CN113575961A - 茶多酚益生乳酸菌粉剂及其制备方法 - Google Patents
茶多酚益生乳酸菌粉剂及其制备方法 Download PDFInfo
- Publication number
- CN113575961A CN113575961A CN202110870020.4A CN202110870020A CN113575961A CN 113575961 A CN113575961 A CN 113575961A CN 202110870020 A CN202110870020 A CN 202110870020A CN 113575961 A CN113575961 A CN 113575961A
- Authority
- CN
- China
- Prior art keywords
- tea polyphenol
- freeze
- lactic acid
- acid bacteria
- protective agent
- 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.)
- Pending
Links
- 150000008442 polyphenolic compounds Chemical class 0.000 title claims abstract description 127
- 235000013824 polyphenols Nutrition 0.000 title claims abstract description 127
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 title claims abstract description 118
- 239000000843 powder Substances 0.000 title claims abstract description 101
- 239000006041 probiotic Substances 0.000 title claims abstract description 64
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 63
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 62
- 241000894006 Bacteria Species 0.000 title claims abstract description 61
- 239000004310 lactic acid Substances 0.000 title claims abstract description 59
- 235000014655 lactic acid Nutrition 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 137
- 238000004108 freeze drying Methods 0.000 claims abstract description 99
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 78
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 78
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 78
- 239000003223 protective agent Substances 0.000 claims abstract description 72
- 235000020183 skimmed milk Nutrition 0.000 claims abstract description 58
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims abstract description 46
- 229930006000 Sucrose Natural products 0.000 claims abstract description 46
- 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 claims abstract description 46
- 239000005720 sucrose Substances 0.000 claims abstract description 46
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims abstract description 46
- 241001122767 Theaceae Species 0.000 claims description 123
- 239000000243 solution Substances 0.000 claims description 87
- 230000001580 bacterial effect Effects 0.000 claims description 74
- 239000007788 liquid Substances 0.000 claims description 45
- 239000012528 membrane Substances 0.000 claims description 43
- 238000001914 filtration Methods 0.000 claims description 41
- 230000001954 sterilising effect Effects 0.000 claims description 31
- 238000002156 mixing Methods 0.000 claims description 28
- 239000001963 growth medium Substances 0.000 claims description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 20
- 239000010802 sludge Substances 0.000 claims description 18
- 238000009630 liquid culture Methods 0.000 claims description 17
