
- B65D—CONTAINERS FOR STORAGE OR TRANSPORT OF ARTICLES OR MATERIALS, e.g. BAGS, BARRELS, BOTTLES, BOXES, CANS, CARTONS, CRATES, DRUMS, JARS, TANKS, HOPPERS, FORWARDING CONTAINERS; ACCESSORIES, CLOSURES, OR FITTINGS THEREFOR; PACKAGING ELEMENTS; PACKAGES
- B65D65/00—Wrappers or flexible covers; Packaging materials of special type or form
- B65D65/38—Packaging materials of special type or form
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W90/00—Enabling technologies or technologies with a potential or indirect contribution to greenhouse gas [GHG] emissions mitigation
- Y02W90/10—Bio-packaging, e.g. packing containers made from renewable resources or bio-plastics
Abstract
The invention discloses a functional edible film carrying probiotics and a preparation method thereof, wherein the edible film takes a biopolymer as a matrix and is filled with probiotics liposome protected by protamine aggregate. The active probiotics are wrapped by liposome, and the slow release function is provided for the active probiotics. The surface of the liposome is modified by beta-cholesterol-D-glucoside (beta-CG), and can cooperate with zwitterionic lipid to induce protamine to generate spherical fiber aggregate around the liposome through hydrogen bond, wherein the diameter of the protamine fiber aggregate is 10-30um. The protamine has higher thermal stability, and fiber aggregate formed by the protamine forms stable carrier protection around liposome, so that the thermal stability of the probiotic active substance is improved. Can overcome the defect that the edible film can be produced only by a wet film forming method due to the heat instability characteristic of probiotics.
Description
Technical Field
The invention relates to the field of food processing, in particular to a functional edible film carrying probiotics and a preparation method thereof.
Background
The edible film is an edible film with specific functions, which is formed by interaction between film forming agent molecules of an edible biopolymer and a food additive through a certain processing technology. The prepared film can be directly applied to food as a package or a covering material to prolong the shelf life of the food. With the improvement of food quality requirements and the enhancement of environmental awareness, the functional edible film is increasingly widely applied in daily life of people. For example, various active compounds such as antioxidants, antibacterial agents, micronutrients, natural pigments, flavors, bioactive peptides, bacterial extracellular secondary metabolites, synbiotics, probiotics, metazoans, and the like may be added to the food packaging film. The food additive not only can help the active ingredients to exert the efficacy, but also can play a role in prolonging the shelf life of the food.
Wherein probiotics are microorganisms which can maintain the micro-ecological balance of host organisms and are beneficial to the health of the organisms. The probiotics used for human at present mainly comprise lactobacillus, bifidobacterium, enterococcus, lactococcus, streptococcus thermophilus and the like. However, probiotics are susceptible to changes in pH, oxygen concentration, temperature, etc. during production, storage and passage through the gastrointestinal tract, which can reduce their viability. In particular, the edible film containing probiotics can only be produced by a film forming method by a low-temperature wet method due to the heat instability characteristic of the probiotics cells. It is therefore important how to maintain high activity and stability of probiotics.
Disclosure of Invention
Aiming at the instability problems such as easy inactivation of probiotics, the invention provides a functional edible film loaded with probiotics and a preparation method thereof, which can improve the stability, delivery and absorption efficiency and application value of active ingredients.
The invention also provides a protamine aggregate-protected probiotic liposome, which is a nanoparticle with good stability, high intracellular delivery efficiency and certain antibacterial capability.
The specific technical scheme is as follows:
the functional edible film loaded with probiotics comprises the following raw materials in percentage by mass: 25-30% of prolamin, 15-20% of algal polysaccharide, 1-3% of plasticizer, 0.1-0.5% of protamine aggregate-protected probiotic liposome and the balance of solvent.
