CN115011517B - Emulsion, preparation method thereof and application of emulsion in probiotics preparation - Google Patents
Emulsion, preparation method thereof and application of emulsion in probiotics preparation Download PDFInfo
- Publication number
- CN115011517B CN115011517B CN202210661054.7A CN202210661054A CN115011517B CN 115011517 B CN115011517 B CN 115011517B CN 202210661054 A CN202210661054 A CN 202210661054A CN 115011517 B CN115011517 B CN 115011517B
- Authority
- CN
- China
- Prior art keywords
- oil
- emulsion
- preparation
- probiotics
- phase
- 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.)
- Active
Links
- 239000000839 emulsion Substances 0.000 title claims abstract description 99
- 239000006041 probiotic Substances 0.000 title claims abstract description 83
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 83
- 238000002360 preparation method Methods 0.000 title claims abstract description 56
- 230000000529 probiotic effect Effects 0.000 claims abstract description 44
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 38
- 239000002131 composite material Substances 0.000 claims abstract description 30
- 239000003995 emulsifying agent Substances 0.000 claims abstract description 20
- 239000003223 protective agent Substances 0.000 claims abstract description 15
- 239000012460 protein solution Substances 0.000 claims abstract description 6
- 241001052560 Thallis Species 0.000 claims abstract description 3
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 239000003921 oil Substances 0.000 claims description 38
- 235000019198 oils Nutrition 0.000 claims description 38
- 241000894006 Bacteria Species 0.000 claims description 37
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 36
- 150000001299 aldehydes Chemical class 0.000 claims description 29
- 238000003756 stirring Methods 0.000 claims description 22
- 108010046377 Whey Proteins Proteins 0.000 claims description 20
- 102000007544 Whey Proteins Human genes 0.000 claims description 20
- NEHNMFOYXAPHSD-UHFFFAOYSA-N citronellal Chemical compound O=CCC(C)CCC=C(C)C NEHNMFOYXAPHSD-UHFFFAOYSA-N 0.000 claims description 20
- 235000021119 whey protein Nutrition 0.000 claims description 20
- 239000012141 concentrate Substances 0.000 claims description 18
- 239000002304 perfume Substances 0.000 claims description 18
- KJPRLNWUNMBNBZ-QPJJXVBHSA-N (E)-cinnamaldehyde Chemical compound O=C\C=C\C1=CC=CC=C1 KJPRLNWUNMBNBZ-QPJJXVBHSA-N 0.000 claims description 12
- 229940117916 cinnamic aldehyde Drugs 0.000 claims description 12
- KJPRLNWUNMBNBZ-UHFFFAOYSA-N cinnamic aldehyde Natural products O=CC=CC1=CC=CC=C1 KJPRLNWUNMBNBZ-UHFFFAOYSA-N 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 12
- 238000010008 shearing Methods 0.000 claims description 12
- HGBOYTHUEUWSSQ-UHFFFAOYSA-N valeric aldehyde Natural products CCCCC=O HGBOYTHUEUWSSQ-UHFFFAOYSA-N 0.000 claims description 12
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 10
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 10
- 229930006000 Sucrose Natural products 0.000 claims description 10
- 229930003633 citronellal Natural products 0.000 claims description 10
- 235000000983 citronellal Nutrition 0.000 claims description 10
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 10
- 239000005720 sucrose Substances 0.000 claims description 10
- 239000003549 soybean oil Substances 0.000 claims description 8
- 235000012424 soybean oil Nutrition 0.000 claims description 8
- 230000035899 viability Effects 0.000 claims description 8
- ZTQSAGDEMFDKMZ-UHFFFAOYSA-N butyric aldehyde Natural products CCCC=O ZTQSAGDEMFDKMZ-UHFFFAOYSA-N 0.000 claims description 6
- -1 i.e. Substances 0.000 claims description 6
- NBBJYMSMWIIQGU-UHFFFAOYSA-N Propionic aldehyde Chemical compound CCC=O NBBJYMSMWIIQGU-UHFFFAOYSA-N 0.000 claims description 4
- 150000001720 carbohydrates Chemical class 0.000 claims description 4
- 239000008157 edible vegetable oil Substances 0.000 claims description 4
- JARKCYVAAOWBJS-UHFFFAOYSA-N hexanal Chemical compound CCCCCC=O JARKCYVAAOWBJS-UHFFFAOYSA-N 0.000 claims description 4
- 239000000341 volatile oil Substances 0.000 claims description 4
- 235000000346 sugar Nutrition 0.000 claims description 3
- WTEVQBCEXWBHNA-UHFFFAOYSA-N Citral Natural products CC(C)=CCCC(C)=CC=O WTEVQBCEXWBHNA-UHFFFAOYSA-N 0.000 claims description 2
- 229940043350 citral Drugs 0.000 claims description 2
- WTEVQBCEXWBHNA-JXMROGBWSA-N geranial Chemical compound CC(C)=CCC\C(C)=C\C=O WTEVQBCEXWBHNA-JXMROGBWSA-N 0.000 claims description 2
- 125000005456 glyceride group Chemical group 0.000 claims description 2
- FXHGMKSSBGDXIY-UHFFFAOYSA-N heptanal Chemical compound CCCCCCC=O FXHGMKSSBGDXIY-UHFFFAOYSA-N 0.000 claims description 2
- NUJGJRNETVAIRJ-UHFFFAOYSA-N octanal Chemical compound CCCCCCCC=O NUJGJRNETVAIRJ-UHFFFAOYSA-N 0.000 claims description 2
- 239000004006 olive oil Substances 0.000 claims description 2
- 235000008390 olive oil Nutrition 0.000 claims description 2
- 150000008163 sugars Chemical class 0.000 claims description 2
- MWOOGOJBHIARFG-UHFFFAOYSA-N vanillin Chemical compound COC1=CC(C=O)=CC=C1O MWOOGOJBHIARFG-UHFFFAOYSA-N 0.000 claims description 2
- FGQOOHJZONJGDT-UHFFFAOYSA-N vanillin Natural products COC1=CC(O)=CC(C=O)=C1 FGQOOHJZONJGDT-UHFFFAOYSA-N 0.000 claims description 2
- 235000012141 vanillin Nutrition 0.000 claims description 2
- 125000000185 sucrose group Chemical group 0.000 claims 1
- 239000003833 bile salt Substances 0.000 abstract description 7
- 230000004083 survival effect Effects 0.000 abstract description 6
- 210000004211 gastric acid Anatomy 0.000 abstract description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 3
- 230000036541 health Effects 0.000 abstract description 3
- 239000012528 membrane Substances 0.000 abstract description 3
- 210000001072 colon Anatomy 0.000 abstract description 2
- 238000009833 condensation Methods 0.000 abstract 1
- 230000005494 condensation Effects 0.000 abstract 1
