CN117801989B - Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis - Google Patents
Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis Download PDFInfo
- Publication number
- CN117801989B CN117801989B CN202311755723.8A CN202311755723A CN117801989B CN 117801989 B CN117801989 B CN 117801989B CN 202311755723 A CN202311755723 A CN 202311755723A CN 117801989 B CN117801989 B CN 117801989B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus
- acid
- parts
- strain
- preparation
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 60
- 239000002253 acid Substances 0.000 title claims abstract description 32
- 206010009900 Colitis ulcerative Diseases 0.000 title claims abstract description 30
- 201000006704 Ulcerative Colitis Diseases 0.000 title claims abstract description 30
- 239000003513 alkali Substances 0.000 title claims abstract description 26
- 240000006024 Lactobacillus plantarum Species 0.000 claims abstract description 51
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims abstract description 51
- 241000186604 Lactobacillus reuteri Species 0.000 claims abstract description 51
- 229940072205 lactobacillus plantarum Drugs 0.000 claims abstract description 51
- 229940001882 lactobacillus reuteri Drugs 0.000 claims abstract description 51
- 240000001046 Lactobacillus acidophilus Species 0.000 claims abstract description 50
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims abstract description 50
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims abstract description 50
- 239000003124 biologic agent Substances 0.000 claims abstract description 40
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 38
- 108010073771 Soybean Proteins Proteins 0.000 claims abstract description 37
- 241001134770 Bifidobacterium animalis Species 0.000 claims abstract description 36
- 229940118852 bifidobacterium animalis Drugs 0.000 claims abstract description 36
- 235000019710 soybean protein Nutrition 0.000 claims abstract description 35
- 239000003223 protective agent Substances 0.000 claims abstract description 31
- 239000001963 growth medium Substances 0.000 claims abstract description 25
- 238000000855 fermentation Methods 0.000 claims abstract description 17
- 230000004151 fermentation Effects 0.000 claims abstract description 17
- 238000013329 compounding Methods 0.000 claims abstract description 11
- 238000005507 spraying Methods 0.000 claims description 48
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 47
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 33
- 230000001580 bacterial effect Effects 0.000 claims description 29
- 241000894006 Bacteria Species 0.000 claims description 28
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 24
- URAYPUMNDPQOKB-UHFFFAOYSA-N triacetin Chemical compound CC(=O)OCC(OC(C)=O)COC(C)=O URAYPUMNDPQOKB-UHFFFAOYSA-N 0.000 claims description 22
- 229940086319 nattokinase Drugs 0.000 claims description 18
- 108010073682 nattokinase Proteins 0.000 claims description 18
- 239000000843 powder Substances 0.000 claims description 18
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 16
- 241000186000 Bifidobacterium Species 0.000 claims description 16
- 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 16
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 16
- 239000008103 glucose Substances 0.000 claims description 16
- 238000000034 method Methods 0.000 claims description 15
- 241001465754 Metazoa Species 0.000 claims description 14
- 150000002500 ions Chemical class 0.000 claims description 12
- 239000004925 Acrylic resin Substances 0.000 claims description 11
- 229920000858 Cyclodextrin Polymers 0.000 claims description 11
- 239000001856 Ethyl cellulose Substances 0.000 claims description 11
- ZZSNKZQZMQGXPY-UHFFFAOYSA-N Ethyl cellulose Chemical compound CCOCC1OC(OC)C(OCC)C(OCC)C1OC1C(O)C(O)C(OC)C(CO)O1 ZZSNKZQZMQGXPY-UHFFFAOYSA-N 0.000 claims description 11
- 239000001116 FEMA 4028 Substances 0.000 claims description 11
- 239000002202 Polyethylene glycol Substances 0.000 claims description 11
- WHGYBXFWUBPSRW-FOUAGVGXSA-N beta-cyclodextrin Chemical compound OC[C@H]([C@H]([C@@H]([C@H]1O)O)O[C@H]2O[C@@H]([C@@H](O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O[C@H]3O[C@H](CO)[C@H]([C@@H]([C@H]3O)O)O3)[C@H](O)[C@H]2O)CO)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H]3O[C@@H]1CO WHGYBXFWUBPSRW-FOUAGVGXSA-N 0.000 claims description 11
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims description 11
- 229960004853 betadex Drugs 0.000 claims description 11
- 229920001249 ethyl cellulose Polymers 0.000 claims description 11
- 235000019325 ethyl cellulose Nutrition 0.000 claims description 11
- 239000001087 glyceryl triacetate Substances 0.000 claims description 11
- 235000013773 glyceryl triacetate Nutrition 0.000 claims description 11
- 229920003132 hydroxypropyl methylcellulose phthalate Polymers 0.000 claims description 11
- 229940031704 hydroxypropyl methylcellulose phthalate Drugs 0.000 claims description 11
- 229920001223 polyethylene glycol Polymers 0.000 claims description 11
- 238000002791 soaking Methods 0.000 claims description 11
- 229960002622 triacetin Drugs 0.000 claims description 11
- 238000001816 cooling Methods 0.000 claims description 10
- 239000008213 purified water Substances 0.000 claims description 10
- 244000068988 Glycine max Species 0.000 claims description 9
- 235000010469 Glycine max Nutrition 0.000 claims description 9
- 235000004213 low-fat Nutrition 0.000 claims description 9
- 238000002156 mixing Methods 0.000 claims description 9
- 238000005406 washing Methods 0.000 claims description 9
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 claims description 8
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 claims description 8
- 229930003268 Vitamin C Natural products 0.000 claims description 8
- 229940024606 amino acid Drugs 0.000 claims description 8
- 150000001413 amino acids Chemical class 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 8
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 238000011081 inoculation Methods 0.000 claims description 8
- 230000004048 modification Effects 0.000 claims description 8
- 238000012986 modification Methods 0.000 claims description 8
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 8
- 238000004537 pulping Methods 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 229940073490 sodium glutamate Drugs 0.000 claims description 8
- 235000019154 vitamin C Nutrition 0.000 claims description 8
- 239000011718 vitamin C Substances 0.000 claims description 8
- 230000009286 beneficial effect Effects 0.000 claims description 4
- 230000001954 sterilising effect Effects 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 2
- 238000009928 pasteurization Methods 0.000 claims description 2
- 229940001941 soy protein Drugs 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims 1
- 230000037406 food intake Effects 0.000 claims 1
- 238000000265 homogenisation Methods 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 19