The prolamin comprises at least one of zein, wheat prolamin, etc.;
the seaweed polysaccharide is one or more of agar, sodium alginate and chitosan;
the plasticizer adopts at least one of glycerol, propylene glycol, sorbitol, xylitol, mannitol, ethylene glycol, glucose, fructose and the like;
the protamine aggregate-protected probiotic liposome is a spherical aggregate-protected probiotic liposome formed by protamine on the surface of the liposome. The probiotics are added into a lipid film formed by lipoid, beta-cholesterol-D-glucoside and an antioxidant to obtain the probiotics liposome, and the protamine forms spherical aggregates on the surface of the liposome to protect the probiotics liposome.
beta-cholesterol-D-glucoside (beta-CG) modification is adopted to cooperate with zwitterionic lipid to induce protamine fibers to form spherical aggregates on the surface of the liposome, and the liposome and the protamine are connected through a hydrogen bond network.
The functional edible film carrying probiotics is prepared by a tape casting method and is prepared according to the following steps:
s1, adding alcohol soluble protein into an ethanol solution, and stirring in a hot water bath until the alcohol soluble protein is dissolved;
s2, adding algal polysaccharide into glacial acetic acid, and stirring in a hot water bath until the algal polysaccharide is dissolved;
s3, uniformly mixing the solutions, adding a plasticizer and a protamine aggregate-protected probiotic liposome, fully stirring, performing cross-linking reaction in a hot water bath for a period of time, and performing vacuum degassing and defoaming;
and S4, pouring the solution on a clean and dry polytetrafluoroethylene film forming plate, and carrying out heat treatment to fix the film to obtain the functional edible film carrying probiotics.
In the S1, the mass concentration of the prolamin-ethanol solution is 9% -13%; the temperature of the hot water bath is 40-50 ℃.
In the step S2, the mass concentration of the seaweed polysaccharide-acetic acid solution is 3% -5%; the temperature of the hot water bath is 40-50 ℃.
In the step S3, the crosslinking reaction condition is 50-55 ℃, and the reaction is carried out for 1-2h in a water bath;
in the step S4, the heat treatment condition is 90-130 ℃ and the treatment time is 10-20min.
The invention also provides a protamine aggregate-protected probiotic liposome, which is obtained by adding probiotics into a lipid film formed by lipoid, beta-cholesterol-D-glucoside and an antioxidant, wherein the protamine forms a spherical aggregate on the surface of the liposome to protect the probiotic liposome.
Wherein: the lipid is zwitterionic lipid, including one or more of soybean lecithin, egg yolk lecithin, 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC), and dimyristoyl phosphatidylcholine (DMPC). The antioxidant is any one of VE acetate and propyl gallate. The molar ratio of the lipid to the beta-CG is 3.5-4:1, the mass ratio of the lipid to the probiotics is 6-15:1, and the molar ratio of the lipid to the protamine is 1.3-2:1.
The particle size of the probiotics liposome protected by protamine aggregate is 10-30um, and the probiotics liposome is uniformly distributed.
The preparation of the protamine aggregate-protected probiotic liposome comprises the following steps:
(1) Weighing lipid, beta-cholesterol-D-glucoside (beta-CG) and antioxidant, dissolving in organic solvent, evaporating solvent in rotary evaporator to form lipid film, and drying under vacuum overnight;
(2) Suspending the probiotic culture in a phosphate buffer to obtain a bacterial solution;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form liposome suspension, and homogenizing under high pressure to obtain probiotic liposome suspension;
(4) And (3) dissolving the protamine powder in deionized water, uniformly mixing with the liposome suspension, stirring and incubating for a period of time, and thus obtaining the protamine aggregate-protected probiotic liposome emulsion.
The lipid of the (1) is a zwitterionic lipid, and comprises one or more of soybean lecithin, egg yolk lecithin, 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) and dimyristoyl phosphatidylcholine (DMPC). The solvent preferably comprises at least one of chloroform, methanol, ethanol; the antioxidant is any one of VE acetate and propyl gallate; the addition amount of the antioxidant is 0.5% -1% of the mass of the liposome; the molar ratio of the lipoid to the beta-CG is 3.5-4:1.
Preferably, in the step (1), the temperature of the reduced pressure rotary evaporation is 40-50 ℃, the time of the reduced pressure rotary evaporation is 30-60 min, and the rotating speed of the reduced pressure rotary evaporation is 50-150 rpm.
