CN111500505A - Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics - Google Patents
Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics Download PDFInfo
- Publication number
- CN111500505A CN111500505A CN202010432513.5A CN202010432513A CN111500505A CN 111500505 A CN111500505 A CN 111500505A CN 202010432513 A CN202010432513 A CN 202010432513A CN 111500505 A CN111500505 A CN 111500505A
- Authority
- CN
- China
- Prior art keywords
- heavy metal
- probiotics
- probiotic
- vivo
- isolated
- 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
- 239000006041 probiotic Substances 0.000 title claims abstract description 79
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 79
- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 59
- 238000000034 method Methods 0.000 title claims abstract description 30
- 210000001035 gastrointestinal tract Anatomy 0.000 title claims abstract description 26
- 238000001727 in vivo Methods 0.000 title claims abstract description 18
- 230000000529 probiotic effect Effects 0.000 claims abstract description 34
- 235000013618 yogurt Nutrition 0.000 claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 9
- 244000005700 microbiome Species 0.000 claims abstract description 9
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 6
- 238000012258 culturing Methods 0.000 claims abstract description 6
- 238000001179 sorption measurement Methods 0.000 claims abstract description 6
- 239000003963 antioxidant agent Substances 0.000 claims abstract description 5
- 238000009304 pastoral farming Methods 0.000 claims abstract description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 25
- 241000894006 Bacteria Species 0.000 claims description 18
- 238000004458 analytical method Methods 0.000 claims description 16
- 210000004185 liver Anatomy 0.000 claims description 14
- 230000008439 repair process Effects 0.000 claims description 14
- 241000191998 Pediococcus acidilactici Species 0.000 claims description 11
- 230000009467 reduction Effects 0.000 claims description 10
- 241001465754 Metazoa Species 0.000 claims description 6
- 229910052804 chromium Inorganic materials 0.000 claims description 6
- 244000005709 gut microbiome Species 0.000 claims description 6
- 229910052745 lead Inorganic materials 0.000 claims description 5
- 229910052753 mercury Inorganic materials 0.000 claims description 5
- 229910052759 nickel Inorganic materials 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 4
- 230000036542 oxidative stress Effects 0.000 claims description 4
- 208000027418 Wounds and injury Diseases 0.000 claims description 3
- 230000009286 beneficial effect Effects 0.000 claims description 3
- 208000014674 injury Diseases 0.000 claims description 3
- 230000004792 oxidative damage Effects 0.000 claims description 2
- 238000004321 preservation Methods 0.000 claims description 2
- 230000002000 scavenging effect Effects 0.000 claims description 2
- 230000003647 oxidation Effects 0.000 abstract description 14
- 238000007254 oxidation reaction Methods 0.000 abstract description 14
- 230000000968 intestinal effect Effects 0.000 abstract description 8
- 238000002360 preparation method Methods 0.000 abstract description 5
- 230000000813 microbial effect Effects 0.000 abstract description 3
- 239000011651 chromium Substances 0.000 description 32
- 241000282414 Homo sapiens Species 0.000 description 12
- 241000186660 Lactobacillus Species 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 229940039696 lactobacillus Drugs 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 230000002292 Radical scavenging effect Effects 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 238000012163 sequencing technique Methods 0.000 description 7
- 210000003608 fece Anatomy 0.000 description 6
- 239000000203 mixture Substances 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 238000011282 treatment Methods 0.000 description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 5
- 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 5
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 5
- 238000009592 kidney function test Methods 0.000 description 5
- 238000007449 liver function test Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 230000009102 absorption Effects 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000009534 blood test Methods 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 210000003734 kidney Anatomy 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 235000018102 proteins Nutrition 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 235000020183 skimmed milk Nutrition 0.000 description 4
- 210000000813 small intestine Anatomy 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 244000199885 Lactobacillus bulgaricus Species 0.000 description 3
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 description 3
- 206010067125 Liver injury Diseases 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000003064 anti-oxidating effect Effects 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000002738 chelating agent Substances 0.000 description 3
- 235000021316 daily nutritional intake Nutrition 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000001493 electron microscopy Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000002550 fecal effect Effects 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 210000000936 intestine Anatomy 0.000 description 3
- 238000002955 isolation Methods 0.000 description 3
- 229940004208 lactobacillus bulgaricus Drugs 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 238000005067 remediation Methods 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- 235000020255 yak milk Nutrition 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- VYZAMTAEIAYCRO-UHFFFAOYSA-N Chromium Chemical compound [Cr] VYZAMTAEIAYCRO-UHFFFAOYSA-N 0.000 description 2
- 241000192125 Firmicutes Species 0.000 description 2
- 240000006024 Lactobacillus plantarum Species 0.000 description 2
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 2
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 235000015063 acidophilus milk Nutrition 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000010411 cooking Methods 0.000 description 2
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- -1 flavoreductase Proteins 0.000 description 2
- 231100000234 hepatic damage Toxicity 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 229940072205 lactobacillus plantarum Drugs 0.000 description 2
- 230000008818 liver damage Effects 0.000 description 2
- 230000003908 liver function Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 231100000783 metal toxicity Toxicity 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 229910017604 nitric acid Inorganic materials 0.000 description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000010183 spectrum analysis Methods 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 241000589220 Acetobacter Species 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 1