- 229960001231 choline Drugs 0.000 abstract description 14
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 abstract description 14
- 108010064851 Plant Proteins Proteins 0.000 abstract description 5
- 235000021118 plant-derived protein Nutrition 0.000 abstract description 5
- 210000002784 stomach Anatomy 0.000 abstract description 5
- 239000000825 pharmaceutical preparation Substances 0.000 abstract description 2
- 150000001450 anions Chemical class 0.000 description 14
- 210000004211 gastric acid Anatomy 0.000 description 12
- 230000000968 intestinal effect Effects 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 9
- 238000006243 chemical reaction Methods 0.000 description 8
- 210000001035 gastrointestinal tract Anatomy 0.000 description 8
- 238000005259 measurement Methods 0.000 description 7
- -1 oxygen ions Chemical class 0.000 description 7
- 206010059866 Drug resistance Diseases 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 239000003242 anti bacterial agent Substances 0.000 description 5
- 229940088710 antibiotic agent Drugs 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 239000006041 probiotic Substances 0.000 description 5
- 235000018291 probiotics Nutrition 0.000 description 5
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 4
- 239000002585 base Substances 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 206010012735 Diarrhoea Diseases 0.000 description 3
- YJQPYGGHQPGBLI-UHFFFAOYSA-N Novobiocin Natural products O1C(C)(C)C(OC)C(OC(N)=O)C(O)C1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-UHFFFAOYSA-N 0.000 description 3
- 230000000767 anti-ulcer Effects 0.000 description 3
- 210000000941 bile Anatomy 0.000 description 3
- 229910010293 ceramic material Inorganic materials 0.000 description 3
- 210000001072 colon Anatomy 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 229960002950 novobiocin Drugs 0.000 description 3
- YJQPYGGHQPGBLI-KGSXXDOSSA-N novobiocin Chemical compound O1C(C)(C)[C@H](OC)[C@@H](OC(N)=O)[C@@H](O)[C@@H]1OC1=CC=C(C(O)=C(NC(=O)C=2C=C(CC=C(C)C)C(O)=CC=2)C(=O)O2)C2=C1C YJQPYGGHQPGBLI-KGSXXDOSSA-N 0.000 description 3
- 150000007524 organic acids Chemical class 0.000 description 3
- 238000005245 sintering Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 2
- 241000186660 Lactobacillus Species 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000005868 electrolysis reaction Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 229940039696 lactobacillus Drugs 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 230000000644 propagated effect Effects 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 2
- WUBBRNOQWQTFEX-UHFFFAOYSA-N 4-aminosalicylic acid Chemical compound NC1=CC=C(C(O)=O)C(O)=C1 WUBBRNOQWQTFEX-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 239000005997 Calcium carbide Substances 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010022678 Intestinal infections Diseases 0.000 description 1
- 206010028140 Mucous stools Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 102000012479 Serine Proteases Human genes 0.000 description 1
- 206010041925 Staphylococcal infections Diseases 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 229960004909 aminosalicylic acid Drugs 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 235000021120 animal protein Nutrition 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229910000514 dolomite Inorganic materials 0.000 description 1
- 239000010459 dolomite Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000005684 electric field Effects 0.000 description 1
- 239000010411 electrocatalyst Substances 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 239000002662 enteric coated tablet Substances 0.000 description 1
- 230000003628 erosive effect Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 229910001385 heavy metal Inorganic materials 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000005027 intestinal barrier Anatomy 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical compound NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 description 1
- 229960004963 mesalazine Drugs 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 208000015688 methicillin-resistant staphylococcus aureus infectious disease Diseases 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- CLZWAWBPWVRRGI-UHFFFAOYSA-N tert-butyl 2-[2-[2-[2-[bis[2-[(2-methylpropan-2-yl)oxy]-2-oxoethyl]amino]-5-bromophenoxy]ethoxy]-4-methyl-n-[2-[(2-methylpropan-2-yl)oxy]-2-oxoethyl]anilino]acetate Chemical compound CC1=CC=C(N(CC(=O)OC(C)(C)C)CC(=O)OC(C)(C)C)C(OCCOC=2C(=CC=C(Br)C=2)N(CC(=O)OC(C)(C)C)CC(=O)OC(C)(C)C)=C1 CLZWAWBPWVRRGI-UHFFFAOYSA-N 0.000 description 1
- 239000002341 toxic gas Substances 0.000 description 1
- 231100000397 ulcer Toxicity 0.000 description 1
- 210000003556 vascular endothelial cell Anatomy 0.000 description 1
- 230000004580 weight loss Effects 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/43—Enzymes; Proenzymes; Derivatives thereof
- A61K38/46—Hydrolases (3)
- A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
- A61K38/482—Serine endopeptidases (3.4.21)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/10—Alcohols; Phenols; Salts thereof, e.g. glycerol; Polyethylene glycols [PEG]; Poloxamers; PEG/POE alkyl ethers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/38—Cellulose; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
- A61K47/40—Cyclodextrins; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/21—Serine endopeptidases (3.4.21)
- C12Y304/21062—Subtilisin (3.4.21.62)
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Inorganic Chemistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Tropical Medicine & Parasitology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Oncology (AREA)
- Communicable Diseases (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention provides a preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis, which belongs to the technical field of pharmaceutical preparations, and comprises the steps of adopting soybean protein to prepare a culture medium, performing micro-dynamic fermentation to culture bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri in an MRM molecular resonance environment, modifying each strain and compounding the strain; according to the invention, plant protein sources are treated through MRM molecular resonance and micro-dynamic fermentation culture is carried out in an MRM molecular resonance environment, so that the acid resistance and choline resistance of the amplified culture strain are improved; the strain is modified and protected by designing the protective agent and the film forming agent, the enteric film forming agent can protect the strain from being directly contacted with acid and alkali components in the stomach, the acid and alkali resistance of the strain is improved, the protective agent can protect the strain from being directly contacted with the film forming agent, and the strain is further protected to keep good activity.