Preferably, the concentration of the probiotics in the phosphate buffer solution in the step (2) is 0.01-0.03 mg/mL; the probiotics are selected from one or more strains of Lactobacillus, bifidobacterium, enterococcus, escherichia coli, bacillus, clostridium butyricum and Saccharomyces;
the phosphate buffer pH was 7.5.
In the step (3), the mass ratio of the lipoid to the probiotics is 6-15:1; the parameter conditions of the high-pressure homogenizing treatment are as follows: homogenizing for 1-10 times under 300-800bar pressure.
The condition of stirring and incubation in the step (4) is that the incubation is carried out for 16-24h at room temperature.
The concentration of the protamine aqueous solution in the step (4) is 0.2-0.5 mg/ml, and the molar ratio of the lipoid to the protamine is 1.3-2:1.
The particle size distribution of the prepared protamine aggregate generated around the probiotic liposome is uniform and is only 10-30um, which is far smaller than other protein aggregate microcapsules (1-10 mm).
The protamine is an alkaline protein, has high nutrition and functionality, can reduce blood pressure, promote digestion and the like, has high antibacterial capacity and high thermal stability, still has activity after being heated for 1 hour at 210 ℃, and can meet the requirements of various film forming processing methods by referring to the heat-resistant stability of the protamine.
Advantageous effects
The invention integrates the probiotic liposome into the edible film, and combines the preparation process of the liposome with the preparation process of the edible film after improving the heat resistance of the probiotic liposome, thereby completely meeting the requirements of various film forming processing methods. The prepared edible film has good antibacterial, antiseptic and fresh-keeping effects, and can be conveniently clamped in hamburgers, biscuits, cakes and the like, so that the edible film is greatly convenient for the cooperation of probiotics and other foods.
The invention provides a liposome, which can cooperate with zwitterionic lipid to induce the formation of protamine spherical fiber aggregate after being modified by beta-cholesterol-D-glucoside (beta-CG), and at the moment, the liposome and the protamine are mainly under the action of a hydrogen bond network. The survival rate and the thermal stability of the live bacteria are obviously improved after the probiotics are packaged by the method.
Drawings
FIG. 1 is a schematic diagram of the mechanism by which protamine forms spherical fiber aggregates on the surface of probiotic liposomes;
FIG. 2 is a total internal reflection fluorescence microscopy image of protamine aggregate protected probiotic liposomes;
fig. 3 is a graph of the particle size distribution of the probiotic liposome of example 1.
FIG. 4 shows the comparison of the number of active bacteria.
Figure 5 is a comparison of probiotic survival.
Detailed Description
Example 1
The bifidobacterium-loaded functional edible film comprises the following raw materials in percentage by mass: 25% of zein, 15% of chitosan, 1% of glycerol, 0.1% of protamine aggregate-protected bifidobacterium liposome and the balance of solvent.
The functional edible film carrying the bifidobacteria is prepared by a tape casting method and is prepared by the following steps:
step 1, adding 10g of zein into 90g of ethanol solution, stirring in a hot water bath at 50 ℃ until the zein is dissolved, and preparing a zein-ethanol solution with the mass concentration of 10%;
step 2, adding 6g of chitosan into 114g of glacial acetic acid, stirring in a hot water bath at 50 ℃ until the chitosan is dissolved, and preparing a zein-ethanol solution with the mass concentration of 5%;
step 3, uniformly mixing the zein-ethanol solution and the chitosan-glacial acetic acid solution, adding 0.4g of glycerol and 0.04g of bifidobacterium liposome protected by protamine aggregate, fully stirring at 300r/min, performing crosslinking reaction for 1h in a hot water bath at 55 ℃, and performing vacuum degassing and defoaming;
and 4, pouring the solution on a clean and dry polytetrafluoroethylene film forming plate, and performing heat treatment at 120 ℃ for 10min to fix the film to obtain the functional edible film carrying bifidobacteria.