- 108010082126 Alanine transaminase Proteins 0.000 description 1
- 208000006820 Arthralgia Diseases 0.000 description 1
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 1
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- 241000605059 Bacteroidetes Species 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 229920000742 Cotton Polymers 0.000 description 1
- 101150027068 DEGS1 gene Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241001522878 Escherichia coli B Species 0.000 description 1
- 108010074122 Ferredoxins Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 240000008397 Ganoderma lucidum Species 0.000 description 1
- 235000001637 Ganoderma lucidum Nutrition 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 240000001046 Lactobacillus acidophilus Species 0.000 description 1
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 241000186840 Lactobacillus fermentum Species 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 208000005374 Poisoning Diseases 0.000 description 1
- 241000692844 Prevotellaceae Species 0.000 description 1
- 208000001647 Renal Insufficiency Diseases 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 206010053615 Thermal burn Diseases 0.000 description 1
- 101710172405 Thiol peroxidase Proteins 0.000 description 1
- 102000002933 Thioredoxin Human genes 0.000 description 1
- 238000008050 Total Bilirubin Reagent Methods 0.000 description 1
- 241000609666 Tuber aestivum Species 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 239000005862 Whey Substances 0.000 description 1
- 102000007544 Whey Proteins Human genes 0.000 description 1
- 108010046377 Whey Proteins Proteins 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 208000005882 cadmium poisoning Diseases 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 108010071930 chromate reductase Proteins 0.000 description 1
- 230000010428 chromatin condensation Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 229940109239 creatinine Drugs 0.000 description 1
- 230000001086 cytosolic effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 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 1
- 210000003372 endocrine gland Anatomy 0.000 description 1
- IINNWAYUJNWZRM-UHFFFAOYSA-L erythrosin B Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 IINNWAYUJNWZRM-UHFFFAOYSA-L 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 239000010842 industrial wastewater Substances 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 201000006370 kidney failure Diseases 0.000 description 1
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 229940012969 lactobacillus fermentum Drugs 0.000 description 1
- 230000002045 lasting effect Effects 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 230000030363 nerve development Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 231100000572 poisoning Toxicity 0.000 description 1
- 230000000607 poisoning effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000005057 refrigeration Methods 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009666 routine test Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000012064 sodium phosphate buffer Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 235000021262 sour milk Nutrition 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000012916 structural analysis Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 108060008226 thioredoxin Proteins 0.000 description 1
- 229940094937 thioredoxin Drugs 0.000 description 1
- 238000004627 transmission electron microscopy Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Images
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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
-
- 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
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/02—Antidotes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
- C12N1/02—Separating microorganisms from their culture media
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/41—Pediococcus
- A23V2400/413—Acidilactici
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Microbiology (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides an intestinal tract repairing method for reducing heavy metal residue in vivo by using probiotics, which is characterized by comprising the following steps: (1) separating microorganisms from yogurt in the grazing land of Qinghai-Tibet plateau; (2) preparing an MRS culture medium and a PBS solution; (3) inoculating and culturing; (4) detecting strains; (5) analyzing the antioxidant capacity of the probiotics; (6) analyzing the heavy metal resistance of the probiotics; (7) and (4) analyzing the heavy metal adsorption capacity of the probiotics. The method applies the probiotics with strong oxidation resistance to the preparation of the yoghourt, and reduces the oxidation pressure in the intestinal tract by eating the probiotic yoghourt, thereby protecting the intestinal flora and leading the microbial community to better play a role in reducing the heavy metal residue.
Description
Technical Field
The invention relates to the field of microbial genetic engineering, in particular to a probiotic strain and an intestinal tract repair method for reducing heavy metal residues in vivo by using the probiotic strain.
Background
Heavy metal contamination has become a global problem. In recent years, under the influence of discharge of heavy metal industrial wastewater and daily activities of human beings, about 46700hm is generated every year in China2The land is contaminated with heavy metals. Heavy metals enter human bodies through food chains, and pose serious threats to human health, for example, cadmium poisoning can cause diseases such as osteoporosis, arthralgia and renal failure, and chromium poisoning can cause damage to liver, kidney and endocrine glands, so that the diseases are diseased. At present, heavy metal pollution in soil is mainly carried out by methods such as physical remediation, chemical remediation, phytoremediation and the like, but the methods have the problems of time consumption, high cost, secondary pollution and the like. In addition, the existing heavy metal restoration is mainly carried out in the environment, and the restoration area of the methods is extremely limited in consideration of the large-area heavy metal pollution range in the world, so that the human beings are still inevitably threatened by the heavy metals. Therefore, a safe and effective heavy metal remediation method is urgently needed to be found.