Description
Technical Field
The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a preparation method of an acid and alkali resistant biological preparation for resisting ulcerative colitis.
Background
Ulcerative colitis is an inflammatory disease of the mucous membrane of the colon, which takes ulcer and erosion of the mucous membrane of the colon as main pathological changes and bloody mucous stool, abdominal pain and diarrhea as main clinical symptoms, and has the characteristics of slow disease course and repeated attacks. The existing treatment for ulcerative colitis mainly adopts antibiotics, such as mesalamine enteric-coated tablet and aminosalicylic acid preparation. However, the use of antibiotics can change the balance of the flora environment in the intestinal canal, and beneficial flora can be reduced by the damage of pharmaceutical ingredients, thereby causing damage to the immune system of the human body, affecting the self-repairing capability, having dependence and drug resistance on the antibiotics, and being harmful to the body functions after long-term administration of the antibiotics.
The common part of ulcerative colitis is a colon part of B type, belongs to an anaerobic part, is enriched with bifidobacterium anaerobic bacteria, and if antibiotics are used, the bifidobacterium anaerobic bacteria are reduced, so that harmful flora is propagated in a large amount, the flora environment is deteriorated, and diarrhea is caused.
The intestinal endogenous microorganism plays an important role in the treatment of ulcerative colitis, and the active substances secreted by the probiotics play an important role in regulating cytokine secretion, stabilizing intestinal barrier, enhancing immunity and the like. Therefore, the biological agent can improve the intestinal microecological environment, reach balance and further promote the recovery of ulcerative colitis.
After taking the biological agent, the biological agent can reach the intestinal environment of the affected part through the strict environment of strong gastric acid and strong choline to resist ulcerative colitis. However, probiotics are sensitive to living environment requirements, gastric acid resistance and choline resistance are generally weak, if the probiotics are contacted with strong gastric acid and choline, the probiotics can be inactivated, the activity is extremely low after the probiotics reach intestinal tracts, good flora environment improvement effect is difficult to be exerted, and the effect of resisting ulcerative colitis is poor. Therefore, there is a need to improve the acid and alkali resistance of biological agents to ensure biological activity.
Disclosure of Invention
Aiming at the problems of poor acid and alkali resistance, low activity and poor effect of improving the focus intestinal flora environment of the existing biological preparation. The invention provides a preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis, which comprises the steps of treating a vegetable protein source through MRM molecular resonance and performing micro-dynamic fermentation culture in an MRM molecular resonance environment, so that the acid resistance and choline resistance of an expanded culture strain are improved; the strain is modified and protected by designing the protective agent and the film forming agent, the enteric film forming agent can protect the strain from being directly contacted with acid and alkali components in the stomach, the acid and alkali resistance of the strain is improved, the protective agent can protect the strain from being directly contacted with the film forming agent, and the strain is further protected to keep good activity. The specific technical scheme is as follows:
a preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
S1, preparing soybean protein: washing and soaking organic low-fat soybeans by adopting MRM molecular resonance, and then pulping and curing, defoaming, sterilizing and cooling to obtain soybean proteins;
S2, preparing a culture medium: adding 90 g-110 g g soybean protein, 90 g-110 g g glucose, 1.8 g-2.2 g sodium bicarbonate, 1.8 g-2.2 g vitamin C, 2.8 g-3.2 g sodium glutamate, 0.25 g-0.35 g sodium chloride and 1.2 g-1.8 g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, and performing micro-dynamic fermentation culture under an MRM molecular resonance environment with the generation amount of negative ions of 900 e -/cm3 -1500 e -/cm3 respectively, so as to extract and obtain amplified cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the protectant comprises purified water, soy protein, and beta-cyclodextrin; the film forming agent comprises enteric polyacrylic resin, hydroxypropyl methylcellulose phthalate, ethyl cellulose, polyethylene glycol, glycerol and triacetin;
S5, bacteria compounding: and (3) compounding the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to obtain a biological preparation.
In the method S1, the soaking time is 6 h-10 h; the sterilization adopts a pasteurization method; the cooling temperature is 38-42 ℃.
The preparation method of the MRM molecular resonance water comprises the following steps: and (3) standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 900 e -/cm3 -1500 e -/cm3.
In the S2 of the method, 25-35% of brown sugar by mass percent is contained in the glucose, so that the strain can absorb amino acid conveniently.
In S3 of the above method, the sterile medium is homogenized after inoculation, and the homogenizing speed is 45rpm to 55rpm.
In the S3 of the method, the temperature of the micro-dynamic fermentation culture is 39-42 ℃ and the time is 5-7 h hours.
In the S4 of the method, the components of the protective agent comprise, by mass, 80-100 parts of purified water, 15-25 parts of soybean protein and 0.5-1 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100 (3-5).