The synthesis method of the protamine aggregate-protected bifidobacterium liposome comprises the following steps:
(1) 15.16mg of soybean lecithin, 13.56mg of DMPC, 3.87mg of beta-CG and 0.36mg of VE acetate were weighed and dissolved in chloroform, then the solvent was removed by evaporation in a 40℃water bath rotary evaporator at a rotation speed of 150rpm for 30min, forming a lipid film, followed by drying under vacuum overnight;
(2) 3.59mg of the bifidobacterium culture was suspended in 140mL of phosphate buffer at pH 7.5 to give a bifidobacterium solution at a concentration of 0.026 mg/mL;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form liposome suspension, and homogenizing at a high pressure of 800bar for 5 times to obtain bifidobacterium liposome suspension;
(4) 6.62mg of protamine powder is dissolved in 20mL of deionized water to prepare a mixed solution with the concentration of 0.33mg/mL, and then the mixed solution is uniformly mixed with the liposome suspension, stirred and incubated for 20h to prepare the bifidobacterium liposome emulsion with the concentration of 0.27mg/mL of protamine aggregate protection.
Example 2
The functional edible film with escherichia coli comprises the following raw materials in percentage by mass: 28% of zein, 17% of chitosan, 2% of mannitol, 0.4% of protamine aggregate-protected escherichia coli liposome and the balance of solvent.
The functional edible film carrying the escherichia coli is prepared by a tape casting method and is prepared according to the following steps:
step 1, adding 2.1g of zein into 14g of ethanol solution, stirring in a hot water bath at 45 ℃ until the zein is dissolved, and preparing a zein-ethanol solution with the mass concentration of 13%;
step 2, adding 1.27g of chitosan into 24.13g of glacial acetic acid, and stirring in a hot water bath at 50 ℃ until the chitosan is dissolved to prepare a chitosan-glacial acetic acid solution with the mass concentration of 5%;
step 3, after uniformly mixing the solutions, adding 0.15g mannitol and 0.03g protamine aggregate-protected escherichia coli liposome, fully stirring at 300r/min, performing crosslinking reaction for 1.5h in a hot water bath at 50 ℃, and performing vacuum degassing and defoaming;
and 4, pouring the solution on a clean and dry polytetrafluoroethylene film forming plate, and performing heat treatment at 100 ℃ for 20min to fix the film to obtain the functional edible film carrying bifidobacteria.
The liposome is an escherichia coli liposome protected by protamine aggregate, and the synthesis method comprises the following steps:
(1) 4.71mg of egg yolk lecithin, 19mg of POPC, 3.87mg of beta-CG and 0.15mg of VE acetate are weighed and dissolved in chloroform, and the solvent is evaporated in a water bath rotary evaporator at 50 ℃ for 40min at a speed of 100rpm to form a lipid film, and then dried under vacuum overnight;
(2) 2.37mg of E.coli culture was suspended in 100mL of phosphate buffer at pH 7.5 to give a bifidobacterium solution at a concentration of 0.024 mg/mL;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form liposome suspension, and homogenizing at a high pressure of 800bar for 5 times to obtain escherichia coli liposome suspension;
(4) 8.28mg of protamine powder is dissolved in 20mL of deionized water to prepare a mixed solution with the concentration of 0.41mg/mL, and then the mixed solution is uniformly mixed with the liposome suspension, stirred and incubated for 24 hours to prepare the escherichia coli liposome emulsion with the concentration of 0.32mg/mL of protamine aggregate protection.
Comparative example 1
The difference from example 1 is that: the bifidobacterium liposome is not added with beta-CG.
The specific synthesis method of the bifidobacterium liposome comprises the following steps:
(1) 15.16mg of soybean lecithin, 13.56mg of DMPC and 0.36mg of VE acetate were weighed out and dissolved in chloroform, after which the solvent was removed by evaporation in a 40℃water bath rotary evaporator at a rotation speed of 150rpm for 30min to form a lipid film, followed by drying under vacuum overnight.
(2) 3.59mg of the bifidobacterium culture was suspended in 140mL of phosphate buffer pH 7.5 to give a bifidobacterium solution at a concentration of 0.026 mg/mL;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form liposome suspension, and homogenizing at a high pressure of 800bar for 5 times to obtain bifidobacterium liposome suspension;
(4) 6.62mg of protamine powder is dissolved in 20mL of deionized water to prepare a mixed solution with the concentration of 0.33mg/mL, and then the mixed solution is uniformly mixed with the liposome suspension, stirred and incubated for 20h to prepare the bifidobacterium liposome emulsion with the concentration of 0.27 mg/mL.