- 230000001678 irradiating effect Effects 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- 239000002609 medium Substances 0.000 claims description 6
- 239000008223 sterile water Substances 0.000 claims description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 claims description 3
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 3
- 239000000284 extract Substances 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229940099596 manganese sulfate Drugs 0.000 claims description 3
- 239000011702 manganese sulphate Substances 0.000 claims description 3
- 235000007079 manganese sulphate Nutrition 0.000 claims description 3
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- 239000001632 sodium acetate Substances 0.000 claims description 3
- 235000017281 sodium acetate Nutrition 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 2
- 230000018044 dehydration Effects 0.000 claims description 2
- 238000006297 dehydration reaction Methods 0.000 claims description 2
- 229920000136 polysorbate Polymers 0.000 claims description 2
- 206010018910 Haemolysis Diseases 0.000 abstract description 9
- 230000008588 hemolysis Effects 0.000 abstract description 9
- 238000007254 oxidation reaction Methods 0.000 abstract description 9
- 230000003647 oxidation Effects 0.000 abstract description 8
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 7
- 244000269722 Thea sinensis Species 0.000 abstract 3
- 235000011187 glycerol Nutrition 0.000 description 41
- 239000000203 mixture Substances 0.000 description 25
- 241000186660 Lactobacillus Species 0.000 description 19
- 229940039696 lactobacillus Drugs 0.000 description 19
- 230000000052 comparative effect Effects 0.000 description 16
- 238000007710 freezing Methods 0.000 description 12
- 230000008014 freezing Effects 0.000 description 12
- 230000005764 inhibitory process Effects 0.000 description 10
- 239000000523 sample Substances 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 235000013305 food Nutrition 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 230000004083 survival effect Effects 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 230000000968 intestinal effect Effects 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 210000001035 gastrointestinal tract Anatomy 0.000 description 4
- 230000002949 hemolytic effect Effects 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 230000002195 synergetic effect Effects 0.000 description 3
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- 244000063299 Bacillus subtilis Species 0.000 description 2
- 235000014469 Bacillus subtilis Nutrition 0.000 description 2
- -1 DPPH free radical Chemical class 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 230000002587 anti-hemolytic effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 239000002577 cryoprotective agent Substances 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 238000009777 vacuum freeze-drying Methods 0.000 description 2