In the step S4 of the above method, the film forming agent comprises the following components in parts by weight: 15 to 30 parts of enteric polyacrylic resin, 15 to 30 parts of hydroxypropyl methylcellulose phthalate, 15 to 30 parts of ethylcellulose, 2 to 5 parts of polyethylene glycol, 2 to 5 parts of glycerol and 2 to 5 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100 (3-5).
In the S5 of the method, the mass ratio of the animal bifidobacterium to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri= (35-55)/(20-40)/(5-15)/(10-15).
In the method S5, the biological agent is also added with nattokinase, and the added mass ratio is that the biological agent is nattokinase=100 (6-10).
Compared with the prior art, the preparation method of the acid and alkali resistant biological preparation for resisting ulcerative colitis has the beneficial effects that:
1. The invention adopts the organic low-fat soybean as the vegetable protein source, abandons bovine-derived animal protein from the source, realizes the efficient emulsification of the vegetable protein source through MRM molecular resonance water washing and soaking, does not need to add chemical components, and avoids the inhibition of chemical emulsifying agent on the activity of strains.
2. The preparation method of the invention utilizes MRM molecular resonance water to penetrate through the whole plant protein source preparation process and the strain culture process, adopts MRM molecular resonance to carry out physical displacement and sequencing on water molecules, activates the water molecules while the resonance effect, and jumps into small molecular groups from large molecular groups, thereby promoting the propagation of strains and the propagation speed of the strains to be improved by 1 time compared with that of the common culture method.
3. Surprisingly, it was found that by treating a vegetable protein source by MRM molecular resonance and performing a micro-dynamic fermentation culture in MRM molecular resonance environment, the acid resistance and choline resistance of the resulting strain are significantly enhanced. The inventor conjectures that MRM molecular resonance can improve the nutrition structure of a plant protein source, promote the absorption and utilization of the plant protein source, provide certain anions at the same time, and be beneficial to improving the immunity of the strain, so that the strain has stronger environment adaptability in micro-dynamic fermentation culture, and has higher acid resistance and choline resistance under the combined action of multiple aspects.
4. The anti-ulcerative colitis biological preparation has excellent acid resistance and choline resistance, can smoothly reach the intestinal tract through the stomach, is propagated in the intestinal tract, inhibits the growth of harmful bacteria, releases organic acid and nutrient substances as nutrients, further promotes the propagation of the anti-ulcerative colitis biological preparation, improves the microenvironment of the intestinal tract to reach balance, and achieves the purpose of resisting the ulcerative colitis.
5. The biological preparation adopts bifidobacterium animalis and lactobacillus plantarum as main treatment strains, can improve the immunity of human bodies, prevent diarrhea, reduce the side effects of antibiotic treatment, play a role in inhibiting harmful bacterial disorders in intestinal tracts, and utilize lactobacillus acidophilus and lactobacillus reuteri for assistance, effectively inhibit the growth of harmful bacteria, improve the distribution of intestinal flora, improve the environment, help the bifidobacterium animalis and lactobacillus plantarum to be delivered to target positions and have sufficient activity.
In addition, the bifidobacterium animalis also has the functions of resisting allergic intestinal infection and inflammation; lactobacillus plantarum also has the effect of inhibiting drug-resistant staphylococcus aureus MRSA; lactobacillus acidophilus is a strain which is regarded as third-generation lactobacillus fermentation in the lactobacillus family at present, and is an important microorganism in the intestinal tract of a human body; lactobacillus reuteri can antagonize the colonization of intestinal tracts by harmful bacteria, avoid suffering from intestinal tract diseases, and can also widely inhibit the growth of gram-positive bacteria, yeasts, fungi, protozoa and the like.
The combination of the interaction of four microbial flora can effectively fix in the intestinal canal, balance the microecology balance in the intestinal canal and form a protective barrier, activate damaged neutral flora, gradually repair focus by metabolic organic acid, and the organic acid does not generate stimulation stress reaction.
6. The temperature of the micro dynamic fermentation culture is 39-42 ℃, the culture temperature is higher than the conventional 37 ℃, and the propagation rate can be further improved under the condition of ensuring the activity.
7. The biological agent is also added with nattokinase, and the nattokinase is alkaline serine proteinase, has long half-life, strong specificity and small side effect, is directly taken, can promote vascular endothelial cells to secrete plasmin, and can promote recovery from formed fibrin or micro thrombus dissolution.
8. The invention designs the protective agent and the film forming agent to protect the bacterial strain, the enteric film forming agent can protect the bacterial strain from being directly contacted with acid and alkali components in the stomach, the acid and alkali resistance of the bacterial strain is improved, the protective agent can protect the bacterial strain from being directly contacted with the film forming agent, and the bacterial strain is further protected to keep good activity. The protective agent comprises soybean protein and beta-cyclodextrin, and is used for providing storage nutrition for soybean protein strains, keeping activity, and ensuring the coating compactness of the film forming agent due to the adhesion of the film forming agent; wherein the film forming agent comprises enteric polyacrylic resin, hydroxypropyl methylcellulose phthalate, ethylcellulose, polyethylene glycol, glycerol and triacetin; the components are matched, so that a good acid-base resistant film can be formed, the acid-base resistant film is not dissolved in the stomach, or the dissolution is delayed, the exposure time of the strain in acid-base is shortened, and the acid-base resistant film has good protection.
Detailed Description
The invention will be further illustrated with reference to specific examples, but the invention is not limited to these examples.