Comparative example 2
The difference from example 1 is that: the bifidobacterium liposomes are not protected by protamine aggregates.
The specific synthesis method of the bifidobacterium liposome comprises the following steps:
(1) 15.16mg of soybean lecithin, 13.56mg of DMPC, 3.87mg of beta-CG and 0.36mg of VE acetate were weighed out and dissolved in chloroform, after which the solvent was removed by evaporation in a 40℃water bath rotary evaporator at a rotation speed of 150rpm for 30min, forming a lipid film, followed by drying under vacuum overnight.
(2) 3.59mg of the bifidobacterium culture was suspended in 140mL of phosphate buffer pH 7.5 to give a bifidobacterium solution at a concentration of 0.026 mg/mL;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form a liposome suspension, and homogenizing for 5 times under a high pressure of 800 bar.
The remainder were identical. Preparing the bifidobacterium liposome emulsion.
Comparative example 3
The difference from example 1 is that: the bifidobacterium liposome is not protected by protamine aggregate, and the film fixing heat treatment temperature is 50 ℃.
The bifidobacterium-loaded functional edible film comprises the following raw materials in percentage by mass: 25% of zein, 15% of chitosan, 1% of glycerol, 0.1% of bifidobacterium liposome and the balance of solvent.
The functional edible film carrying the bifidobacteria is prepared by a tape casting method and is prepared by the following steps:
step 1, adding 10g of zein into 90g of ethanol solution, stirring in a hot water bath at 50 ℃ until the zein is dissolved, and preparing a zein-ethanol solution with the mass concentration of 10%;
step 2, adding 6g of chitosan into 114g of glacial acetic acid, stirring in a hot water bath at 50 ℃ until the chitosan is dissolved, and preparing a zein-ethanol solution with the mass concentration of 5%;
step 3, uniformly mixing the zein-ethanol solution and the chitosan-glacial acetic acid solution, adding 0.4g of glycerol and 0.04g of bifidobacterium liposome protected by protamine aggregate, fully stirring at 300r/min, performing crosslinking reaction for 1h in a hot water bath at 55 ℃, and performing vacuum degassing and defoaming;
and 4, pouring the solution on a clean and dry polytetrafluoroethylene film forming plate, and performing heat treatment at 50 ℃ for 10min to fix the film to obtain the functional edible film carrying bifidobacteria.
The invention provides a bifidobacterium liposome, and the synthesis method comprises the following steps:
(1) 15.16mg of soybean lecithin, 13.56mg of DMPC, 3.87mg of beta-CG and 0.36mg of VE acetate were weighed out and dissolved in chloroform, after which the solvent was removed by evaporation in a 40℃water bath rotary evaporator at a rotation speed of 150rpm for 30min, forming a lipid film, followed by drying under vacuum overnight.
(2) 3.59mg of the bifidobacterium culture was suspended in 140mL of phosphate buffer at pH 7.5 to give a bifidobacterium solution at a concentration of 0.026 mg/mL;
(3) Adding the solution obtained in the step (2) into the lipid film obtained in the step (1), hydrating and swirling to form liposome suspension, and homogenizing at a high pressure of 800bar for 5 times to obtain the bifidobacterium liposome emulsion.
Test procedure
1. Total Internal Reflection Fluorescence Microscopy (TIRFM)
To observe the behavior of protamine fiber forming spherical aggregates on the surface of liposome, the sample emulsion prepared in example 1 was mixed with an equal amount of thioflavin T phosphate buffer solution at a concentration of 5uM, thioflavin T was specifically bound to fiber aggregates, and a 14uL mixed sample was extracted using a total internal reflection fluorescence microscope to observe the fluorescence image of the aggregates
As a result, as shown in FIG. 2, it was found that the linear fibers were stained with thioflavin T and exhibited fluorescence, and that protamine fibers were collected around the liposome to form spherical particles having a size of about 20. Mu.m.