- VZSRBBMJRBPUNF-UHFFFAOYSA-N 2-(2,3-dihydro-1H-inden-2-ylamino)-N-[3-oxo-3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)propyl]pyrimidine-5-carboxamide Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C(=O)NCCC(N1CC2=C(CC1)NN=N2)=O VZSRBBMJRBPUNF-UHFFFAOYSA-N 0.000 description 1
- APLNAFMUEHKRLM-UHFFFAOYSA-N 2-[5-[2-(2,3-dihydro-1H-inden-2-ylamino)pyrimidin-5-yl]-1,3,4-oxadiazol-2-yl]-1-(3,4,6,7-tetrahydroimidazo[4,5-c]pyridin-5-yl)ethanone Chemical compound C1C(CC2=CC=CC=C12)NC1=NC=C(C=N1)C1=NN=C(O1)CC(=O)N1CC2=C(CC1)N=CN2 APLNAFMUEHKRLM-UHFFFAOYSA-N 0.000 description 1
- YLZOPXRUQYQQID-UHFFFAOYSA-N 3-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)-1-[4-[2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidin-5-yl]piperazin-1-yl]propan-1-one Chemical compound N1N=NC=2CN(CCC=21)CCC(=O)N1CCN(CC1)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F YLZOPXRUQYQQID-UHFFFAOYSA-N 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- AYFVYJQAPQTCCC-GBXIJSLDSA-N L-threonine Chemical compound C[C@@H](O)[C@H](N)C(O)=O AYFVYJQAPQTCCC-GBXIJSLDSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 229930003451 Vitamin B1 Natural products 0.000 description 1
- 229930003448 Vitamin K Natural products 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 210000000436 anus Anatomy 0.000 description 1
- 235000003704 aspartic acid Nutrition 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000002366 mineral element Substances 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000004798 organs belonging to the digestive system Anatomy 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 1
- SHUZOJHMOBOZST-UHFFFAOYSA-N phylloquinone Natural products CC(C)CCCCC(C)CCC(C)CCCC(=CCC1=C(C)C(=O)c2ccccc2C1=O)C SHUZOJHMOBOZST-UHFFFAOYSA-N 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 210000001187 pylorus Anatomy 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 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
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/035—Organic compounds containing oxygen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
- A23L29/045—Organic compounds containing nitrogen as heteroatom
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- 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
-
- 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
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Health & Medical Sciences (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Animal Husbandry (AREA)
- Microbiology (AREA)