Example 1
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
s1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19% wt%) by adopting MRM molecular resonance for 8 h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 40 ℃ to obtain soybean protein;
the preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 1100 e -/cm3;
s2, preparing a culture medium: adding 100 g soybean protein, 100 g glucose, 2g sodium bicarbonate, 2g vitamin C, 3g sodium glutamate, 0.3 g sodium chloride and 1.5 g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
s3, culturing: inoculating bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 50rpm, then respectively performing micro-dynamic fermentation culture under the MRM molecular resonance environment with the generation amount of anions of 1100 e -/cm3, wherein the temperature is 40 ℃ and the time is 6h, and finally extracting to obtain the amplified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the components of the protective agent comprise 90 parts of purified water, 20 parts of soybean protein and 0.8 part of beta-cyclodextrin in parts by weight; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:4;
wherein, the film forming agent comprises the following components in parts by mass: 20 parts of enteric polyacrylic resin, 20 parts of hydroxypropyl methylcellulose phthalate, 20 parts of ethylcellulose, 3 parts of polyethylene glycol, 4 parts of glycerol and 3 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:4;
S5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=45:30:10:12, and the biological preparation is obtained.
According to the preparation method of the embodiment, glucose contains 30% of brown sugar by mass percent, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:8.
Example 2
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
s1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19 wt%) by adopting MRM molecular resonance for 6 h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 38 ℃ to obtain soybean protein;
The preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 900 e -/cm3;
S2, preparing a culture medium: adding 90g of soybean protein, 90g of glucose, 1.8g of sodium bicarbonate, 1.8g of vitamin C, 2.8g of sodium glutamate, 0.25g of sodium chloride and 1.2g of citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 45rpm, then respectively performing micro-dynamic fermentation culture under an MRM molecular resonance environment with the anion generation amount of 900 e -/cm3, wherein the temperature is 39 ℃ and the time is 5 hours, and finally extracting to obtain amplified and cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
wherein the components of the protective agent comprise, by mass, 80 parts of purified water, 15 parts of soybean protein and 0.5 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:3;
wherein, the film forming agent comprises the following components in parts by mass: 15 parts of enteric polyacrylic resin, 15 parts of hydroxypropyl methylcellulose phthalate, 30 parts of ethylcellulose, 2 parts of polyethylene glycol, 2 parts of glycerol and 2 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:3;
S5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=35:20:5:10, and the biological preparation is obtained.
According to the preparation method of the embodiment, 25 mass percent of brown sugar is contained in glucose, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:6.
Example 3
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
S1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19% wt%) by adopting MRM molecular resonance for 10 h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 42 ℃ to obtain soybean protein;
The preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 1500 e -/cm3;
s2, preparing a culture medium: adding 110 g soybean protein, 110 g glucose, 2.2 g sodium bicarbonate, 2.2 g vitamin C, 3.2 g sodium glutamate, 0.35 g sodium chloride and 1.8 g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 55rpm, then respectively performing micro-dynamic fermentation culture under the MRM molecular resonance environment with the anion generating amount of 1500 e -/cm3, wherein the temperature is 42 ℃ and the time is 7h, and finally extracting to obtain amplified cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the components of the protective agent comprise, by mass, 100 parts of purified water, 25 parts of soybean protein and 1 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:5;
wherein, the film forming agent comprises the following components in parts by mass: 30 parts of enteric polyacrylic resin, 20 parts of hydroxypropyl methylcellulose phthalate, 15 parts of ethylcellulose, 5 parts of polyethylene glycol, 5 parts of glycerol and 5 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:5;
S5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=55:40:15:15, and the biological preparation is obtained.
According to the preparation method of the embodiment, the glucose contains 35% of brown sugar by mass percent, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:10.
Example 4
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
S1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19% wt%) by adopting MRM molecular resonance for 6 h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 42 ℃ to obtain soybean protein;
The preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 900 e -/cm3;
S2, preparing a culture medium: adding 110 g soybean protein, 90g glucose, 2.2 g sodium bicarbonate, 1.8g vitamin C, 3.2 g sodium glutamate, 0.25g sodium chloride and 1.8g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 45rpm, then respectively performing micro-dynamic fermentation culture under the MRM molecular resonance environment with the anion generating amount of 1500 e -/cm3, wherein the temperature is 39 ℃ and the time is 7h, and finally extracting to obtain amplified cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the components of the protective agent comprise, by mass, 80 parts of purified water, 25 parts of soybean protein and 0.5 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:5;
wherein, the film forming agent comprises the following components in parts by mass: 15 parts of enteric polyacrylic resin, 30 parts of hydroxypropyl methylcellulose phthalate, 20 parts of ethylcellulose, 5 parts of polyethylene glycol, 2 parts of glycerol and 5 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:3;
S5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=55:20:15:10, and the biological preparation is obtained.
According to the preparation method of the embodiment, 25 mass percent of brown sugar is contained in glucose, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:10.
Example 5
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
s1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19% wt%) by adopting MRM molecular resonance for 10 h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 38 ℃ to obtain soybean protein;
The preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 1500 e -/cm3;
s2, preparing a culture medium: adding 90g of soybean protein, 110 g g of glucose, 1.8g of sodium bicarbonate, 2.2 g vitamin C, 2.8g of sodium glutamate, 0.35 g sodium chloride and 1.2g of citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 55rpm, then respectively performing micro-dynamic fermentation culture under an MRM molecular resonance environment with the anion generation amount of 900 e -/cm3, wherein the temperature is 42 ℃ and the time is 5 hours, and finally extracting to obtain amplified and cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the components of the protective agent comprise, by mass, 100 parts of purified water, 15 parts of soybean protein and 1 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:3;
Wherein, the film forming agent comprises the following components in parts by mass: 30 parts of enteric polyacrylic resin, 15 parts of hydroxypropyl methylcellulose phthalate, 20 parts of ethylcellulose, 2 parts of polyethylene glycol, 5 parts of glycerol and 2 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:5;
S5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=35:40:5:15, and the biological preparation is obtained.
According to the preparation method of the embodiment, the glucose contains 35% of brown sugar by mass percent, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:6.