The intestinal tract is the largest immune organ of the human body, and has important functions of improving the immunity of the human body besides functions of digestion, absorption and secretion. At least 1000 different types of microorganisms exist in the intestinal tract of a human body, and the microorganisms play an important role in regulating the functions of the intestinal tract, and researches show that sterile mice have nerve development defects, poor lymph development and low immune function compared with normal mice. Intestinal microorganisms affect various physiological activities such as development, absorption, immunity and the like of the intestinal tract. Thus, the function of the gut can be improved by modulating the gut microflora composition.
The probiotics are active microorganisms which colonize in human intestinal tracts and are beneficial to organisms, and the Tibetan plateau probiotics can not only directly repair heavy metal toxicity, but also indirectly repair the heavy metal toxicity by adjusting (/ remodeling) intestinal microflora. The method for repairing the intestinal tract by using the probiotics is safe and effective, simple and convenient to operate, low in price and long in lasting action time, not only can reduce the residue of heavy metal in a body, but also can enhance the immunity of a human body by regulating the composition and metabolism of intestinal flora.
The problems existing in the prior art are as follows:
(1) at present, in clinical practice, the method for detoxifying human body weight metal mainly uses a chelating agent for treatment, but the method has large side effects, such as the risks of carcinogenesis and teratogenesis, and simultaneously, due to the difficult degradability of most chelating agents, secondary pollution is caused to the environment after the chelating agents are discharged from the body through intestinal tracts.
(2) At present, extracts with oxidation resistance are also utilized to play a role in oxidation resistance, for example, extracts of fungi such as ganoderma lucidum spore powder and truffle have strong oxidation resistance; the plant extracts of ramulus Mori, folium Pruni and folium Ziziphi Spinosae also have antioxidant effect, and can be administered to reduce oxidation pressure in intestinal tract. However, compared with probiotics, the extraction process of the extract is more complicated, the preparation cost is higher, the acting time in the gastrointestinal tract is shorter, and the extract needs to be eaten for a long time.
(3) In the existing method for reducing the residual of heavy metals in vivo by using probiotics, the selected strain basis is the adsorption effect on the heavy metals, and a single strain is aimed at a single heavy metal. But the environment is polluted by a plurality of heavy metals at present, so the action range of removing the heavy metals by the existing probiotics is smaller.
Disclosure of Invention
In view of the defects of the prior art, the invention provides an intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics. A new strain is screened out, and the intestinal tract repairing method for reducing heavy metal residues in vivo by using the strain is provided. The invention adopts the following technical scheme:
1. a probiotic strain capable of reducing heavy metal residues in vivo, wherein the probiotic strain is Pediococcus acidilactici (Pediococcus acidilactici) QZ-01, which is deposited in China Center for Type Culture Collection (CCTCC) at 11 months and 14 days in 2019, and the address is as follows: in Wuhan university school of Wuhan 299 in Wuchang district of Wuhan city, Hubei province, the preservation number is CCTCCNO: m2019930.
2. A separation method of probiotics capable of reducing heavy metal residues in vivo comprises the following steps:
(1) separating microorganisms from yogurt in the grazing land of Qinghai-Tibet plateau; (2) preparing an MRS culture medium and a PBS solution; (3) inoculating and culturing; (4) detecting strains; (5) analyzing the antioxidant capacity of the probiotics; (6) analyzing the heavy metal resistance of the probiotics; (7) and (4) analyzing the heavy metal adsorption capacity of the probiotics.
3. And (3) comprehensively selecting a strain pediococcus acidilactici QZ-01 with high heavy metal resistance and oxidation resistance from the screened probiotics for preparing the functional yogurt in the next step.
4. A method for preparing yoghourt for reducing heavy metal residues in vivo comprises the following steps:
(1) activation of probiotic strains; (2) fermenting and making yoghourt; (3) and (4) carrying out experimental analysis on the yoghourt animal.
5. The application of the probiotics QZ-01 in reducing heavy metals in vivo comprises but is not limited to Cr, Hg, Pb and Ni.
6. Use of probiotic QZ-01 for scavenging hydroxyl and DPPH radicals.
7. Application of probiotic QZ-01 in adsorption of Cr, Hg, Pb and Ni.
8. Application of probiotic QZ-01 in repairing of Cr-induced liver oxidative stress and injury.
9. The application of the probiotics QZ-01 in reducing the abundance of harmful bacteria and increasing the abundance of beneficial bacteria.
10. The probiotic QZ-01 can be used for up-regulating and expressing the genes related to Cr reduction and oxidation resistance in the intestinal microflora.
11. The probiotic QZ-01 up-regulates the Cr resistance and the reduction capability of the expressed gene with unknown function.
12. Application of probiotic QZ-01 in reducing intestinal tract repair of heavy metal residues in vivo.
Has the advantages that:
the application provides a new bacterial strain, and is applied to the preparation of acidophilus milk through this bacterial strain, through edible probiotic acidophilus milk reduction oxidation pressure in the intestinal to protection intestinal flora makes better performance of microbial community reduce the effect that the heavy metal is remained, and this kind of method has the broad spectrum nature to the getting rid of heavy metal, and it is effectual to get rid of, easy operation, and the low price, and can establish advantages such as continuously playing the effect in the intestinal.