- 125000002485 formyl group Chemical class [H]C(*)=O 0.000 abstract 1
- 239000003381 stabilizer Substances 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 43
- 230000029087 digestion Effects 0.000 description 26
- 239000008346 aqueous phase Substances 0.000 description 18
- 239000007788 liquid Substances 0.000 description 17
- 230000001580 bacterial effect Effects 0.000 description 16
- 230000000694 effects Effects 0.000 description 15
- 239000010802 sludge Substances 0.000 description 13
- 239000000203 mixture Substances 0.000 description 12
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 239000000725 suspension Substances 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 230000002411 adverse Effects 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 230000003213 activating effect Effects 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 238000009631 Broth culture Methods 0.000 description 6
- 239000004907 Macro-emulsion Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 230000001079 digestive effect Effects 0.000 description 6
- 239000012153 distilled water Substances 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 238000003760 magnetic stirring Methods 0.000 description 6
- 238000010907 mechanical stirring Methods 0.000 description 6
- 229920000223 polyglycerol Polymers 0.000 description 6
- WBHHMMIMDMUBKC-QJWNTBNXSA-M ricinoleate Chemical compound CCCCCC[C@@H](O)C\C=C/CCCCCCCC([O-])=O WBHHMMIMDMUBKC-QJWNTBNXSA-M 0.000 description 6
- 229940066675 ricinoleate Drugs 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 210000002784 stomach Anatomy 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 238000002474 experimental method Methods 0.000 description 5
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 210000000214 mouth Anatomy 0.000 description 4
- 230000001766 physiological effect Effects 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 210000000813 small intestine Anatomy 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 239000013589 supplement Substances 0.000 description 4
- 108010019160 Pancreatin Proteins 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 229940055695 pancreatin Drugs 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 238000013207 serial dilution Methods 0.000 description 3
- 238000004088 simulation Methods 0.000 description 3
- 235000013599 spices Nutrition 0.000 description 3
- 235000010469 Glycine max Nutrition 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 229940093761 bile salts Drugs 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000013068 control sample Substances 0.000 description 2
- 238000005538 encapsulation Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000000376 reactant Substances 0.000 description 2
- 230000009257 reactivity Effects 0.000 description 2
- 238000005057 refrigeration Methods 0.000 description 2
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 description 1
- 241000186016 Bifidobacterium bifidum Species 0.000 description 1
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 229920001661 Chitosan Polymers 0.000 description 1
- 238000000116 DAPI staining Methods 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 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
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 1
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 1
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 229920002494 Zein Polymers 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- FFBHFFJDDLITSX-UHFFFAOYSA-N benzyl N-[2-hydroxy-4-(3-oxomorpholin-4-yl)phenyl]carbamate Chemical compound OC1=C(NC(=O)OCC2=CC=CC=C2)C=CC(=C1)N1CCOCC1=O FFBHFFJDDLITSX-UHFFFAOYSA-N 0.000 description 1
- 229940002008 bifidobacterium bifidum Drugs 0.000 description 1
- 229940009289 bifidobacterium lactis Drugs 0.000 description 1
- 229940009291 bifidobacterium longum Drugs 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 235000011850 desserts Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000006353 environmental stress Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000002073 fluorescence micrograph Methods 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 235000013350 formula milk Nutrition 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229940071440 soy protein isolate Drugs 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 239000008307 w/o/w-emulsion Substances 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 239000005019 zein Substances 0.000 description 1
- 229940093612 zein Drugs 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/04—Preserving or maintaining viable microorganisms
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/23—Lactobacillus acidophilus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/46—Streptococcus ; Enterococcus; Lactococcus
-
- 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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicinal Preparation (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides an emulsion, a preparation method thereof and application thereof in probiotic emulsion, and belongs to the field of preparation of probiotic preparations. The emulsion is oil-in-water-in-oil W 1 /O/W 2 Composite emulsion structure. The emulsion is prepared from the following raw materials: a protective agent, a protein solution, an oil component, an oil-soluble emulsifier, an external water-soluble emulsifier and thalli. The invention encapsulates probiotics through a composite emulsion structure, and the stabilizer in the inner and outer water phases and the aldehyde in the oil phase modify W through aldehyde-amine condensation 1 O and O/W 2 The interface improves the stability of the interface membrane, further improves the entrapment and protection of probiotics, effectively reduces the damage of gastric acid and bile salt to the probiotics, improves the survival rate of the probiotics in the gastrointestinal tract, delivers more probiotics to the colon, and improves the health efficacy.