Example 6
A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis comprises the following steps:
s1, preparing soybean protein: washing and soaking organic low-fat soybeans (fat content of 17 wt% -19 wt%) by adopting MRM molecular resonance for 7h, pulping and curing, defoaming, pasteurizing (90 ℃ -95 ℃), and cooling to 41 ℃ to obtain soybean protein;
The preparation method of the MRM molecular resonance water comprises the following steps: standing the MRM molecular resonance plate in water for reaction to form molecular resonance water with the anion generating amount of 1300 e -/cm3;
S2, preparing a culture medium: adding 95 g soybean protein, 105g glucose, 2.1g sodium bicarbonate, 1.9 g vitamin C, 3.1 g sodium glutamate, 0.32 g sodium chloride and 1.6g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, homogenizing after inoculation, wherein the homogenizing speed is 50rpm, then respectively performing micro-dynamic fermentation culture under the MRM molecular resonance environment with the anion generating amount of 1300 e -/cm3, wherein the temperature is 41 ℃ and the time is 6h, and finally extracting to obtain the amplified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the components of the protective agent comprise, by mass, 100 parts of purified water, 20 parts of soybean protein and 0.6 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100:3;
wherein, the film forming agent comprises the following components in parts by mass: 25 parts of enteric polyacrylic resin, 25 parts of hydroxypropyl methylcellulose phthalate, 15 parts of ethylcellulose, 4 parts of polyethylene glycol, 4 parts of glycerol and 3 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100:4;
s5, bacteria compounding: the modified bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are compounded, wherein the mass ratio of the bifidobacterium animalis to the lactobacillus plantarum to the lactobacillus acidophilus to the lactobacillus reuteri=50:35:12:14, and the biological preparation is obtained.
According to the preparation method of the embodiment, glucose contains 30% of brown sugar by mass percent, so that the strain can absorb amino acid conveniently. The biological agent is also added with nattokinase with the mass ratio of the biological agent to the nattokinase=100:8.
In the preparation methods of examples 1 to 6, the preparation method of the MRM molecular resonator plate specifically includes: the molecular resonance plate comprises a ceramic material of the floor tile type with an intermediate layer and a negative ion generating material wrapped around the ceramic material of the floor tile type. The ceramic material of the floor tile is formed by mixing 15 mass percent of molecular resonance powder into glaze and performing anaerobic sintering, wherein the anaerobic sintering temperature is 1200 ℃, and the anaerobic sintering time is 8 hours; wherein the formula of the molecular resonance powder comprises 40% of carbon, 30% of ceramic, 20% of dolomite and 10% of calcium carbide by mass. The negative ion generation amount of the negative ion generation material is 900-1500 e-/cm 3.
Negative ions generated by the molecular resonance material: the method is characterized in that oxygen ions with negative charges and redundant paired electrons are obtained, the technology is also called as a soft discharge technology, the generated negative ions are electrons with moderate energy by combining a discharge electrode with superconducting characteristics formed by adopting a technology similar to the pulse energy enhancement of the ions, so that high-concentration and high-quality pure negative ions (also called as electrocatalyst materials) are generated, the negative ions are nearly superconducting materials, the resistance is nearly zero, hydrogen ions generated by water electrolysis rapidly move to a negative electrode in an electric field generated by the negative electrode, one part of the hydrogen ions absorb electrons to become hydrogen, and the other part of the hydrogen ions are combined with water molecules to form active molecules, namely hydronium ions (H 3O+), so that the negative electrode has extremely strong emulsifying property; in addition, hydroxyl ions generated by water electrolysis form H 3O2 - anions, namely 'hydroxyl ions', with other water molecules. Therefore, a strong resonance effect can be generated, free precipitation of electric ions is utilized, no derivative exists, negative ions have extremely strong penetrability and dissolving power, toxic gas release is facilitated, heavy metals sink, and safety of plant protein sources is improved.
Performance testing
1. Determination of drug resistance
The bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri obtained by the expansion culture in the step S3 in the preparation method of the example 1 are taken for drug resistance measurement, and the results are shown in the table 1:
TABLE 1 determination of drug resistance (diameter cm of inhibition zone)
From the results, it was found that bifidobacterium animalis and lactobacillus plantarum have good resistance to chloramphenicol, ampicillin, kanamycin and novobiocin, lactobacillus acidophilus has good resistance to kanamycin and novobiocin, lactobacillus reuteri has excellent resistance to kanamycin, and lactobacillus reuteri has good resistance to novobiocin. The strain has better drug resistance.
The strain resistance obtained by the expansion culture in the step S3 in the preparation methods of the examples 2 to 6 is equivalent to that of the strain of the example 1.
2. Antibacterial ability measurement
The bacteriostasis ability of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri obtained by the expansion culture in the step S3 in the preparation method of the example 1 is measured, and the results are shown in Table 2:
TABLE 2 determination of antibacterial ability (diameter of inhibition zone cm)
From the above results, it was found that Lactobacillus plantarum, lactobacillus acidophilus and Lactobacillus reuteri have bacteriostatic effects on harmful bacteria.
The bacteria inhibition capacity obtained by the expansion culture of the step S3 in the preparation methods of the examples 2 to 6 is equivalent to that of the bacteria inhibition capacity of the example 1.
3. Gastric acid resistance assay
Taking bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri obtained by the expansion culture in the step S3 in the preparation method of the example 1 for gastric acid resistance measurement; and synchronously measuring gastric acid resistance of the finished biological preparation prepared in the example 1, and the results are shown in Table 3:
TABLE 3 acid resistance measurement results
Testing 3000 strains of single bacteria; mixed bacteria 12000 strains were tested, containing 3000 strains each.