- Nutrition Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Physiology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明公开了一种茶多酚益生乳酸菌粉剂,属于生物技术领域,所述的粉剂包括菌种,所述的菌种为植物乳杆菌,且所述粉剂还包括冻干保护剂以及茶多酚,所述冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸。本发明还提供该粉剂的制备方法,通过本发明制备获得的茶多酚益生乳酸菌粉剂的抗氧化性、抑菌性性能以及抗溶血性都比较好具有较高的商业价值与推广价值。
Description
技术领域
本发明涉及生物技术领域,更具体地说,涉及一种茶多酚益生乳酸菌粉剂及其制备方法。
背景技术
肠道是人体重要的消化器官。肠指的是从胃幽门至肛门的消化管,是消化管中最长的一段,也是功能最重要的一段。肠道菌群,人体肠道中的正常微生物,如双歧杆菌、乳酸杆菌等能合成多种人体生长发育必须的维生素,如B族维生素(维生素B1、B2、B6、B12),维生素K、烟酸、泛酸等,还能利用蛋白质残渣合成必需氨基酸,如天冬门氨酸、苯丙氨酸、缬氨酸和苏氨酸等,并参与糖类和蛋白质的代谢,同时还能促进铁、镁、锌等矿物元素的吸收。这些营养物质对人类的健康有着重要作用,一旦缺少会引起多种疾病。
目前市面上存在着大量的益生菌粉剂,但一般的粉剂往往抑菌性能较差,起不到对肠道菌群调节的功能,且抗氧化性、抑菌性性能、抗溶血性也比较一般。
发明内容
本发明所要解决的技术问题是:提供一种茶多酚益生乳酸菌粉剂,以解决目前常规乳酸菌粉剂抗氧化性、抑菌性以及抗溶血活性性能一般的问题。
本发明所采取的技术方案是:提供一种茶多酚益生乳酸菌粉剂,所述的粉剂包括菌种,所述的菌种为植物乳杆菌,且所述粉剂还包括冻干保护剂以及茶多酚,所述冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸。
采用以上配方后,获得的茶多酚益生乳酸菌粉剂抗氧化性、抑菌性以及溶血活性都较好,DPPH自由基清除率较高,能够有效地对人体肠道菌群进行调节,并且本发明还首次将茶多酚引入至乳酸菌粉剂,进一步提高了粉剂的抑菌性。本发明制备获得的茶多酚益生乳酸菌粉剂在食品或畜牧业等行业的应用都极具前景。作为优选,所述植物乳杆菌、冻干保护剂以及茶多酚的质量配比为(1-2):(1-2):(0.001-0.002)。
作为优选,所述甘油、脱脂牛奶、蔗糖以及酪氨酸的质量配比为(2-4):(5-15):(5-15):(0.8-1.2)。
本发明另一方面提供上述茶多酚益生乳酸菌粉剂的制备方法,包括以下步骤:
S1、将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2、对甘油、脱脂牛奶、蔗糖以及酪氨酸的水溶液进行灭菌后混合得到冻干保护剂;
S3、在菌泥中加入冻干保护剂以及茶多酚溶液,冻干后得到茶多酚益生乳酸菌粉剂。
作为优选,所述步骤S1中,所述MARS液体培养基以质量配比计,包括以下组分:蛋白胨8-12份,牛肉膏粉4-6份,酵母浸粉3-5份,葡萄糖15-25份,磷酸氢二钾1-3份,乙酸钠4-6份,柠檬酸二铵1-3份,硫酸镁0.1-0.3份,硫酸锰0.03-0.07份,吐温801-2份。
作为优选,所述步骤S1中,所述培养的具体条件为:在37℃的温度条件下,将植物乳杆菌放置于MARS液体培养基中培养24h。
作为优选,所述步骤S2中,灭菌的具体条件为:将甘油放置在高压蒸汽灭菌锅中在121℃下灭菌20min,将脱脂牛奶在65℃恒温水浴锅中恒温30min,将蔗糖通过0.22μm滤膜过滤,将酪氨酸通过0.22μm滤膜过滤后继续通过紫外照射30min。
作为优选,所述步骤S3中,茶多酚溶液的浓度为0.1%,且茶多酚溶液配置后通过0.22μm滤膜过滤。
作为优选,所述步骤S1中,离心脱水的具体步骤为:将植物乳杆菌菌液放入冷冻离心机中以8000rpm/min的条件离心10min,之后使用无菌水洗涤2-3次后,得到菌泥。
作为优选,所述步骤S3中,冻干的条件为:将加入冻干保护剂以及茶多酚溶液后的菌泥放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h。
本发明通过预先将植物乳杆菌离心处理后,与本发明中制备获得的冻干保护剂以及茶多酚混合,本发明通过茶多酚、冻干保护剂以及植物乳杆菌的协同作用,进一步地提高了粉剂的抑菌性、抗溶血性以及抗氧化能力都十分地好,本发明的粉剂中,有效活菌大于1.85×108/g,且在制备的过程中对冻干保护剂进行了充分的除菌,最终制备获得的粉剂能够对肠道菌群进行有效地调节,因此将茶多酚益生乳酸菌粉剂的开发应用于市场,在食品或畜牧业等行业的应用都极具前景。
具体实施方式
下面将对本发明的技术方案进行清楚、完整地描述,显然,所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本发明实施例中选用的植物乳杆菌为宁波大学食品与药学院实验室中的植物乳杆菌L2,在今年的文章中进行过发布(Wu J,Yan X,Weng P,et al.Homology-and cross-resistance of Lactobacillus plantarum to acid and osmotic stress and theinfluence of induction conditions on its proliferation by RNA-Seq[J].Journalof Basic Microbiology,2021.),也可以使用其他常规的植物乳杆菌进行等同的替换。
实施例1
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比(质量配比,下同)的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:8:5:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%(质量配比,下同)的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
上述的MARS液体培养基包括以下组分:
成分 | 重量(g/L) |
蛋白胨 | 10.0 |
牛肉膏粉 | 5.0 |
酵母浸粉 | 4.0 |
葡萄糖 | 20.0 |
磷酸氢二钾 | 2.0 |
乙酸钠 | 5.0 |
柠檬酸二铵 | 2.0 |
硫酸镁 | 0.2 |
硫酸锰 | 0.05 |
吐温80 | 1.0 |
实施例2