Description
Technical Field
The invention belongs to the field of preparation of probiotic preparations, and particularly relates to an emulsion, a preparation method thereof and application of the emulsion in probiotic emulsion.
Background
Probiotics are active microorganisms that have been shown to have health promoting effects on humans and animals, in particular positive physiological effects on promoting intestinal flora homeostasis in a host. Common probiotics are mainly bifidobacteria, lactobacilli (e.g. lactobacillus salivariusLactobacillus acidophilus and lactobacillus casei) and yeast. In recent years, the superior physiological function of probiotics has been attracting increasing research interest, including but not limited to maintaining intestinal microbial balance, stimulating the immune system and interfering with disease. To promote health, attempts have been made to supplement various food substrates (e.g., yogurt, dairy desserts, cheese, ice cream, and infant formulas) with probiotics. In order to function effectively, the probiotic bacterial body delivered to the target site-colon is generally in excess of 10 7 CFU/g or mL.
However, the disadvantage of probiotics being sensitive to environmental stress makes current probiotic supplement products not always able to reach this recommended level. Moreover, probiotic supplements have problems with processing, long-term storage and reduced probiotic viability in acidic conditions and digestive fluids (especially bile), which limits their use as food supplements. Thus, improving the stability of probiotics is an urgent challenge to be addressed by probiotic supplements in the food industry.
Disclosure of Invention
In order to solve the problems, the invention provides an emulsion, a preparation method thereof and application thereof in probiotic emulsion. The invention provides a W modified composite emulsion by perfume aldehyde 1/ O and O/W 2 The chemical reaction interface film is constructed at the interface, so that the stability and the encapsulation efficiency of the emulsion system are improved, and the influence of the external adverse environment on the probiotics in the content is isolated. The liquid preparation provided by the invention has the advantages that the storage stability is kept well, the oral digestion stability of probiotics in the storage period is obviously improved, the adverse effect of gastric acid environment on the probiotics can be obviously reduced, the survival rate of the probiotics in the digestion process is improved, and more probiotics can be delivered to the colon. Furthermore, the invention focuses on the change of the resistance of the probiotic liquid preparation to gastric acid, bile salts and other adverse environments in the storage stage, so that the probiotic liquid preparation can keep the activity of the probiotic in a longer period and better exert the physiological activity in the organism. Solves the problems of unstable probiotic-loaded composite emulsion system, poor acid and bile salt resistance effect and stability in storage processSex, etc.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an emulsion which is prepared from the following raw materials in percentage by mass: 3.5 to 11 percent of protective agent, 1 to 5 percent of protein solution, 25 to 40 percent of oil component, 2 to 5 percent of oil-soluble emulsifier, 2 to 5 percent of external water-soluble emulsifier and 8 to 10log CFU/mL of thalli calculated by the number of viable bacteria.
Preferably, the protective agent comprises: sugars and glycerol;
the saccharide includes: trehalose and/or sucrose and/or fructose and/or maltose and/or lactose and/or dextran; further preferred is sucrose;
the mass ratio of the saccharide to the glycerol is (2-20): (1-10), and more preferably 10:2.
in the present invention, the protein in the protein solution preferably includes: whey protein concentrate and/or soy protein isolate and/or zein and/or bovine serum albumin and/or soy peptide; further preferred is whey protein concentrate.
Preferably, the oil component consists of an edible oil and a flavor aldehyde essential oil, the edible oil comprising: soybean oil and/or medium chain glycerides and/or olive oil; the perfume aldehyde essential oil comprises: propionaldehyde and/or butyraldehyde and/or valeraldehyde and/or caproaldehyde and/or heptaldehyde and/or octaldehyde and/or citronellal and/or cinnamaldehyde and/or vanillin and/or citral.