From the above results, it is clear that the single strain itself has a general survival rate in simulated gastric acid at ph2.5 and has an excellent survival rate in simulated gastric acid at ph3.0 and ph 3.5; the survival rate of the modified mixed bacteria in simulated gastric acid at pH2.5, pH3.0 and pH3.5 is improved, and especially the survival rate can be greatly improved by more than 92% in simulated gastric acid at pH2.5 for 4 hours.
The gastric acid resistance of the strain obtained by the expansion culture in the step S3 in the preparation methods of the examples 2 to 6 is equivalent to that of the strain of the example 1; the gastric acid resistance of the biological agents prepared in examples 2 to 6 was comparable to that of the biological agent of example 1.
4. Choline tolerance assay
Taking bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri obtained by the expansion culture in the step S3 in the preparation method of the example 1 for choline tolerance measurement; and the biological agent finished product prepared in the example 1 is synchronously subjected to choline resistance measurement, and the results are shown in the table 4:
TABLE 4 Choline resistance measurement results
Testing 3000 strains of single bacteria; mixed bacteria 12000 strains were tested, containing 3000 strains each.
From the above results, it is clear that bifidobacterium animalis and lactobacillus plantarum have good survival rates in simulated bile; lactobacillus acidophilus and Lactobacillus reuteri have relatively low survival rates in simulated bile. The survival rate of the modified mixed bacteria in simulated bile is improved, the comprehensive survival rate can reach more than 70%, and the survival rate is greatly improved.
The choline resistance of the strain obtained by the expansion culture in the step S3 in the preparation methods of the examples 2 to 6 is equivalent to that of the strain of the example 1; the choline resistance of the biological agents prepared in examples 2 to 6 was comparable to that of the biological agent of example 1.
5. Experiment of anti-ulcerative colitis Effect
After 50 mice were selected for 2 months, and established a colitis model using DSS-induced mice, 0.2g of the biological agent prepared in example 1 was taken daily for intervention, 6 g/day was taken normally, the body weight of the mice was measured every two days, and the body weight change results after daily excretion are shown in table 5:
TABLE 5 results of weight change in mice
From the results, the biological preparation can effectively inhibit the weight loss of mice caused by DSS, and has good effects of improving intestinal flora and resisting ulcerative colitis.
Claims (5)
1. A preparation method of an acid and alkali resistant biological agent for resisting ulcerative colitis, which is characterized by comprising the following steps:
S1, preparing soybean protein: washing and soaking organic low-fat soybeans by adopting MRM molecular resonance, and then pulping and curing, defoaming, sterilizing and cooling to obtain soybean proteins;
S2, preparing a culture medium: adding 90 g-110 g g soybean protein, 90 g-110 g g glucose, 1.8 g-2.2 g sodium bicarbonate, 1.8 g-2.2 g vitamin C, 2.8 g-3.2 g sodium glutamate, 0.25 g-0.35 g sodium chloride and 1.2 g-1.8 g citric acid into each liter of water, and mixing to prepare a sterile culture medium;
S3, culturing: inoculating animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to a sterile culture medium respectively, and performing micro-dynamic fermentation culture under an MRM molecular resonance environment with the generation amount of negative ions of 900 e -/cm3 -1500 e -/cm3 respectively, so as to extract and obtain amplified cultured animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri;
S4, bacterial modification: spraying protective agents on the surfaces of bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri respectively, drying and then spraying film forming agents to obtain modified bacteria;
Wherein the protectant comprises purified water, soy protein, and beta-cyclodextrin; the film forming agent comprises enteric polyacrylic resin, hydroxypropyl methylcellulose phthalate, ethyl cellulose, polyethylene glycol, glycerol and triacetin;
S5, bacteria compounding: compounding the modified animal bifidobacterium, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri to obtain a biological preparation;
In S4, the components of the protective agent comprise, by mass, 80-100 parts of purified water, 15-25 parts of soybean protein and 0.5-1 part of beta-cyclodextrin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the protective agent=100 (3-5);
S4, the film forming agent comprises the following components in parts by mass: 15 to 30 parts of enteric polyacrylic resin, 15 to 30 parts of hydroxypropyl methylcellulose phthalate, 15 to 30 parts of ethylcellulose, 2 to 5 parts of polyethylene glycol, 2 to 5 parts of glycerol and 2 to 5 parts of triacetin; the spraying quantity of the spraying is that the mass ratio of the bacterial powder to the film forming agent=100 (3-5);
in S5, the mass ratio of the compound is that bifidobacterium animalis, lactobacillus plantarum, lactobacillus acidophilus and lactobacillus reuteri are (35-55) = (20-40): (5-15): (10-15);
in S5, nattokinase is also added into the biological agent, and the adding mass ratio is as follows: biological agent nattokinase=100 (6-10).
2. The method for preparing an acid and alkali resistant biological agent for treating ulcerative colitis according to claim 1, wherein in S1, the soaking time is 6 h-10 h; the sterilization adopts a pasteurization method; the cooling temperature is 38-42 ℃.
3. The preparation method of the acid and alkali resistant biological preparation for resisting ulcerative colitis of claim 1, wherein in S2, 25-35% of brown sugar by mass percent is contained in glucose, which is beneficial to the ingestion of amino acid by strains.
4. The method for preparing an acid and alkali resistant biological agent against ulcerative colitis according to claim 1, wherein in S3, the sterile medium is homogenized after inoculation, and the speed of homogenization is 45rpm to 55rpm.