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:10:10:1。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为10%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制10%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例3
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:12:15:1.2。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为12%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制15%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1.2%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例4
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为3:8:10:1.2。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制3%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制10%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1.2%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例5
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为3:10:15:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制3%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为10%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制15%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例6
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为3:12:5:1。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制3%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为12%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例7
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为4:8:15:1。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制4%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制15%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例8
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为4:10:5:1.2。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制4%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为10%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制1.2%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例9
一种茶多酚益生乳酸菌粉剂,由1:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为4:12:10:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制4%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为12%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制10%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例10:
一种茶多酚益生乳酸菌粉剂,由1:2:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:8:5:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:2:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例11:
一种茶多酚益生乳酸菌粉剂,由2:1:0.001配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:8:5:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以2:1:1的质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
实施例12:
一种茶多酚益生乳酸菌粉剂,由1:1:0.002配比的植物乳杆菌、冻干保护剂以及茶多酚制得,且所述的冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸,以质量配比计,甘油、脱脂牛奶、蔗糖以及酪氨酸的配比为2:8:5:0.8。
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制2%的甘油溶液,高压蒸汽灭菌锅中121℃下灭菌20min;用脱脂奶粉配制浓度为8%的脱脂牛奶,在65℃恒温水浴锅中恒温30min;配制5%的蔗糖溶液,并通过0.22μm滤膜过滤;配制0.8%的酪氨酸溶液,并通过0.22μm滤膜过滤后紫外照射30min;将等体积的以上溶液混合得到冻干保护剂;