Preferably, the external water phase water-soluble emulsifier comprises: whey protein concentrate and/or whey protein isolate and/or chitosan and/or casein and/or protein peptides.
The invention also provides a preparation method of the emulsion, which comprises the following steps:
(1) Preparing an inner water phase: mixing the thallus, the protective agent and the protein solution to obtain an inner water phase W1;
in the present invention, the internal water phase W 1 The mass percentage of the whey protein concentrate in the milk is preferably 2% -10%, and more preferably 4%;
the inner water phase W 1 The mass percentage of the saccharides in the protective agent is 2-20%, more preferably 10%, and the mass percentage of the glycerol is 1-10%, more preferably 2%;
(2) Preparing an oil phase: mixing the oil component and the oil-soluble emulsifier to obtain an oil phase O;
in the present invention, the content of perfume aldehyde in the oil component (relative to W 1 O) is 0 to 1wt%;
the content percentage of O in the oil phase (relative to W 1 50% -80% of O);
(3) Preparing a primary emulsion: inner water phase W 1 With the oil phase O to obtain a primary emulsion W by stirring and high-speed shearing 1 /O；
The oil phase O and the internal water phase W 1 The mass ratio of (2): 3, a step of;
(4) Preparing an emulsion: external water phase water-soluble emulsifier W 2 Phase and primary emulsion W 1 High-speed shearing of O to obtain W 1 /O/W 2 Composite emulsions, i.e., emulsions;
the external water phase water-soluble emulsifier W 2 With primary emulsion W 1 The mass ratio of/O was 1:1.
Preferably, the high-speed shearing rate in the step (3) is 12000r/min, and the high-speed shearing time is 10min.
Preferably, the high-speed shearing rate in the step (4) is 10000r/min, and the high-speed shearing time is 3min.
The invention provides application of the emulsion or the emulsion prepared by the preparation method in preparation of probiotic preparations.
The invention provides application of the emulsion or the emulsion prepared by the preparation method in preparation of a probiotic preparation for improving the survival activity of probiotics.
Preferably, the probiotics comprise: one or more of Bifidobacterium longum, bifidobacterium bifidum, streptococcus thermophilus, lactobacillus delbrueckii, lactobacillus bulgaricus, lactobacillus plantarum, bifidobacterium lactis, lactobacillus acidophilus, and Lactobacillus rhamnosus.
The emulsion provided by the invention can resist the effects of gastric acid and bile salts, so that the stability of the probiotic preparation prepared based on the emulsion in the oral digestion process is improved, and the probiotic preparation is beneficial to better exerting the physiological activity effect in organisms.
(1) Compared with the common composite emulsion preparation material and method, the preparation method provided by the invention has the advantages that the preparation process is simple, and compared with the common composite emulsion preparation material and method, a certain amount of perfume aldehyde is only required to be added during the preparation of the oil phase, and the protein in the water phase can be covalently crosslinked in the W during the formation of the composite emulsion preparation without additional preparation process 1 O and O/W 2 The interface generates a chemical interface film to improve stability and encapsulation efficiency and resist invasion of external bad environment.
(2) The protein added in the modified interface can react with perfume aldehyde and provide nutrition for life activities of probiotics in a storage period.
(3) The perfume aldehyde modified composite emulsion prepared by the invention has very stable and obvious protective effect on probiotics in the low-temperature storage process. The number of released viable bacteria after digestion of the fresh sample is higher than 6log CFU/mL, and after one month of storage, the number of released viable bacteria can reach 4log CFU/mL, which is far higher than that of a control sample in which the existence of viable bacteria can not be detected. The emulsion preparation modified by perfume aldehyde can protect probiotics from being damaged by gastric acid and bile salt to a greater extent, improve the survival rate of the probiotics, and can be used for developing novel probiotic foods, medicines and health care products.
Drawings
FIG. 1 is a confocal laser micrograph of a composite emulsion in which probiotics are embedded (the embedded bacteria are Lactobacillus plantarum and are stained with Hoechst 33342); the stable W/O/W emulsion is formed, the inner liquid drop presents obvious blue-green fluorescence, and the probiotics are proved to be encapsulated in the inner water phase and isolated from the external environment, so that the probiotics are protected;
FIG. 2 is a schematic representation of the oil droplets of a 10. Mu.L constant volume formed with a needle tube after 10800s of whey protein concentrate to extract a portion of the remaining oil droplets of the oil phase, the oil droplets being soy oil containing cinnamaldehyde; from the figure, the appearance of wrinkles and a distinct membrane structure at the interface is evident, indicating the presence of a solid interfacial membrane;
fig. 3 is a live/dead bacterial confocal fluorescence image of unencapsulated probiotic bacteria liquid (a), complex emulsion-entrapped probiotic bacteria (B), perfume aldehyde-modified complex emulsion-entrapped probiotic bacteria (C) after in vitro digestion, green on the way (DAPI staining) indicating live bacteria and red (PI staining) indicating dead bacteria.