Drawings
FIG. 1 shows the results of the isolation of probiotics according to the invention;
FIG. 2 shows the preparation result of the yogurt of the present invention;
FIG. 3 shows the hydroxyl radical and DPPH radical scavenging ability of different probiotics according to the invention;
among them, a hydroxyl radical (A) and a DPPH radical (B).
FIG. 4 shows the results of the resistance of different probiotics of the present invention to heavy metals Cr, Hg, Pb, Ni;
wherein, Cr (A), Hg (B), Pb (C), Ni (D).
FIG. 5 shows experimental groupings of animals according to the present invention;
FIG. 6A shows the daily food intake of a mouse according to the present invention; b is the influence of Cr (VI) and probiotics on the growth rate of the mouse;
FIG. 7 is a graph of the effect of probiotic QZ-01 and XS40 treatment of the invention on the chromium content in mouse feces and tissues;
among these, mouse feces (a) and tissues (B).
FIG. 8 shows the repairing effect of the probiotics QZ-01 and XS40 of the invention on the liver injury of mice caused by Cr (VI);
FIG. 9A is a graph showing the results of a quantitative study of QZ-01 in feces from probiotic QZ-01 treated groups of the present invention; b is the content of QZ-01 in the excrement after 4 different treatments for 20 days; c and D are the reducing capacity of fecal culturable microorganisms to Cr (VI) at 37 ℃ for 72 hours and 44 hours, respectively;
FIG. 10 is the composition of total bacteria and metabolically active bacteria in feces of the invention;
wherein (A) the ratio of firmicutes to bacteroidetes; (B) the relative abundance of the bacterial 16SrRNA gene at the family level; (C) genus level heatmap based on 16S rRNA sequencing; (D) the classification assignment of mRNA at the genus level for each sample.
FIG. 11 is an analysis of the resistance and reducing power of the probiotic QZ-01 of the present invention to Cr (VI) of an unknown functional gene with increased differential expression of gut microbes;
the method comprises the following steps of (A) distribution of Cr repair related genes in the DEGS, (B) growth of engineering escherichia coli strains in L B culture medium containing 1Mm Cr (VI), (C) Cr (VI) reduction rate of engineering escherichia coli B L21 containing GCrR genes, (D) transmission electron microscopy and energy spectrum analysis of engineering escherichia coli treated and untreated with 1mM Cr (VI), (E) evolutionary relationship of GCrR and known chromate reductase of different genera, (F) GCrR homologous sequence analysis, and (G) structural modeling of 53431.
FIG. 12 is a schematic illustration of the mechanism of "bowel repair" according to the present invention;
FIG. 13 is a report of liver and kidney function test of the same school A;
FIG. 14 is a report of routine blood tests of the same school of A.A. in accordance with the present invention;
FIG. 15 is a report of the liver function and kidney function test of the same student class B; (ii) a
FIG. 16 is a report of routine blood tests of the same school according to the present invention;
FIG. 17 is the liver and kidney function test report of the third school student of the invention
FIG. 18 is a report of a routine blood test of the third school of the invention;
FIG. 19 is a report of liver and kidney function tests of Ding classmate in accordance with the present invention;
FIG. 20 is a report of a routine test of blood of the same school according to the present invention;
FIG. 21 is a report of liver and kidney function tests of the same student of the invention;
FIG. 22 is a report of routine blood tests of the pentastudy of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, embodiments of the present invention are described in detail below with reference to the accompanying drawings. Examples of these preferred embodiments are illustrated in the accompanying drawings. The embodiments of the invention shown in the drawings and described in accordance with the drawings are exemplary only, and the invention is not limited to these embodiments.
It should be noted that, in order to avoid obscuring the technical solution of the present invention with unnecessary details, only the structures and/or processing steps closely related to the solution according to the present invention are shown in the drawings, and other details that are not relevant are omitted.
Example 1
The embodiment provides a separation method capable of reducing in-vivo heavy metal residual probiotics, which comprises the following steps:
1. isolation of microorganisms from yogurt from the grazing land of Qinghai-Tibet plateau
The yogurt is prepared by herdsmen on Qinghai-Tibet plateau. Firstly, collecting fresh yak milk, pouring the yak milk into a pot, cooking with small fire, wherein the temperature cannot be too high and the yak milk cannot scald the hands, and cooking for later use. Taking a proper amount of the prepared yoghourt in the previous day, adding the prepared yoghourt into the boiled fresh milk, uniformly stirring, and then fermenting for 3-4 hours at the temperature of about 42 ℃.
2. MRS culture medium and PBS solution are prepared
Putting 63.3g MRS agar into conical flask, adding 1000m L distilled water, sealing with cotton plug, sterilizing at 121 deg.C for 20min, respectively weighing 0.8g NaCl, 0.02g KCl, and 0.364g Na2HPO4·12H2O、0.024g KH2PO4Placing into a conical flask, adding 100m L distilled water, stirring, adjusting pH to 7.4, and sterilizing.