S3配制0.2%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥、步骤S2制备的冻干保护剂以及茶多酚溶液以1:1:1的质量配比混合后,冻干,得到茶多酚益生乳酸菌粉剂。
对比例:
一种茶多酚益生乳酸菌粉剂,由1:0.001配比的植物乳杆菌、茶多酚制得,
所述茶多酚益生乳酸菌粉剂的制备方法包括以下步骤:
S1将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2配制0.1%的茶多酚溶液,并通过0.22μm滤膜过滤;将上述步骤S1制备获得的菌泥以及茶多酚溶液以及水溶液1:1:1质量配比混合后,放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h,得到茶多酚益生乳酸菌粉剂。
本发明实施例1-9以及对比例中步骤S1中采用的菌液为将植物乳杆菌统一放置在500mL的锥形瓶中进行培养,培养基为MARS液体培养基,在37℃下培养24h之后均分为10份获得,实施例2-9中采用的MARS液体培养基成分含量以及配比相同。
实施例1-9以及对比例的植物乳杆菌冻干存活率测定:
冻干前,将灭菌后的固体培养基倒入一次性培养皿中,等待培养基凝固,用100μl植物乳杆菌L2菌液,原始菌液浓度107CFU/mL,加入900MARS液体培养基稀释成10-1,按此方法将菌液稀释至-5、-6、-7三个梯度,每个固体培养基加入100稀释后的菌液,每个梯度做三个平板,放入37℃培养箱中培养24h,计算菌落数。冻干后使用同样方法,计算冻干前后的乳酸菌存活率。
实验初期将实施例1-10的植物乳杆菌统一放置在500mL的锥形瓶中进行培养,培养基为MARS液体培养基,在37℃下培养24h,将400mL菌液分装至10根离心管中,该10根离心管依次对应本发明上述的实施例1-9以及对比例,每管40mL,冻干后将冻干粉用无菌水复溶至40mL,均用涂布法计算菌落总数以及计算存活率。实验结果如下:
冻干前菌落总数
菌液稀释至10-5时菌落总数>300,10-7菌落总数<30,因此在10-6对菌落总数进行测定,植物乳杆菌L2总数为2.63×108。
冻干后菌落总数如下表所示:
组别 | 实施例 | 菌落总数(CFU) |
实施例1 | 1 | 2.32×10<sup>8</sup> |
实施例2 | 2 | 2.38×10<sup>8</sup> |
实施例3 | 3 | 2.27×10<sup>8</sup> |
实施例4 | 4 | 2.46×10<sup>8</sup> |
实施例5 | 5 | 2.62×10<sup>8</sup> |
实施例6 | 6 | 1.85×10<sup>8</sup> |
实施例7 | 7 | 2.67×10<sup>8</sup> |
实施例8 | 8 | 2.55×10<sup>8</sup> |
实施例9 | 9 | 1.99×10<sup>8</sup> |
对比例 | 10 | 1.78×10<sup>8</sup> |
由上表可知:实施例1-9以及对比例制备得到的茶多酚乳酸菌粉剂复溶后溶液乳酸菌均具有活性,说明真空冷冻干燥对于乳酸菌活性有保护作用;实施例1-9号添加冻干保护剂的组L2存活率均高于对照组,说明冻干保护剂对乳酸菌存活具有保护作用;实施例7存活率高于100%是因为茶多酚对于乳酸菌的生长具有促进作用,茶多酚对乳酸菌起到的成长促进作用也是同冻干保护剂协同作用产生的正效益。
实施例1-9以及对比例的抑菌性能测定:
本发明在平板计数时所使用的培养基为MARS固体培养基。其成分及含量如下表:
将配置好的固体培养基在121℃,20min条件下灭菌后待用,在超净台中倒平板,根据稀释涂布法计算菌落总数。
本发明在抑菌性测定中采用大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌作为测试的菌体,在培养大肠杆菌、金黄色葡萄球菌、枯草芽孢杆菌时,采用LB培养基,包括10g/L的胰蛋白胨、5g/L的酵母粉以及5g/L的氯化钠。
抑菌性采用牛津杯法测定,将冻干后的样品加入40mL无菌水复溶,每个在一次性培养皿中加入四个牛津杯,将灭菌后的固体培养基倒入一次性培养皿中,等待培养基凝固,将牛津杯取出,加入复溶后的样品,每个样品做两个牛津杯,放入37℃培养箱中培养24h,测量抑菌圈大小,抑菌直径<10mm说明无抑菌能力,抑菌圈直径>10mm说明有抑菌能力,抑菌圈直径>15mm说明抑菌能力强,测试的结果如下表所示:
由上表可知:实施例1-9以及对比例样品的抑菌圈直径均大于10mm,说明茶多酚乳酸菌粉剂均有抑菌作用;实施例1-9添加冻干保护剂的九组抑菌圈直径均大于对比例的抑菌圈直径,这也说明了冻干保护剂对乳酸菌和茶多酚的抑菌能力具有保护作用,同时冻干保护剂与茶多酚起到协同作用,进一步地加强了粉剂的抑菌性能;实施例7的抑菌圈直径最大,说明该实施例的抑菌效果相对较好。
实施例1-9以及对比例的溶血活性性能测定:
溶血活性采用血平板法测定。在血平板中加入100μl复溶后的样品,在血平板上均匀进行涂布,放入37℃培养箱中培养24h,观察是否有溶血现象,无溶血现象说明产品制备成功,经平板法测定后,发现实施例1-9均未出现溶血圈,而对比例每个菌落旁边都出现溶血圈,因此只有10号有溶血现象,说明添加冻干保护剂可以防止乳酸菌茶多酚样品出现溶血性。
实施例1-9以及对比例的抗氧化能力测定:
取实施例1-9以及对比例粉剂复溶后的样品液,配制成1.0mg/mL的样品液,以清除DPPH自由基的能力来测定抗氧化能力;用2mL0.2mol/L的DPPH甲醇溶液和2mL的上述样品溶液混合,在室温下进行避光反应30min,在517nm处测定吸光值Ai;空白组以等体积甲醇代替DPPH溶液,为A0;对比例以等体积蒸馏水代替样品溶液,
DPPH自由基的清除率按式下计算:
试验结果如下表所示:
组别 | DPPH自由基清除率% |
实施例1 | 78.10±0.70<sup>c</sup> |
实施例2 | 86.40±1.21<sup>d</sup> |
实施例3 | 74.27±0.72<sup>bc</sup> |
实施例4 | 90.53±1.22<sup>de</sup> |
实施例5 | 95.37±1.13<sup>ef</sup> |
实施例6 | 67.10±0.95<sup>a</sup> |
实施例7 | 97.47±1.01<sup>f</sup> |
实施例8 | 98.67±0.99<sup>f</sup> |
实施例9 | 72.37±1.27<sup>b</sup> |
对比例 | 66.50±0.53<sup>a</sup> |