Detailed Description
The following description refers to reagents or apparatus used without reference to specific techniques or conditions, and are carried out under conventional test conditions, and without reference to the specification of the reagent company, are carried out under conditions suggested by the specification. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention. The glassware, centrifuge tube, pipette tips, and suspensions and solutions used in each example were sterilized at 121℃for 15min.
Example 1
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: adding 4g oil soluble emulsifier polyglycerol ricinoleate (PGPR) into 55g soybean oil, stirring and heating at 65deg.C for 15min, adding 1g valeraldehyde, and magnetically stirring to obtain a uniform mixture
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: 2g of whey protein concentrate was added to 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 O) is added to 50g of the external aqueous phase W with magnetic stirring 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Example 2
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: adding 4g oil soluble emulsifier polyglycerol ricinoleate (PGPR) into 55g soybean oil, stirring and heating at 65deg.C for 15min, adding 1g citronellal, and magnetically stirring to obtain a uniform mixture
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: 2g of whey protein concentrate was added to 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 O) is added to 50g of the external aqueous phase W with magnetic stirring 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Example 3
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating probiotic lactobacillus plantarum with MRS broth,after 14h of culture in a constant temperature incubator at 37 ℃, the probiotic suspension is centrifuged at 4000rpm for 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: adding 4g oil soluble emulsifier polyglycerol ricinoleate (PGPR) into 55g soybean oil, stirring and heating at 65deg.C for 15min, adding 1g cinnamaldehyde, and magnetically stirring to obtain a uniform mixture
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: 2g of whey protein concentrate was added to 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 O) is added to 50g of the external aqueous phase W with magnetic stirring 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Example 4
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: 4g of oil-soluble emulsifier polyglycerol ricinoleate (PGPR) is added into 55g of soybean oil, stirred and heated for 15min at 65 ℃, and then 1g of compound aldehyde liquid (valeraldehyde, citronellal, cinnamaldehyde is 4:4:2) is added, and the mixture is stirred to be in a uniform state by a magnetic stirrer. The content of citronellal and valeraldehyde in the compound aldehyde liquid is the same.
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: 2g of whey protein concentrate was added to 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 O) is added to 50g of the external aqueous phase W with magnetic stirring 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Example 5
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: 4g of oil-soluble emulsifier polyglycerol ricinoleate (PGPR) is added into 55g of soybean oil, stirred and heated for 15min at 65 ℃, and then 1g of compound aldehyde liquid (valeraldehyde, citronellal, cinnamaldehyde is 3.5:3.5:3) is added, and the mixture is stirred by a magnetic stirrer until the mixture is uniformly mixed. The content of citronellal and valeraldehyde in the compound aldehyde liquid is the same.
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: adding 2g of whey protein concentrateTo 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 O) is added to 50g of the external aqueous phase W with magnetic stirring 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Comparative example 1
Activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial mud.
Comparative example 2
(1) Inner aqueous phase (W) 1 ) Is prepared from the following steps: activating the probiotic lactobacillus plantarum with MRS broth culture medium, culturing for 14 hours in a constant temperature incubator at 37 ℃, and centrifuging the probiotic suspension liquid under the condition of 4000rpm and 10min to obtain bacterial sludge. Uniformly mixing sterilized protective agent (2% glycerol, 10% sucrose, 4% whey protein concentrate) with the bacterial sludge to obtain an inner water phase.
(2) Preparation of an oil phase: 4g of polyglycerol ricinoleate (PGPR), an oil-soluble emulsifier, was added to 56g of soybean oil and stirred and heated at 65℃for 15min to dissolve completely.
(3) Primary emulsion W 1 Preparation of/O: 40g of internal aqueous phase W 1 To 60g of the oil phase, under mechanical stirring, at 1400rpm for 5min. The resulting macroemulsion was further homogenized under high shear at 12000rpm for 10min.
(4) External water phase (W) 2 ) Is prepared from the following steps: 2g of whey protein concentrate was added to 50g of sterile distilled water, stirred for two hours and left overnight at 4 ℃.
(5)W 1 /O/W 2 Preparation of the composite emulsion: 50g of the primary emulsion (W 1 In magnetic stirring barUnder-piece addition to 50g of external aqueous phase W 2 The crude emulsion was obtained at a stirring rate of 1000rpm for a stirring time of 5min. The crude emulsion obtained was further emulsified under high shear at 10000rpm for 3min.
Application example 1
Probiotic viability test
The invention provides a viability test experiment of a composite emulsion modified by perfume aldehyde during storage. The probiotic-loaded composite emulsions prepared in examples 1 to 5 and comparative example 2 were packaged and stored at 4 ℃. The viable count was measured by the dilution-coating plate method at the 0d, 7d, 14d and 28d, respectively, the composite emulsion was diluted with physiological saline, and the viable count was counted by fermentation under anaerobic conditions for 48 hours after coating on MRS solid medium. The test results are shown in Table 1.