3. Inoculation and culture
Pouring the sterilized MRS solid culture medium into culture dishes in a clean bench, pouring about 30m L into each culture dish, sealing a sealing film, inverting for later use, sucking 5m L PBS solution by using a pipette, adding the PBS solution into a test tube, mixing a small amount of yoghourt into the PBS solution for dilution, coating the diluted yoghourt on the MRS solid culture medium, and culturing at the constant temperature of 37 ℃ for 18 hours.
4. Strain detection
As shown in the attached FIG. 1 and Table 1, single colonies were obtained, and then single colonies were picked for colony PCR using 27F/1492R universal primers, dNTP mix, Taq enzyme, Taq Buffer and other reagents required for PCR were purchased from Takara Bio Inc., and the amplification procedures were 94 ℃ for 5min, (95 ℃ for 30S, 58 ℃ for 30S, 72 ℃ for 90S) for 35 cycles, and 72 ℃ for 10 min. the amplified samples were subjected to 16S rRNA sequencing, and the strains selected are shown in Table 2, including Pediococcus acidilactici (Pediococcus acidilactici), Lactobacillus fermentum (L actans fermentum), Lactobacillus plantarum (L Acobacterium), Lactobacillus acidophilus (L Acidophilus), Lactobacillus casei (L Acobacterium ei), Lactobacillus bulgaricus (L Lactobacillus bulgaricus) and the like.
5. Antioxidant capacity analysis of probiotics
The hydroxyl radical scavenging capacity of several probiotics is determined (1M L0.75 mM phenanthroline, 1.5M L0.15.15M sodium phosphate buffer and 1M L0.75 mM FeSO are respectively taken4And 1m L0.01% H2O2Preparing mixed solution, adding sample to be tested 1m L, mixing, incubating at 37 deg.C for 30min, and measuring absorbance at 536nm2O2Absorbance of (b). Hydroxy radical scavenging ability ═ aSample (I)-ABlank space]/[AControl-ABlank space]) DPPH radical scavenging ability (1 m L sample and 1m L0.2.2 mM DPPH solution (dissolved in methanol) were mixed and reacted in the dark for 30 minutes, using phosphate buffer as a blank, and the absorbance value of the mixture at 517nm was measuredBlank space-ASample (I)]/ABlank space) As shown in fig. 3: the hydroxyl radical scavenging ability and DPPH radical scavenging ability of QZ-01 are the highest, wherein the hydroxyl radical scavenging rate can reach more than 80%, and DPPH radical scavenging rate can reach more than 50%.
6. Analysis of heavy metal resistance of probiotics
Probiotics screened by MRS activationBacteria; ni to be sterilized2+、Cd2+、Cr6+、Pb2+Adding into culture medium respectively; inoculating 1% of the activated strain into each medium; the cells were incubated overnight at a constant temperature of 37 ℃ on a shaker at 150r/min and then measured for OD600 using visible light spectrophotometry, with 3 replicates per set of experiments. As shown in FIG. 4, QZ-01 is most resistant to several heavy metal ions.
7. Analysis of heavy metal adsorption Capacity of Probiotics
Culturing probiotic bacteria with MRS culture medium at 37 deg.C for 18h, centrifuging respectively, measuring dry weight and wet weight of bacteria, suspending probiotic bacteria cells with PBS buffer solution to make bacteria concentration 5 g/L (bacteria wet weight), and adding Ni into the bacteria suspension respectively2+、Cd2+、Cr6 +、 Pb2+Adjusting pH to 6.28 deg.C with NaOH and HC L to reach a final concentration of 20 mM., culturing for 2h, centrifuging, discarding supernatant, collecting precipitate, digesting with mixed solution of concentrated nitric acid and perchloric acid (volume ratio of 4: 1) at 300 deg.C for 10min, and determining the content of each heavy metal by flame atomic absorption.
The results show that: and comprehensively selecting a strain pediococcus acidilactici QZ-01 with high heavy metal resistance, high oxidation resistance and high oxidation resistance from the screened probiotics for the next step of making the functional yogurt.
Example 2
The embodiment provides a method for preparing yoghourt by reducing heavy metal residues in vivo, which comprises the following steps:
1. activation of probiotic bacterial strains
Inoculating the probiotic strain preserved on the MRS slant to skim milk (purchased from American BD company) (milk powder: water 1:8, w/v) to culture at 37-42 ℃, and performing first activation, wherein the culture time is determined according to the specific curd condition, and the culture is completed when the yoghourt is in a uniform solidification state and a small amount of whey is separated out on the surface of the curd.
2. Yoghourt fermentation preparation
Adding 3-10% of sucrose into the prepared skim milk, preheating for 10min at 50 ℃, then sterilizing at 95 ℃ for 5min, and cooling the skim milk to 42 ℃. Inoculating activated Pediococcus acidilactici, Lactobacillus plantarum, Streptococcus thermophilus and Lactobacillus bulgaricus to skim milk according to 2% -5%, and fermenting at 42 deg.C for 4-6 h. After-ripening for 12h at 4 ℃ by refrigeration. The prepared yogurt is shown in figure 2.
3. Experimental analysis of sour milk animals
Animal experiment groups are shown in figure 5, prepared yogurt is intragastrically administered to mice with an intragastric syringe needle, and the intragastric administration is continued for 30 days with 1g of yogurt per day (1 g of yogurt is dissolved in 500ul of sterile water).
(1) Daily food intake and growth rate analysis of mice
Mouse body weight and food intake were recorded weekly. The results are shown in figure 6, and the probiotics have no obvious influence on the daily food intake of the mice, but improve the growth rate of the mice.
(2) Ability of probiotics to remove heavy metals
Dissecting mouse, collecting organs (including liver, kidney, and small intestine), digesting the organs with concentrated nitric acid, and measuring heavy metal content in the organs by flame atomic absorption method. As shown in fig. 7, the heavy metal content in the liver of the mouse is reduced by about 46%, the heavy metal content in the kidney is reduced by about 43%, the heavy metal content in the small intestine is reduced by about 88%, and the heavy metal content in the feces is increased by about 27%, which indicates that the probiotic bacteria effectively reduce the heavy metal residue in the mouse.
(3) Repair effect of probiotics on mouse liver oxidation pressure and injury caused by Cr (VI)
Analysis of the repair ability of probiotics to liver and small intestine showed that pediococcus acidilactici reversed the change in the cr (vi) -induced oxidative stress marker in figure 8. Cr (vi) exposure can cause significant liver damage, including cytoplasmic vacuolization, nuclear pyknosis, and chromatin condensation, which can be mitigated by pediococcus acidilactici. Shows that QZ-01 has obvious inhibition effect on the inflammatory reaction induced by Cr (VI).
(4) Analysis of reducing power of probiotics on Cr (VI)
As shown in figure 9A, the QZ-01 begins to be planted on the 8 th day, and tends to be stable after the planting amount reaches the maximum on the 20 th day, and as shown in figure 9B, the QZ-01 in the feces of different treatment groups is quantitatively found, and the QZ-01 can be effectively planted in the intestinal tract. As shown in FIGS. 9C and 9D, analysis of the reducing power of Cr (VI) in the flora in the fecal samples of the different treatment groups showed that QZ-01 significantly improved the reducing power of Cr (VI) in the fecal flora.
(5) Analysis of gut microflora composition by 16S rRNA sequencing and Macro-transcriptome sequencing
As shown in fig. 10A, at the phylum level, the proportion of Firmicutes to Bacteroides was significantly different between the different sequencing methods, 16S rRNA sequencing analysis showed no significant difference in this proportion between the different treatment groups, macrotranscriptome sequencing analysis showed that this proportion was significantly higher in the QZ-01 repair group than in the other groups, which showed that active metabolic bacteria or play an important role in gut repair at the family level, 16S rRNA analysis showed S24-7 (12.47-42.03%), Prevotellaceae (10.88-36.07%), bacteroidacedaceae (4.57-19.66%), L acetobacter ceraceae (2.92-15.20%) and paramaprezotellaceae (0.46-15.37%) higher abundance (fig. 10B) heat map at the genus level showed that the colony of the qcr-01 repair group was close to the colony of the control group, the colony structure was significantly more relieved from the lactobacillus group, the lactobacillus group was significantly restored from the probiotic group, as shown by the relative change in fig. 0.01 to lactobacillus (0.01-7), the relative abundance was significantly increased from the corresponding to the corresponding intestinal tract infection of bacillus (0.8, the lactobacillus) and the corresponding recovery of the lactobacillus group (20, the corresponding to the corresponding intestinal tract) was significantly increased from the corresponding to the corresponding recovery of bacillus (20% of lactobacillus) (fig. 8, 7, 8-8, 8-8, 8-8, 8-8, and 8-8, 8-8% of lactobacillus-8 (7) and 8-8 (7) and similar to the corresponding recovery of lactobacillus group of lactobacillus-8 (7, and similar to the corresponding intestinal tract infections of all of lactobacillus group of lactobacillus).
(6) Probiotics QZ-01 up-regulate expression of genes related to Cr (VI) reduction and antioxidation in intestinal microflora
As shown in table 2, thioredoxin, desulfurated ferredoxin, flavoreductase, FAD/nad (p) -bound oxidoreductase, and the like, which are involved in cr (vi) reduction, are expressed in the probiotic QZ-01 up-regulated compared to the cr (vi) group. Major erythrosin protein family proteins related to oxidative stress, thiol peroxidase, erythroredoxin, peroxidase, superoxide dismutase, catalase, and the like.
(7) Analysis of Cr (VI) resistance and reducing power of QZ-01 Up-regulated expressed functionally unknown Gene
As shown in fig. 11A, 788 genes whose expression is up-regulated in QZ-01Cr (vi) were selected from 57248 single genes, including 20 known Cr (vi) reduction-related genes, 14 known anti-oxidation-related genes, 546 other annotated genes and 208 genes with unknown functions, fig. 11B is a graph in which 10 genes selected at random from 208 genes with higher abundance are subjected to gene synthesis and a corresponding e.coli engineered bacterium (uns 1-10) is constructed, and the results show that 5 of the engineered bacteria ( uns 2,6,7,8,9) significantly improve resistance to Cr (vi), it is further found that the engineered bacterium e.coli uns 9/GCrR has a stronger reducing ability to Cr (vi) (fig. 11C) is subjected to transmission scanning on e.coli GCrR, and the energy spectrum analysis shows that intracellular Cr (iii) precipitates can be analyzed in GCrR strain gcr 3, the results show that the results are expressed by electron microscopy, see that the map 11A structural analysis of gcr 3, 11B is performed by electron microscopy, 11B is a graph in which shows that the amino acid sequence comparison of gcr 3, 11B is carried out from gcr 14 Cr (gcr) and 11G — 7, 9) is carried out by electron microscopy, and 11B (gcr iii) comparison of the known amino acid reduction system, and 11B).
(8) Mechanism of intestinal tract repair with probiotics
As shown in fig. 12, cr (vi) entering the animal body increases the oxidation pressure of the intestine and liver, increases cr (vi) residue in the small intestine, liver and kidney, causes liver damage and causes intestinal flora disorder, while probiotics can effectively colonize the intestine and reduce the oxidation pressure of the intestine, protect the intestinal flora to better function, reduce cr (vi) and reduce heavy metal residue in the body, thereby protecting the liver.
(9) Safety verification of human body experiment
The normal ranges of aspartate aminotransferase and alanine aminotransferase are 1-49U/L, the normal range of total protein is 60-82U/L, the normal range of total bilirubin is 9.1-30.1U/L, as shown in figures 13, 15, 17, 19 and 21, the student test results are all in the normal range, therefore, the edible probiotic yoghourt does not damage the liver function of the human body, the normal range of urea is 2.9-7.1U/L, the normal range of creatinine is 44-108U/L, the student report results are all normal, the normal range of erythrocyte is 4-1010 ^ 9/L, the normal range of leukocyte is 3.5-5.010 ^ 12/L, the normal range of protein is 110 g/150 g/L, and the student test results of erythrocyte and the student test results are all in the normal range of 18, 18 g/3514, 18 g/18, 18 g, 14 g, 84 g, 14.
The foregoing is directed to embodiments of the present application and it is noted that those skilled in the art may make various changes and modifications without departing from the spirit and scope of the application.
TABLE 1 isolation information of the strains
TABLE 2 probiotic QZ-01 Up-regulated expression and reduction-related genes, antioxidation-related genes and partial function-unknown genes of Cr (VI)
Claims (10)
1. A probiotic strain capable of reducing heavy metal residues in vivo is Pediococcus acidilactici (Pediococcus acidilactici) QZ-01, and is preserved in CCTCC at 11 months and 14 days in 2019 with the preservation number of M2019930.
2. The method for separating probiotics capable of reducing heavy metal residues in vivo according to claim 1, comprising the following steps: (1) separating microorganisms from yogurt in the grazing land of Qinghai-Tibet plateau; (2) preparing an MRS culture medium and a PBS solution; (3) inoculating and culturing; (4) detecting strains; (5) analyzing the antioxidant capacity of the probiotics; (6) analyzing the heavy metal resistance of the probiotics; (7) and (4) analyzing the heavy metal adsorption capacity of the probiotics.
3. A method for preparing yoghourt for reducing heavy metal residues in vivo comprises the following steps: (1) activation of probiotic strains; (2) fermenting and making yoghourt; (3) and (4) carrying out experimental analysis on the yoghourt animal.
4. The use of probiotic QZ-01 isolated by the method of claim 1 for reducing heavy metals in vivo, comprising: cr, Hg, Pb, Ni.
5. Use of probiotic bacteria QZ-01 isolated by the method of claim 1 for scavenging hydroxyl radicals and DPPH radicals.
6. The use of the probiotic QZ-01 isolated by the method of claim 1 for adsorbing Cr, Hg, Pb and Ni.
7. Use of probiotic bacteria QZ-01 isolated according to the method of claim 1 for Cr-induced liver oxidative stress and injury repair.
8. The use of the probiotic QZ-01 isolated by the method of claim 1 to reduce the abundance of harmful bacteria while increasing the abundance of beneficial bacteria.
9. The use of the probiotic QZ-01 isolated according to the method of claim 1 to up-regulate genes associated with Cr reduction and antioxidant in the expressed intestinal microflora.