如表3-3所示,实施例1-9以及对比例对于DPPH-自由基都有一定的清除率和抗氧化能力,1-9组对于自由基的清除率均明显高于对照组,这也说明了本发明加入的冻干保护剂起到了加强抗氧化性能的作用,其中实施例7清除率最高,说明实施例7的抗氧化能力相对较好。
本发明使用植物乳杆菌L2以及茶多酚作为原材料,通过真空冷冻干燥法制得粉剂,在冻干过程中添加甘油、脱脂牛奶、蔗糖以及酪氨酸四种原料组成的冻干保护剂,采用L9(34)正交试验方法设计的实施例1-9,通过测定冻干后粉剂的抗氧化性、抑菌性以及溶血活性,综合得出茶多酚-益生乳酸菌粉剂的较佳配比。按本发明配方制得的茶多酚乳酸菌粉剂无溶血现象,具有抑菌性,DPPH自由基清除率较高,最高达到了98.67%。并且茶多酚是茶叶中多酚类物质的总称,乳酸菌在传统上用于食品发酵,其菌体本身可以作为益生菌,对肠道菌群具有调节的功能,而粉剂又是现代食品行业广泛应用的食品形态,具有方便、成本低等特点。因此将茶多酚益生乳酸菌粉剂的开发应用于市场,在食品或畜牧业等行业的应用都极具前景。
以上就本发明较佳的实施例作了说明,但不能理解为是对权利要求的限制。本发明不仅局限于以上实施例,其具体结构允许有变化,凡在本发明独立要求的保护范围内所作的各种变化均在本发明的保护范围内。
Claims (10)
1.一种茶多酚益生乳酸菌粉剂,其特征在于,包括植物乳杆菌、冻干保护剂和茶多酚,所述冻干保护剂包括甘油、脱脂牛奶、蔗糖以及酪氨酸。
2.根据权利要求1所述的茶多酚益生乳酸菌粉剂,其特征在于,所述植物乳杆菌、冻干保护剂以及茶多酚的质量配比为(1-2):(1-2):(0.001-0.002)。
3.根据权利要求1所述的茶多酚益生乳酸菌粉剂,其特征在于,所述冻干保护剂中甘油、脱脂牛奶、蔗糖以及酪氨酸的质量配比为(2-4):(5-15):(5-15):(0.8-1.2)。
4.权利要求1项所述茶多酚益生乳酸菌粉剂的制备方法,其特征在于,包括以下步骤:
S1、将植物乳杆菌放置在MARS液体培养基下培养获得植物乳杆菌菌液,之后菌液离心脱水后得到植物乳杆菌菌泥;
S2、对甘油、脱脂牛奶、蔗糖以及酪氨酸的水溶液进行灭菌后混合得到冻干保护剂;
S3、在菌泥中加入冻干保护剂以及茶多酚溶液,冻干后得到茶多酚益生乳酸菌粉剂。
5.根据权利要求4所述的茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S1中,MARS液体培养基以质量配比计包括以下组分:蛋白胨8-12份,牛肉膏粉4-6份,酵母浸粉3-5份,葡萄糖15-25份,磷酸氢二钾1-3份,乙酸钠4-6份,柠檬酸二铵1-3份,硫酸镁0.1-0.3份,硫酸锰0.03-0.07份,吐温80 1-2份。
6.根据权利要求4所述的茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S1中,培养的具体条件为:在37℃的温度条件下,将植物乳杆菌放置于MARS液体培养基中培养24h。
7.根据权利要求4所述茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S2中,灭菌的具体条件为:将甘油放置在高压蒸汽灭菌锅中在121℃下灭菌20min,将脱脂牛奶在65℃恒温水浴锅中恒温30min,将蔗糖通过0.22μm滤膜过滤,将酪氨酸通过0.22μm滤膜过滤后继续通过紫外照射30min。
8.根据权利要求4所述茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S3中,茶多酚溶液的浓度为0.1%,且茶多酚溶液配置后通过0.22μm滤膜过滤。
9.根据权利要求4所述茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S1中,离心脱水的具体步骤为:将植物乳杆菌菌液放入冷冻离心机中以8000rpm/min的条件离心10min,之后使用无菌水洗涤2-3次后,得到菌泥。
10.根据权利要求4所述茶多酚益生乳酸菌粉剂的制备方法,其特征在于,所述步骤S3中,冻干的条件为:将加入冻干保护剂以及茶多酚溶液后的菌泥放入温度为-81℃的冰箱中冷冻2h后,继续加入至温度为-60℃的真空冷冻机中冷冻干燥36h。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110870020.4A CN113575961A (zh) | 2021-07-30 | 2021-07-30 | 茶多酚益生乳酸菌粉剂及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110870020.4A CN113575961A (zh) | 2021-07-30 | 2021-07-30 | 茶多酚益生乳酸菌粉剂及其制备方法 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113575961A true CN113575961A (zh) | 2021-11-02 |
Family
ID=78252545
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110870020.4A Pending CN113575961A (zh) | 2021-07-30 | 2021-07-30 | 茶多酚益生乳酸菌粉剂及其制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113575961A (zh) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20120196352A1 (en) * | 2009-07-22 | 2012-08-02 | Bong Joon Kim | Novel lactobacillus plantarum and composition comprising same |
US20120208260A1 (en) * | 2009-10-28 | 2012-08-16 | Cj Cheiljedang Corp. | Novel lactobacillus plantarum and composition comprising the same |
CN103918795A (zh) * | 2014-04-10 | 2014-07-16 | 陕西科技大学 | 一种茶多酚益生菌羊奶片及其制备方法 |
CN106880051A (zh) * | 2017-02-15 | 2017-06-23 | 黑龙江八农垦大学 | 一种植物乳杆菌软胶囊的制备方法 |