TABLE 1 survival of probiotics during storage
* The superscripts A-D represent significant differences in the number of viable bacteria in the composite emulsion to which the different perfume aldehydes were added.
* Superscript a-d indicates the significant difference in the number of viable bacteria in the perfume aldehyde modified complex emulsion at different storage times.
The results in Table 1 demonstrate that the viable count of the samples stored in a refrigerator at 4℃for one month (except for example 3) remained on the same order of magnitude as the freshly prepared samples (10) 8 CFU/mL), combined with the reduced physiological activity of the probiotics reported in the study under low temperature refrigeration conditions, indicates that the survival rate of the probiotics is less affected by refrigeration at 4 ℃ for one month.
The activity of the reaction of the spice aldehyde and the protein and the stability of the product are related to the structure of the reactant, and three spice aldehydes (valeraldehyde, citronellal and cinnamaldehyde) which can be used as food additives are selected in the experiment according to the activity of the reaction, the stability of the system and the viability of bacteria so as to improve the capability of the composite emulsion for coating and protecting probiotics. The experimental results show that the sample of the experimental group (example 3) added with cinnamaldehyde has the lowest viable bacteria amount after 28d storage, and is reduced to 7.02log CFU/mL, which is related to the widely reported antibacterial performance. When the concentration of the cinnamaldehyde in the system is higher than the minimum antibacterial concentration, the killing effect of the cinnamaldehyde on probiotics is obvious. Secondly, the number of living bacteria of the experimental group samples (examples 1, 2, 4 and 5) except for the cinnamaldehyde is kept in the same order of magnitude as that of the control group after 28d storage, which shows that the influence of valeraldehyde, citronellal and low-concentration cinnamaldehyde (less than or equal to the minimum antibacterial concentration) on the viability of the probiotics is small, and the experimental group samples can be used for developing a probiotic entrapment and delivery system.
Application example 2
In vitro simulated digestion experiments
Oral digestion simulation: a simulated oral cavity simulated liquid was prepared with 0.5wt% mucin and various salts. In the oral simulated digestion experiments, 1.2mL of the complex emulsion was mixed with 7.8mL of phosphate buffer and 9mL of oral simulated liquid, and the final mixture contained 2wt% of oil. The pH of the mixture was adjusted to 6.8 and stirred at 37℃for 10min (100 r/min) to simulate the oral environment. After oral digestion was completed, 10-fold serial dilutions were performed with physiological saline, and 100 μl of the diluted solution was applied to MRS solid medium, fermented under anaerobic conditions for 48 hours, and the viable count was counted from the diluted solution of appropriate gradient.
Gastric digestion simulation: 1L of gastric simulated digest (SGF) was prepared by adding 2g of sodium chloride, 3.2g of pepsin and 7mL of hydrochloric acid to ultrapure water, with final sodium chloride, pepsin and hydrochloric acid concentrations of 2g/L,3.2g/L and 84mM, respectively. The SGF pH was adjusted to 1.5 and stored in an environment at 4deg.C for further use. Mixing oral cavity digestive juice and gastric simulated digestive juice collected from oral cavity simulated digestion experiment at a ratio of 1:1, adjusting pH to 2, and stirring in water bath at 37deg.C for 1 hr (100 r/min). After the completion of the stomach digestion, 10-fold serial dilutions were performed with physiological saline, and 100. Mu.L of the diluted solution with an appropriate gradient was applied to MRS solid medium, and the mixture was fermented under anaerobic conditions for 48 hours, followed by counting and measuring the number of viable bacteria.
Small intestine digestion simulation: simulated digestion of the small intestine is performedPreviously, 30mL of the digestive juice from the stomach was adjusted to pH 7.0, 3.5mL of bile salt extract was taken, and 1.5mL of CaCl was taken 2 Adding the mixed solution with NaCl into the digestive juice, stirring, adding 2.5 freshly prepared pancreatin suspension into the mixed solution, and adding 1g/L bile salt, 250mM NaCl,100mM CaCl into the final small intestine digestive juice 2 And 2g/L pancreatin. After pancreatin addition, the mixture was incubated at 37℃for 2h (100 r/min). After the completion of the stomach digestion, 10-fold serial dilutions were performed with physiological saline, and 100. Mu.L of the diluted solution with an appropriate gradient was applied to MRS solid medium, and the mixture was fermented under anaerobic conditions for 48 hours, followed by counting and measuring the number of viable bacteria. The test results are shown in tables 2 and 3.
TABLE 2 viability variation of probiotic bacteria entrapped in a composite emulsion formulation prepared from a single aldehyde during storage during digestion
* The superscript a-D indicates the significant difference in the number of viable bacteria in the composite emulsions with the addition of different perfume aldehydes.
* Superscript a-d indicates the significant difference in the number of viable bacteria in the perfume aldehyde modified complex emulsion at different storage times.