5. The method for preparing an acid and alkali resistant biological agent for treating ulcerative colitis according to claim 1, wherein in S3, the temperature of the micro-dynamic fermentation culture is 39-42 ℃ and the time is 5 h-7 h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311755723.8A CN117801989B (en) | 2023-12-19 | 2023-12-19 | Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311755723.8A CN117801989B (en) | 2023-12-19 | 2023-12-19 | Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117801989A CN117801989A (en) | 2024-04-02 |
| CN117801989B true CN117801989B (en) | 2024-05-14 |
Family
ID=90421347
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311755723.8A Active CN117801989B (en) | 2023-12-19 | 2023-12-19 | Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117801989B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN118271921A (en) * | 2024-06-03 | 2024-07-02 | 沈阳盛达惠发化工有限公司 | Negative ion furniture paint and preparation and use methods thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988123A (en) * | 2018-01-17 | 2018-05-04 | 江南大学 | One plant has the lactobacillus plantarum for adjusting ampicillin induction enteric flora disturbance |
| CN110564638A (en) * | 2019-07-29 | 2019-12-13 | 江南大学 | Lactobacillus reuteri with probiotic characteristics and application thereof |
| CN113943687A (en) * | 2021-12-21 | 2022-01-18 | 山东中科嘉亿生物工程有限公司 | Lactobacillus reuteri JYLB-291 for improving ulcerative colitis and application thereof |
| WO2022144483A1 (en) * | 2020-12-30 | 2022-07-07 | Universitat De València (60%) | Lactobacillus plantarum strain, use as a probiotic and bioactive product derived therefrom |
| CN116790409A (en) * | 2023-03-22 | 2023-09-22 | 大连三仪动物药品有限公司 | Lactobacillus reuteri and preparation and application of microecological preparation thereof |
| CN116836852A (en) * | 2023-06-25 | 2023-10-03 | 浙江大学 | Lactobacillus reuteri ZJUIDS09 for relieving ulcerative colitis and application thereof |
-
2023
- 2023-12-19 CN CN202311755723.8A patent/CN117801989B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107988123A (en) * | 2018-01-17 | 2018-05-04 | 江南大学 | One plant has the lactobacillus plantarum for adjusting ampicillin induction enteric flora disturbance |
| CN110564638A (en) * | 2019-07-29 | 2019-12-13 | 江南大学 | Lactobacillus reuteri with probiotic characteristics and application thereof |
| WO2022144483A1 (en) * | 2020-12-30 | 2022-07-07 | Universitat De València (60%) | Lactobacillus plantarum strain, use as a probiotic and bioactive product derived therefrom |
| CN113943687A (en) * | 2021-12-21 | 2022-01-18 | 山东中科嘉亿生物工程有限公司 | Lactobacillus reuteri JYLB-291 for improving ulcerative colitis and application thereof |
| CN116790409A (en) * | 2023-03-22 | 2023-09-22 | 大连三仪动物药品有限公司 | Lactobacillus reuteri and preparation and application of microecological preparation thereof |
| CN116836852A (en) * | 2023-06-25 | 2023-10-03 | 浙江大学 | Lactobacillus reuteri ZJUIDS09 for relieving ulcerative colitis and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117801989A (en) | 2024-04-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1189560C (en) | Use of bacteria endowed with arginine deiminase to induce apoptosis and/or reduce an inflammatory reaction and pharmaceutical or dietetic compositions containing such bacteria | |
| USRE40023E1 (en) | Dietary and pharmaceutical compositions containing lyophilized lactic bacteria, their preparation and use | |
| KR100978648B1 (en) | A new bacillus subtilis strain and its use in preparing medicine for treating thrombosis | |
| DE10101793A1 (en) | Use of SLPI to treat inflammatory bowel disease | |
| CN117801989B (en) | Preparation method of acid and alkali resistant biological preparation for resisting ulcerative colitis | |
| CN112746034B (en) | Lactobacillus rhamnosus LR863 and lactobacillus rhamnosus LR519 for synergistically inhibiting helicobacter pylori and application thereof | |
| US20220378856A1 (en) | New probiotic composition for prevention of bacterial vaginosis | |
| KR102048690B1 (en) | lactic acid bacteria improved stability and preparing method thereof | |
| CN106860451A (en) | A kind of new opplication of rifamycin nitroimidazole coupling molecule | |
| CN117721033B (en) | Lactobacillus mucilaginosus KS6 and application thereof in preparation of anti-inflammatory and sleep-aiding foods and medicines | |
| US20180344783A1 (en) | Natural intra-vaginal inserts to control imbalanced ph | |
| Xu et al. | Enterococci facilitate polymicrobial infections | |
| KR101021226B1 (en) | Therapeutic usage of copa3 peptide analogue isolated from copris tripartitus for c. difficile-pseudomembranous colitis and general gut inflammation | |
| CN111567807A (en) | Rhamnose-containing lactobacillus composition for inhibiting inflammatory reaction and resisting vaginitis and application thereof | |
| CN113262242B (en) | Application of bifidobacterium lactis JYBR-190 in removing in-vivo heavy metal products | |
| CN114747770A (en) | Application of lactobacillus acidophilus in preparing food or medicine for preventing or treating antibiotic associated diarrhea | |
| KR20180070484A (en) | lactic acid bacteria improved stability and preparing method thereof | |
| Legaria et al. | Intra-peritoneal abscess after an abdominal hysterectomy involving Cutibacterium avidum (former Propionibacterium avidum) highly resistant to clindamycin | |
| CN108309969B (en) | Medical application of tectorigenin in treating necrotic enteritis of chicken | |
| CN114149483A (en) | Antibacterial peptide composition and application thereof | |
| Ma et al. | First isolation and identification of Shewanella xiamenensis from diseased Nile tilapia (Oreochromis niloticus L.) | |
| CN107115368B (en) | Pomegranate peel microecological preparation for preventing animal diarrhea and application thereof | |
| Gibson et al. | Recovery of a probiotic organism from human faeces after oral dosing | |
| CN116606761B (en) | Bifidobacterium animalis subspecies BLa19 capable of relieving rheumatoid arthritis and application thereof | |
| CN118460440B (en) | Composite probiotics for preventing and treating colonitis 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 |