CN107495057A (zh) * | 2017-09-27 | 2017-12-22 | 广州市澳米环保科技有限公司 | 一种复合乳酸菌固体饮料及其制备方法 |
JP2020022392A (ja) * | 2018-08-07 | 2020-02-13 | 株式会社ヤクルト本社 | 乳酸菌凍結乾燥菌体の製造方法 |
CN111647510A (zh) * | 2020-06-18 | 2020-09-11 | 科兴生物制药股份有限公司 | 一种婴儿双歧杆菌冻干粉、制备方法及其所用复合保护剂 |
-
2021
- 2021-07-30 CN CN202110870020.4A patent/CN113575961A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20120196352A1 (en) * | 2009-07-22 | 2012-08-02 | Bong Joon Kim | Novel lactobacillus plantarum and composition comprising same |
US20120208260A1 (en) * | 2009-10-28 | 2012-08-16 | Cj Cheiljedang Corp. | Novel lactobacillus plantarum and composition comprising the same |
CN103918795A (zh) * | 2014-04-10 | 2014-07-16 | 陕西科技大学 | 一种茶多酚益生菌羊奶片及其制备方法 |
CN106880051A (zh) * | 2017-02-15 | 2017-06-23 | 黑龙江八农垦大学 | 一种植物乳杆菌软胶囊的制备方法 |
CN107495057A (zh) * | 2017-09-27 | 2017-12-22 | 广州市澳米环保科技有限公司 | 一种复合乳酸菌固体饮料及其制备方法 |
JP2020022392A (ja) * | 2018-08-07 | 2020-02-13 | 株式会社ヤクルト本社 | 乳酸菌凍結乾燥菌体の製造方法 |
CN111647510A (zh) * | 2020-06-18 | 2020-09-11 | 科兴生物制药股份有限公司 | 一种婴儿双歧杆菌冻干粉、制备方法及其所用复合保护剂 |
Non-Patent Citations (2)
Title |
---|
袁亚宏,岳田利,高振鹏,赵多勇: "冻干高活力乳酸菌粉保护剂的研究", 西北农林科技大学学报(自然科学版) * |
赵静;梁金钟;: "直投式产γ-氨基丁酸植物乳杆菌冻干保护剂生产工艺的优化", 食品工业科技 * |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
EP2647694B1 (en) | Dead lactobacillus biomass for antimicrobial use and a production method therefor | |
CN108102959B (zh) | 人源性降胆固醇植物乳杆菌zy08及其应用 | |
CN110218681B (zh) | 一株发酵乳杆菌kp101及其应用 | |
CN113088463B (zh) | 一种具有益生特性的嗜酸乳杆菌及其应用 | |
CN112812999B (zh) | 一株对阴沟肠杆菌具有抑制作用的植物乳杆菌slb01及其衍生产品和应用 | |
CN110495522B (zh) | 一种饲料用中药微生态制剂 | |
KR20210088408A (ko) | 락토바실러스 플란타럼 및 그의 용도 | |
CN112143680A (zh) | 具有抗氧化功效副干酪乳杆菌zjuids05及其应用 | |
KR102251295B1 (ko) | 락토바실러스 살리바리우스 v133 균주의 사균체를 유효성분으로 함유하는 숙취 예방 또는 해소용 조성물 | |
CN115927049A (zh) | 一株长双歧杆菌婴儿亚种b2-01及其应用 | |
CN111826324A (zh) | 一种鼠李糖乳杆菌菌粉的制备方法 | |
CN107828703B (zh) | 空间罗伊氏乳杆菌Fullarton-9-35及应用 | |
CN113088468B (zh) | 干酪乳杆菌Ma.GLRGJ 1及其应用 | |
CN112574924B (zh) | 一种枯草芽孢杆菌菌株及其微生态制剂和应用 | |
CN116814501B (zh) | 一种缓解肥胖的长双歧杆菌长亚种及其应用 | |
CN117448213A (zh) | 一种抑制产气荚膜梭菌的植物乳杆菌及其后生元和应用 | |
CN116875502A (zh) | 一种复合微生物制剂及应用 | |
KR20220004865A (ko) | 프로바이오틱스 관련 효소 분비능, 항산화 활성, 담즙산염 분해 활성, 항균 활성이 있고, 유해효소 및 유해대사산물을 생성하지 않는 락토바실러스 브레비스 srcm101607 균주 및 이의 용도 | |
CN110141584A (zh) | 一种开菲尔乳杆菌m11在抑菌及作为治疗ⅱ型糖尿病药剂的活性成分的应用 | |
CN113881592B (zh) | 一株罗伊氏乳杆菌及其应用 | |
CN115838661A (zh) | 一种植物乳杆菌扁鹊君18、植物乳杆菌制剂及其应用 | |
CN113575961A (zh) | 茶多酚益生乳酸菌粉剂及其制备方法 | |
RU2471864C1 (ru) | Пробиотический препарат против вирусных и бактериальных инфекций "токсиспорин", способ его получения, штамм бактерий bacillus licheniformis, используемый в качестве компонента пробиотического препарата | |
CN114806953A (zh) | 一种具有改善1型糖尿病特性的格氏乳杆菌 | |
CN114921383A (zh) | 一种具有清除胆固醇功能的益生菌制剂及其制备方法 |
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 |