Table 2 data results are as above. First, the 0 day results indicate that the preparation of the composite emulsion results in a decrease in viable count in the system (from 9.21log CFU/mL to 8.73-8.35log CFU/mL) due to the disruption of probiotic activity by high shear during the preparation process. After simulated digestion in the oral cavity, stomach and small intestine, the viable count in the sample is reduced to 5.18-4.76log CFU/mL. The number of viable bacteria after digestion in the experimental group samples (examples 1 and 2), although higher than in control 1, was not significantly different from control 2, indicating that freshly prepared emulsions were not significantly effective in helping probiotics to resist adverse environmental effects.
Further, the results show that during 28d storage the probiotics' ability to resist the adverse environment changed significantly. Starting from 7d, the presence of viable bacteria was not detected after simulated digestion for comparative example 1 and comparative example 2, which, in combination with the viable bacteria count measured in table 1, demonstrates that although the probiotic viable bacteria count did not change significantly during low temperature storage, its resistance to adverse environments was significantly reduced. The samples (valeraldehyde and citronellal) of the experimental group can still detect the viable count higher than 2.92log CFU/mL at 7d and 14d, which shows that the effect of resisting the adverse environment is weak only by encapsulating the probiotics through the composite emulsion, and the effect of resisting the adverse environment by the interfacial film modified by the perfume aldehyde and the protein is obvious.
TABLE 3 variation of viability of probiotics entrapped by composite emulsions prepared from composite aldehydes in different proportions during storage during digestion
* The superscript a-D indicates the significant difference in the number of viable bacteria in the composite emulsions with the addition of different perfume aldehydes.
* Superscript a-d indicates the significant difference in the number of viable bacteria in the perfume aldehyde modified complex emulsion at different storage times.
Table 3 the data results are shown above. The reactivity and system stability of the aldehyde and protein reactions are related to the structure of the reactants, and the invention further optimizes the reaction by taking into account the reactivity, the characteristics of the product interfacial film, and the effect on the activity of the probiotics encapsulated in the system, with the option of adding multiple aldehydes in the emulsion preparation. The data result shows that the number of the detected viable bacteria after simulated digestion of the freshly prepared experimental group preparation (examples 4 and 5) is greater than 6.13log CFU/mL, compared with the number of the viable bacteria before digestion, the number of the viable bacteria is reduced by 2.17log CFU/mL, the number of the viable bacteria is obviously higher than that of the control example 2, the number of the viable bacteria is reduced by 3.95log CFU/mL, and the resistance effect on the gastrointestinal tract environment is better than that of the soft sweets reported in Chinese patent application No. 202111225493.5.
The storage stability of the emulsion was monitored. At 7d the control sample had no detectable presence of viable bacteria. The number of viable bacteria after simulated digestion is higher than 4.93log CFU/mL in the sample 7d of the experimental group, the number of viable bacteria is reduced along with the extension of storage time, and the maximum number of viable bacteria after digestion in the stomach and intestinal tracts can reach 3.95log CFU/mL in the sample 28 d. Furthermore, in combination with the data in Table 2, it was found that it is possible to adjust the aldehyde species to regulate the stability of the oil-water interface, and the effect of adding an emulsion of mixed aldehyde is more remarkable against the acidic environment of the stomach. Because the number of viable bacteria of the sample to which the mixed aldehyde is added after simulated digestion is significantly higher than that of a single kind of aldehyde in the same storage time. In conclusion, compared with the control example, the spice aldehyde modified composite emulsion prepared by the method under the simulated digestion condition has better protection effect on probiotics.
The foregoing is merely a preferred embodiment of the invention, and it should be noted that modifications and enhancements can be made to the inventors without departing from the principles of the present invention, which modifications and enhancements are also considered to be within the scope of the present invention.