10. Use of probiotic QZ-01 isolated according to the method of claim 1 for reducing heavy metal residuals in the body in intestinal tract repair.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010432513.5A CN111500505A (en) | 2020-05-20 | 2020-05-20 | Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010432513.5A CN111500505A (en) | 2020-05-20 | 2020-05-20 | Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111500505A true CN111500505A (en) | 2020-08-07 |
Family
ID=71868390
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010432513.5A Pending CN111500505A (en) | 2020-05-20 | 2020-05-20 | Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111500505A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113528400A (en) * | 2021-08-18 | 2021-10-22 | 兰州大学 | Lactobacillus fermentum with hexavalent chromium ion reducing capability and application thereof |
| CN113528399A (en) * | 2021-08-18 | 2021-10-22 | 兰州大学 | Pediococcus acidilactici with hexavalent chromium ion reducing capability and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109234189A (en) * | 2018-08-17 | 2019-01-18 | 甘肃普诺贝康生物科技有限责任公司 | One plant of lactobacillus plantarum strain BX62 and its application with oxidation resistance |
| CN111109359A (en) * | 2019-12-29 | 2020-05-08 | 江南大学 | Multifunctional pediococcus acidilactici CCFM1105, fermented food thereof and application |
-
2020
- 2020-05-20 CN CN202010432513.5A patent/CN111500505A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109234189A (en) * | 2018-08-17 | 2019-01-18 | 甘肃普诺贝康生物科技有限责任公司 | One plant of lactobacillus plantarum strain BX62 and its application with oxidation resistance |
| CN111109359A (en) * | 2019-12-29 | 2020-05-08 | 江南大学 | Multifunctional pediococcus acidilactici CCFM1105, fermented food thereof and application |
Non-Patent Citations (2)
| Title |
|---|
| BAO LE等: "Biosorption of cadmium by potential probiotic Pediococcus pentosaceus using in vitro digestion model", 《BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY》 * |
| 李畅: "具有潜在益生特性的乳酸菌对铅吸收特性及吸附机制的研究", 《中国优秀硕士学位论文全文数据库（工程科技I辑）》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113528400A (en) * | 2021-08-18 | 2021-10-22 | 兰州大学 | Lactobacillus fermentum with hexavalent chromium ion reducing capability and application thereof |
| CN113528399A (en) * | 2021-08-18 | 2021-10-22 | 兰州大学 | Pediococcus acidilactici with hexavalent chromium ion reducing capability and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Dordević et al. | Hydrogen sulfide toxicity in the gut environment: Meta-analysis of sulfate-reducing and lactic acid bacteria in inflammatory processes | |
| Shehata et al. | Screening of isolated potential probiotic lactic acid bacteria for cholesterol lowering property and bile salt hydrolase activity | |
| Zhai et al. | Screening of lactic acid bacteria with potential protective effects against cadmium toxicity | |
| Banwo et al. | Binding capacities and antioxidant activities of Lactobacillus plantarum and Pichia kudriavzevii against cadmium and lead toxicities | |
| EP2833898B1 (en) | Food grade bacteria for the removal of toxic compounds | |
| CN111040960B (en) | Lactobacillus plantarum LP33 and application thereof in preparation of product for promoting lead excretion | |
| CN111096459B (en) | Application of lactobacillus plantarum LP33 in preparation of product for preventing lead poisoning | |
| CN111500505A (en) | Intestinal tract repairing method for reducing heavy metal residues in vivo by using probiotics | |
| Alizadeh et al. | Mitigation of potentially toxic elements in food products by probiotic bacteria: a comprehensive review | |
| Murtaza et al. | In-vitro assessment of a novel plant rhizobacterium, Citrobacter freundii, for degrading and biocontrol of food mycotoxin deoxynivalenol | |
| Bhakta et al. | Probiotic characterization of arsenic-resistant lactic acid bacteria for possible application as arsenic bioremediation tool in fish for safe fish food production | |
| Fazeli et al. | Reduction in serum lipid parameters by incorporation of a native strain of Lactobacillus Plantarum A7 in Mice | |
| Wang et al. | Bioremediation of selenium-contaminated soil using earthworm Eisenia fetida: Effects of gut bacteria in feces on the soil microbiome | |
| El-Meihy et al. | Characterization and identification of cadmium-tolerant bacteria isolated from contaminated regions in Egypt | |
| CN106399153A (en) | Bacillus with tolerance to vanadium and use thereof | |
| CN117551557A (en) | Low-temperature-resistant multifunctional bacterium and application thereof in soil and water pollution remediation | |
| CN116769655A (en) | Streptococcus salivarius thermophilus JIAN+ and application thereof | |
| AU2014254812B2 (en) | Bacterium belonging to genus Lactobacillus | |
| Ahire | Studies on probiotic microorganism (s) and its biogenic metabolite (s) | |
| Micsinai et al. | Rhizome-associated bacterial communities of healthy and declining reed stands in Lake Velencei, Hungary | |
| Alrabadi et al. | Lactobacillus rhamnosus ability of aflatoxin detoxification | |
| CN110055186B (en) | Microbial preparation for degrading vomitoxin and application | |
| El-Meihy | Isolation and characterization of zinc tolerant bacteria from contaminated sediments and soils in Egypt | |
| Selta et al. | Bacillus siamensis I7B strain as heavy metal quencher and probiotics; isolated from gut of Stolepherous commersonni | |
| Bayane et al. | Assessment of the physiological and biochemical characterization of a Lactic acid bacterium isolated from chicken faeces in sahelian region |
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 |