2. Particle size and zeta potential test
The probiotic liposome emulsions of example 1, example 2 and comparative example 1 were diluted 10 times with deionized water and measured with a nanoparticle analyzer and zeta potential analyzer.
The average particle size and zeta potential of the measured probiotic liposome are shown in the following table 1, the average particle size of the probiotic liposome prepared in the examples and the comparative example is about 100-130nm, the particle size distribution graph of the probiotic liposome of the example 1 is shown in fig. 3, and the absolute value of zeta potential of the example is larger than 20mV, which is obviously higher than that of the comparative example 1. This demonstrates that the addition of beta-CG can properly raise the absolute zeta potential of the liposome surface, thereby further avoiding aggregation between liposomes.
TABLE 1
2. Probiotic viability test
Taking functional edible film samples prepared in the example 1, the comparative example 1 and the comparative example 2, performing ultrasonic demulsification by using a methanol solvent to release bacterial cells, performing anaerobic culture at 39 ℃ for 48-72 hours by using a plate viable count method, and calculating the colony number on the plate.
As shown in FIG. 4, the number of live probiotics in example 1 was 6.4X10 8 cfu/mL, whereas the number of viable bacteria without β -CG added to the liposomes in control 1 and without protamine aggregates added to the probiotic liposomes in control 2 was substantially absent. The method shows that the damage effect of high-temperature solid film heat treatment on probiotics can be avoided after the probiotics liposome is protected by protamine aggregate. In contrast, in comparative examples 1 and 2, since β -CG and protamine were not added, respectively, protamine aggregates were not formed on the surface of the probiotic liposomes, and thus the activity of the probiotics was greatly affected by the high-temperature film-fixing heat treatment.
3. Simulated gastric fluid digestion test
The prepared functional edible film carrying probiotics is subjected to in vitro simulation. The method for detecting the viable count of the probiotics comprises the following steps: the national standard GB/T4789.35-2016 food safety national standard food microbiology detection of lactobacillus is adopted.
Simulated gastric fluid preparation: to 1L of deionized water, 2g of sodium chloride and 0.26g of pepsin were added, and the pH of the solution was adjusted to 1 to 2 with 1M hydrochloric acid to prepare simulated gastric fluid. The samples to be tested prepared in example 1 and comparative example 3 were added to simulated gastric fluid in an amount of 1g, and at the same time, a probiotic edible film without liposome coating was used as a blank sample control. And incubating in a water bath shaker at 37 ℃ and 180rpm under constant temperature shaking, taking out 1mL of simulated gastric fluid at 60min, 120min and 180min respectively, and measuring the survival rate of probiotics by using a plate colony counting method after gradient dilution.
As shown in fig. 5, after the liposome Bao Maiyi is adopted for bacteria and is aggregated and protected by protamine fibers, the survival rate of the probiotics in the stomach environment is greatly improved. At the end of gastric digestion (180 min), the survival rate of the probiotics of example 1 was 96%, the survival rate of the liposome probiotics not protected by protamine fiber aggregation in comparative example 3 was 32%, and the survival rate of the probiotics without liposome encapsulation and without protamine fiber aggregation protection was only 5%. The result shows that after the probiotics are wrapped by the liposome, the survival rate of the probiotics in the gastric juice environment can be improved, the survival rate of the probiotics in the probiotics liposome which is prepared by the invention and is protected by the protamine fiber aggregation can be further greatly improved, the sample prepared in the example 1 is subjected to high-temperature heat treatment and basically has no damage to the probiotics, and the protamine fiber aggregation provides a certain protection effect for the probiotics liposome.
Claims (4)
1. The functional edible film loaded with probiotics is characterized by comprising the following raw materials in percentage by mass: 25-30% of prolamin, 15-20% of chitosan, 1-3% of plasticizer, 0.1-0.5% of protamine aggregate-protected probiotic liposome and the balance of solvent;
the preparation method of the protamine aggregate-protected probiotic liposome comprises the following steps:
(1) Weighing lipoid, beta-cholesterol-D-glucoside and antioxidant, dissolving in an organic solvent, evaporating to remove the solvent to form a lipid film, and then drying under vacuum overnight;
(2) Suspending the probiotic culture in a phosphate buffer solution to obtain a bacterial solution;
(3) Adding the solution in the step (2) into the lipid film in the step (1), hydrating and swirling to form liposome suspension, and homogenizing under high pressure to obtain probiotic liposome suspension;
(4) Dissolving protamine powder in deionized water, uniformly mixing the solution with the probiotic liposome suspension obtained in the step (3), and stirring and incubating to prepare the probiotic liposome protected by protamine aggregate;
the lipid is a zwitterionic lipid; the antioxidant is any one of VE acetate and propyl gallate;
the antioxidant is 0.5-1% of the liposome; the molar ratio of the lipid to the beta-cholesterol-D-glucoside is 3.5-4:1, the mass ratio of the lipid to the probiotics is 6-15:1, and the molar ratio of the lipid to the protamine is 1.3-2:1;
the preparation method of the functional edible film loaded with probiotics comprises the following steps:
s1, adding alcohol soluble protein into an ethanol solution, and stirring in a hot water bath until the alcohol soluble protein is dissolved;
s2, adding chitosan into glacial acetic acid, and stirring in a hot water bath until the chitosan is dissolved;
s3, uniformly mixing the solutions, adding a plasticizer and a protamine aggregate-protected probiotic liposome, fully stirring, performing cross-linking reaction in a hot water bath for a period of time, and performing vacuum degassing and defoaming;
and S4, pouring the solution on a clean and dry polytetrafluoroethylene film forming plate, and carrying out heat treatment to fix the film to obtain the functional edible film carrying probiotics.
2. The probiotic-loaded functional edible film according to claim 1, characterized in that the parameter conditions of the high-pressure homogenization treatment in step (3) are: homogenizing for 1-10 times under 300-800bar pressure.
3. The probiotic loaded functional edible film according to claim 1, wherein the prolamin is at least one of zein, wheat prolamin.
4. The probiotic-loaded functional edible film according to claim 1, characterized in that the mass concentration of the prolamin-ethanol solution in S1 is 9-13%; the mass concentration of the chitosan-acetic acid solution in the S2 is 3-5%; the temperature of the hot water bath is 40-50 ℃; in the step S3, the crosslinking reaction condition is 50-55 ℃, and the reaction is carried out for 1-2h in a water bath; in the step S4, the heat treatment condition is 90-130 ℃ and the treatment time is 10-20min.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102964848A (en) * | 2012-12-21 | 2013-03-13 | 青岛海尔软件有限公司 | Protein/polysaccharide composite edible film and preparation method thereof |
| CN104164090A (en) * | 2014-05-30 | 2014-11-26 | 中国计量学院 | EGCG nano liposome edible film and preparation method thereof |
| CN110122564A (en) * | 2019-04-25 | 2019-08-16 | 华南理工大学 | A kind of probiotics liposome and preparation method thereof |
| CN112754996A (en) * | 2021-03-16 | 2021-05-07 | 江西省科学院生物资源研究所 | Protamine short peptide modified paclitaxel liposome and preparation method thereof |
| CN113892650A (en) * | 2020-06-18 | 2022-01-07 | 湖南农业大学 | Probiotic liposome and preparation method thereof |
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| WO2004043363A2 (en) * | 2002-11-06 | 2004-05-27 | Azaya Therapeutics, Inc. | Protein-stabilized liposomal formulations of pharmaceutical agents |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102964848A (en) * | 2012-12-21 | 2013-03-13 | 青岛海尔软件有限公司 | Protein/polysaccharide composite edible film and preparation method thereof |
| CN104164090A (en) * | 2014-05-30 | 2014-11-26 | 中国计量学院 | EGCG nano liposome edible film and preparation method thereof |
| CN110122564A (en) * | 2019-04-25 | 2019-08-16 | 华南理工大学 | A kind of probiotics liposome and preparation method thereof |
| CN113892650A (en) * | 2020-06-18 | 2022-01-07 | 湖南农业大学 | Probiotic liposome and preparation method thereof |
| CN112754996A (en) * | 2021-03-16 | 2021-05-07 | 江西省科学院生物资源研究所 | Protamine short peptide modified paclitaxel liposome and preparation method thereof |
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