Claims (1)
1. Use of an emulsion in the preparation of a probiotic preparation for increasing the viability of a probiotic;
the emulsion is prepared from the following raw materials in percentage by mass: 3.5 to 11 percent of protective agent, 1 to 5 percent of protein solution, 25 to 40 percent of oil component, 2 to 5 percent of oil-soluble emulsifier, 2 to 5 percent of external water-soluble emulsifier and 8 to 10log CFU/mL of thalli calculated by viable bacteria number;
the protective agent comprises: sugars and glycerol;
the saccharide is sucrose;
the mass ratio of the sugar to the glycerol is 10:2;
the oil component consists of edible oil and perfume aldehyde essential oil, wherein the edible oil comprises: soybean oil and/or medium chain glycerides and/or olive oil; the perfume aldehyde essential oil comprises: propionaldehyde and/or butyraldehyde and/or valeraldehyde and/or caproaldehyde and/or heptaldehyde and/or octaldehyde and/or citronellal and/or cinnamaldehyde and/or vanillin and/or citral;
the external water-soluble emulsifier is whey protein concentrate;
the preparation method of the emulsion comprises the following steps:
(1) Preparing an inner water phase: mixing the thallus, the protective agent and the protein solution to obtain an inner water phase W 1 ；
(2) Preparing an oil phase: mixing the oil component and the oil-soluble emulsifier to obtain an oil phase O;
(3) Preparing a primary emulsion: inner water phase W 1 With the oil phase O to obtain a primary emulsion W by stirring and high-speed shearing 1 /O；
The oil phase O and the internal water phase W 1 The mass ratio of (2): 3, a step of;
the high-speed shearing speed is 12000r/min, and the high-speed shearing time is 10min;
(4) Preparing an emulsion: external water phase water-soluble emulsifier W 2 Phase and primary emulsion W 1 High-speed shearing of O to obtain W 1 /O/W 2 Composite emulsions, i.e., emulsions;
the external water phase water-soluble emulsifier W 2 With primary emulsion W 1 The mass ratio of the component A to the component B is 1:1;
the high-speed shearing speed is 10000r/min, and the high-speed shearing time is 3min;
the probiotics are lactobacillus plantarum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210661054.7A CN115011517B (en) | 2022-06-13 | 2022-06-13 | Emulsion, preparation method thereof and application of emulsion in probiotics preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210661054.7A CN115011517B (en) | 2022-06-13 | 2022-06-13 | Emulsion, preparation method thereof and application of emulsion in probiotics preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115011517A CN115011517A (en) | 2022-09-06 |
| CN115011517B true CN115011517B (en) | 2024-02-06 |
Family
ID=83074038
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210661054.7A Active CN115011517B (en) | 2022-06-13 | 2022-06-13 | Emulsion, preparation method thereof and application of emulsion in probiotics preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115011517B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115558659B (en) * | 2022-10-28 | 2023-05-12 | 东北农业大学 | Lactobacillus plantarum embedding emulsion and preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112970929A (en) * | 2020-05-26 | 2021-06-18 | 华中农业大学 | Based on W1/O/W2Probiotic preparation with double-emulsion structure, preparation method and application |
| JP7061223B1 (en) * | 2020-10-30 | 2022-04-27 | 三栄源エフ・エフ・アイ株式会社 | Oil-in-water emulsification composition |
-
2022
- 2022-06-13 CN CN202210661054.7A patent/CN115011517B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112970929A (en) * | 2020-05-26 | 2021-06-18 | 华中农业大学 | Based on W1/O/W2Probiotic preparation with double-emulsion structure, preparation method and application |
| JP7061223B1 (en) * | 2020-10-30 | 2022-04-27 | 三栄源エフ・エフ・アイ株式会社 | Oil-in-water emulsification composition |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115011517A (en) | 2022-09-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111567669A (en) | Based on W1/O/W2Probiotic preparation with double-emulsion structure, preparation method and application | |
| CN101171019B (en) | Immune function modulating agents | |
| JP6483653B2 (en) | Survivability improver for lactic acid bacteria and / or bifidobacteria | |
| Marshall | Starter cultures for milk fermentation and their characteristics | |
| CN100469869C (en) | Embedding protection method for beneficial bacteria of intestinal tract | |
| ES2545209T3 (en) | Method of obtaining a new strain of Bifidobacterium bifidum with activity against Helicobacter pylori infection | |
| HUE025851T2 (en) | Mammalian milk microorganisms, compositions containing them and their use for the treatment of mastitis | |
| US20080063666A1 (en) | New lactic bacteria useful as probiotics | |
| JP6849972B2 (en) | Infection protection for babies | |
| CN115011517B (en) | Emulsion, preparation method thereof and application of emulsion in probiotics preparation | |
| WO2018190407A1 (en) | COMPOSITION FOR ACTIVATING Toll-LIKE RECEPTOR 2 | |
| JP6823302B2 (en) | Anti-allergic agent for babies | |
| RU2303058C2 (en) | Agent "biobalans-k" for treatment of intestine infections complicated with dysbacteriosis | |
| Percival | Choosing a probiotic supplement | |
| El Sayed et al. | Encapsulation of probiotics using mixed sodium alginate and rice flour to enhance their survivability in simulated gastric conditions and in UF-Kariesh cheese | |
| JP4489315B2 (en) | Method for producing fermented milk using egg yolk liquid containing anti-Helicobacter pylori antibody | |
| Gill et al. | Immune enhancement conferred by oral delivery of Lactobacillus rhamnosus HN001 in different milk-based substrates | |
| CN115137074A (en) | Defecation accelerant for infants | |
| Park et al. | Immunoenhancing effects of a new probiotic strain, Lactobacillus fermentum PL9005 | |
| RU2605626C2 (en) | Method of producing bacterial preparation with probiotic activity | |
| JP2003334065A (en) | Nucleic acid complex for protecting lactic acid bacterium | |
| EP2332557A1 (en) | Probiotic lactic acid bacteria | |
| Aspri | Donkey milk microbiota: isolation and characterization for potential applications | |
| O'Sullivan et al. | Bifidobacteria and siderophores produced thereby and methods of use | |
| CN116640689B (en) | Panda source Gao Liwei Shewanella kk64 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |