CN116355812A - Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products - Google Patents
Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products Download PDFInfo
- Publication number
- CN116355812A CN116355812A CN202310530608.4A CN202310530608A CN116355812A CN 116355812 A CN116355812 A CN 116355812A CN 202310530608 A CN202310530608 A CN 202310530608A CN 116355812 A CN116355812 A CN 116355812A
- Authority
- CN
- China
- Prior art keywords
- reuteri
- lactobacillus reuteri
- hclr01
- limosilactobacillus
- cell lysate
- 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
- 241000186604 Lactobacillus reuteri Species 0.000 title claims abstract description 94
- 229940001882 lactobacillus reuteri Drugs 0.000 title claims abstract description 94
- 230000003712 anti-aging effect Effects 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title abstract description 11
- 239000013592 cell lysate Substances 0.000 claims abstract description 34
- 239000007788 liquid Substances 0.000 claims description 22
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 238000000502 dialysis Methods 0.000 claims description 20
- 239000012528 membrane Substances 0.000 claims description 14
- 238000002156 mixing Methods 0.000 claims description 13
- 239000011148 porous material Substances 0.000 claims description 12
- 239000008176 lyophilized powder Substances 0.000 claims description 11
- 238000004108 freeze drying Methods 0.000 claims description 9
- 238000005238 degreasing Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000002525 ultrasonication Methods 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 7
- 238000003776 cleavage reaction Methods 0.000 abstract description 2
- 238000005336 cracking Methods 0.000 abstract description 2
- 239000003814 drug Substances 0.000 abstract description 2
- 230000007017 scission Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 28
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 description 14
- 239000000843 powder Substances 0.000 description 10
- 239000001963 growth medium Substances 0.000 description 9
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 8
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 8
- 239000006166 lysate Substances 0.000 description 8
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 description 7
- 229960003692 gamma aminobutyric acid Drugs 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 241000244203 Caenorhabditis elegans Species 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 239000006041 probiotic Substances 0.000 description 5
- 235000018291 probiotics Nutrition 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- 230000032683 aging Effects 0.000 description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 4
- 235000019341 magnesium sulphate Nutrition 0.000 description 4
- 229940099596 manganese sulfate Drugs 0.000 description 4
- 235000007079 manganese sulphate Nutrition 0.000 description 4
- 239000011702 manganese sulphate Substances 0.000 description 4
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 4
- 229920000053 polysorbate 80 Polymers 0.000 description 4
- 239000001632 sodium acetate Substances 0.000 description 4
- 235000017281 sodium acetate Nutrition 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 235000015278 beef Nutrition 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 3
- 239000001393 triammonium citrate Substances 0.000 description 3
- 235000011046 triammonium citrate Nutrition 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 235000013922 glutamic acid Nutrition 0.000 description 2
- 239000004220 glutamic acid Substances 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 230000004792 oxidative damage Effects 0.000 description 2
- 230000036542 oxidative stress Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical group [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000237903 Hirudo Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000009858 acid secretion Effects 0.000 description 1
- 230000001153 anti-wrinkle effect Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 239000006187 pill Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 239000000741 silica gel Substances 0.000 description 1
- 229910002027 silica gel Inorganic materials 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 238000004809 thin layer chromatography Methods 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
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- 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
-
- 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
-
- 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
- C12N1/06—Lysis of microorganisms
- C12N1/066—Lysis of microorganisms by physical methods
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Toxicology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The invention relates to the technical field of medicine, and particularly discloses lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of products with anti-aging effects. The lactobacillus reuteri (Limosilac tobacillus reuteri) HCLR01 has a deposit number of GDMCC No:63144. the lactobacillus reuteri cell lysate is a lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 cell lysate. Research shows that the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 has anti-aging effect; and the anti-aging effect is obviously better than that of the known lactobacillus reuteri. The lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and the cleavage product thereof have anti-aging effect; therefore, the lactobacillus reuteri (Limosilac tobacillus reuteri) HCLR01 or the cracking product thereof provided by the invention is used as an effective component and has important application value in preparing products with anti-aging effect.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparing a product with an anti-aging effect.
Background
The causative factors of body aging are complex and diverse, wherein oxidative stress and oxidative damage are important causative factors of body aging. Therefore, alleviation of oxidative stress, oxidative damage is an important measure against aging of the body. A great deal of researches show that probiotics and metabolites thereof planted in the human body have excellent antioxidant activity, and more intensive researches show that the regulation of the probiotics and metabolites thereof on intestinal microbiota can well improve the antioxidant capacity of intestinal tracts, thereby further delaying the aging of organisms.
Gamma-aminobutyric acid (gamma-aminobutyric acid) is used as a short chain fatty acid secreted by probiotics, and has the activities of reducing blood pressure, aiding sleep, enhancing liver function and the like; gamma-aminobutyric acid has anti-wrinkle and anti-aging activities in skin care products. Lactobacillus reuteri has strong adhesion capability to intestinal mucosa, can improve intestinal flora distribution, antagonizes harmful bacteria colonization, and is one of common probiotics. However, the existing known lactobacillus reuteri has smaller amount of gamma-aminobutyric acid secretion, and the anti-aging effect needs to be further improved.
Disclosure of Invention
In order to overcome at least one technical problem existing in the prior art, the invention firstly provides lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01.
The invention provides lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01, which has the preservation number of GDMCC No:63144.
the strain was deposited at the microorganism strain collection of Guangdong province (address was the microbiological institute of Guangdong province, hirudo 100 # college, 59 # building 5, guangdong province, university, city, 1 month 13).
The form of the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 is in a bent rod shape, and the tail end of the bent rod shape is round; analyzing the 16s rDNA sequence (shown as SEQ ID NO: 1), and comparing in NCBI database by using the sequencing result; thus, lactobacillus reuteri was finally identified (Limosilactobacillus reuteri).
The inventor screens out a brand new lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 in a large number of experiments, and the inventor surprisingly finds that the lactobacillus reuteri has an anti-aging effect in further researches; and the anti-aging effect is significantly better than that of the prior known lactobacillus reuteri (Limosilactobacillus reuteri).
The invention also provides a lactobacillus reuteri cell lysate, which is a lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 cell lysate.
The inventor finds that the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 cell lysate also has anti-aging effect in the research; and the anti-aging effect is also obviously better than that of the prior known lactobacillus reuteri cell lysate.
Preferably, the lactobacillus reuteri cell lysate is prepared by the following method:
(1) Mixing lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 with water, and performing ultrasonic crushing to obtain crushed liquid; freeze-drying the crushed liquid, and degreasing to obtain a freeze-dried product A;
(2) Mixing the freeze-dried product A with water, dialyzing by using a dialysis membrane with the pore diameter of 0.8-1.2 kD, and taking the freeze-dried product B of the freeze-dried dialysate to obtain the lactobacillus reuteri cell lysate.
The inventors have surprisingly found in the study that the choice of pore size of the dialysis membrane in step (2) plays a decisive role in the anti-aging effect of the prepared lactobacillus reuteri cell lysate; a great number of experimental researches show that the anti-aging effect of the lactobacillus reuteri cell lysate prepared by dialysis with the dialysis membrane with the pore diameter of 0.8-1.2 kD is greatly higher than that of the lactobacillus reuteri cell lysate prepared by dialysis with the dialysis membrane with other pore diameters.
Preferably, the time of ultrasonic crushing is 10-30 min.
Preferably, the ultrasonic power of the ultrasonic crushing is 500-1000W, and the ultrasonic frequency of the ultrasonic crushing is 20-40 KHz.
Preferably, lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 described in step (1) is lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder.
Preferably, the weight ratio of the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder to water in the step (1) is 1: 8-15.
Most preferably, the weight ratio of lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder to water in step (1) is 1:10.
preferably, in step (2) the dialysis is performed with a dialysis membrane having a pore size of 1 kD.
Preferably, the weight ratio of the lyophilized product A to water in step (2) is 1: 8-15.
Most preferably, the weight ratio of the lyophilized product a to water in step (2) is 1:10.
the invention also provides an application of the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 in preparation of products with anti-aging effect.
The invention also provides application of the lactobacillus reuteri cell lysate in preparation of products with anti-aging effect.
Preferably, the product is a food, dietary supplement, functional food, health product or pharmaceutical.
Preferably, the dosage form of the product is pill, capsule, tablet, granule or powder.
The beneficial effects are that: the invention firstly provides a brand new lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01; research shows that it has anti-aging effect; and the anti-aging effect is obviously better than that of the known lactobacillus reuteri. The lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and the cleavage product thereof have anti-aging effect; therefore, the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 or the cracking product thereof provided by the invention is used as an effective component and has important application value in preparing products with anti-aging effect.
Detailed Description
The technical scheme of the present invention will be clearly and completely described in the following examples. It will be apparent that the described embodiments are only some, but not all, embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
EXAMPLE 1 isolation and characterization of Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01
(1) Enrichment culture of strains: respectively taking fecal treatment liquid from different healthy children, adding into culture medium (peptone 8.5g/L, yeast powder 8.5g/L, glutamic acid 8.5g/L, glucose 9.5g/L, tween 80 1.2mL/L, sodium acetate 5.5g/L, dipotassium hydrogen phosphate 2.2g/L, magnesium sulfate 0.6g/L, manganese sulfate 0.4 g/L), and standing at 33deg.C for 6 hr.
(2) Qualitative detection: concentrating and qualitatively detecting the culture solution after culturing in the step (1). Thin layer chromatography, the concentrated culture solution is spotted on a silica gel precast slab, and the chromatographic solution is developed. The chromatographic liquid adopts chloroform: glacial acetic acid: water (65:25:10), 0.5% ninhydrin was added. Naturally airing after development is finished, developing at 98 ℃ for 2min, and comparing with a standard substance.
(3) Isolation of single colonies: microscopic examination is carried out on the culture solution with the gamma-aminobutyric acid generated in the step (2), after the bacterial species are judged, the culture solution is diluted and coated on a coating plate, and the coating plate culture medium contains 12g/L CaCO 3 The probiotic solid culture medium (8.8 g/L peptone, 5.5g/L beef extract powder, 8.8g/L yeast extract powder, 10.0g/L glucose, 2.2g/L dipotassium hydrogen phosphate, 2.0g/L tri-ammonium citrate, 5.5g/L sodium acetate, 0.6g/L magnesium sulfate, 0.4g/L manganese sulfate, 1.2g/L tween-80 and 16.0g/L agar) is cultured for 6 hours at 33 ℃, a plurality of colonies with dissolved calcium rings (smooth and regular edges) are picked up and inoculated on the culture medium containing glutamic acid, and the culture medium is kept at 33 ℃ for 6 hours, and the gamma-aminobutyric acid yield is qualitatively detected and compared. Selecting a strain with a rod-like shape and a round tail end and highest gamma-aminobutyric acid yield to obtain lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01.
The strain is analyzed for 16s rDNA sequence (shown as SEQ ID NO: 1), and the sequencing result is utilized for comparison in NCBI database, so that the strain is identified as lactobacillus reuteri (Limosilactobacillus reuteri).
EXAMPLE 2 preparation of Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder
(1) Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 seed liquid preparation: inoculating Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 strain into a culture medium (the culture medium comprises 8.8g/L of peptone, 5.5g/L of beef extract powder, 8.8g/L of yeast extract powder, 10.0g/L of glucose, 2.2g/L of dipotassium hydrogen phosphate, 2.0g/L of triammonium citrate, 5.5g/L of sodium acetate, 0.6g/L of magnesium sulfate, 0.4g/L of manganese sulfate, 1.2g/L of tween-80 and 16.0g/L of agar), and performing shake flask fermentation for 12h to obtain the Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 seed liquid with OD 600 more than 1.5.
(2) And (3) performing expansion culture: inoculating lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 seed liquid obtained in the step (1) into a liquid culture medium (the culture medium comprises 8.8g/L of peptone, 5.5g/L of beef extract, 8.8g/L of yeast extract, 10.0g/L of glucose, 2.2g/L of dipotassium hydrogen phosphate, 2.0g/L of tri-ammonium citrate, 5.5g/L of sodium acetate, 0.6g/L of magnesium sulfate, 0.4g/L of manganese sulfate, 1.2g/L of tween-80 and 16.0g/L of agar) in a 20L fermentation tank; at pH 6; stirring and culturing for 48h at 37 ℃; ending fermentation to obtain fermentation liquor;
(3) And (3) centrifugally collecting thalli from the fermentation liquid obtained in the step (2), adding water and the freeze-drying protective agent skimmed milk powder, uniformly mixing, and freeze-drying to obtain the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 freeze-dried powder with the viable count of 100 hundred million cfu/g.
EXAMPLE 3 preparation of Lactobacillus reuteri lysate
(1) Mixing lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder (100 hundred million cfu/g) with 10 times of water; then carrying out ultrasonic crushing for 15min under the condition that the ultrasonic power is 900W and the ultrasonic frequency is 25KHz to obtain crushing liquid; freeze-drying the crushed liquid, degreasing the crushed liquid by isopropanol to obtain a freeze-dried product A;
(2) Mixing the freeze-dried product A with 10 times of water, dialyzing by using a dialysis membrane with the pore diameter of 1kD, and taking the freeze-dried product B of the freeze-dried dialysate to obtain the lactobacillus reuteri cell lysate.
Comparative example 1 preparation of Lactobacillus reuteri lysate
(1) Mixing lactobacillus reuteri LR-G100 lyophilized powder (100 hundred million cfu/G commercially available) with 10 times of water; then carrying out ultrasonic crushing for 15min under the condition that the ultrasonic power is 900W and the ultrasonic frequency is 25KHz to obtain crushing liquid; freeze-drying the crushed liquid, degreasing the crushed liquid by isopropanol to obtain a freeze-dried product A;
(2) Mixing the freeze-dried product A with 10 times of water, dialyzing by using a dialysis membrane with the pore diameter of 1kD, and taking the freeze-dried product B of the freeze-dried dialysate to obtain the lactobacillus reuteri cell lysate.
Comparative example 2 preparation of Lactobacillus reuteri lysate
(1) Mixing lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder (100 hundred million cfu/g) with 10 times of water; then carrying out ultrasonic crushing for 15min under the condition that the ultrasonic power is 900W and the ultrasonic frequency is 25KHz to obtain crushing liquid; freeze-drying the crushed liquid, degreasing the crushed liquid by isopropanol to obtain a freeze-dried product A;
(2) Mixing the freeze-dried product A with 10 times of water, dialyzing by using a dialysis membrane with the pore diameter of 0.5kD, and taking the freeze-dried product B of the freeze-dried dialysate to obtain the lactobacillus reuteri cell lysate.
Comparative example 3 preparation of Lactobacillus reuteri lysate
(1) Mixing lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder (100 hundred million cfu/g) with 10 times of water; then carrying out ultrasonic crushing for 15min under the condition that the ultrasonic power is 900W and the ultrasonic frequency is 25KHz to obtain crushing liquid; freeze-drying the crushed liquid, degreasing the crushed liquid by isopropanol to obtain a freeze-dried product A;
(2) Mixing the freeze-dried product A with 10 times of water, dialyzing with a dialysis membrane with a pore size of 2kD, and taking the freeze-dried product B of the dialysate after freeze-drying to obtain the lactobacillus reuteri cell lysate.
Experimental example 1 anti-aging experiment
C, taking caenorhabditis elegans L4 larvae, and randomly dividing the larvae into 7 groups of 100 larvae each; wherein, the experimental group 6 and the blank control group 1; the larvae of the caenorhabditis elegans L4 of the experimental group are respectively placed on NGM flat plates containing 2mg/mL of samples to be tested for culture; placing the caenorhabditis elegans L4 larvae of the blank control group on an NGM plate without a drug to be tested for culture; marked as dead by the non-mobile caenorhabditis elegans; each group of experiments was repeated 3 times per day; calculating the average life span of each group of caenorhabditis elegans; the experimental results are shown in Table 1.
Wherein, the test sample of the experimental group 1 is lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 freeze-dried powder (100 hundred million cfu/g); the test sample of the experimental group 2 is lactobacillus reuteri LR-G100 freeze-dried powder (100 hundred million cfu/G on the market); the test sample of experimental group 3 was lactobacillus reuteri lysate prepared as described in example 3; the test sample of experiment group 4 is the lactobacillus reuteri lysate prepared according to the method described in comparative example 1; the test sample of experimental group 5 was a lactobacillus reuteri lysate prepared as described in comparative example 2; the test sample of experimental group 6 was a lactobacillus reuteri lysate prepared as described in comparative example 3.
TABLE 1 anti-aging test results
From the experimental results in Table 1, it can be seen that the average life span of C.elegans in experimental group 1 is significantly higher than that in experimental group 2. This illustrates: the brand new lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 has an anti-aging effect; and the anti-aging effect is obviously better than that of the prior known lactobacillus reuteri.
As can be seen from the experimental results in Table 1, the average life span of the caenorhabditis elegans of experimental group 3 is significantly higher than that of experimental group 4. This illustrates: the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 cell lysate also has an anti-aging effect; and the anti-aging effect is also obviously better than that of the prior known lactobacillus reuteri cell lysate.
From the experimental results in Table 1, it can be seen that the average life span of C.elegans of experimental group 3 is significantly higher than that of experimental groups 5 and 6. This illustrates: the pore size of the dialysis membrane in the step (2) plays a decisive role in the anti-aging effect of the prepared lactobacillus reuteri cell lysate; the anti-aging effect of the lactobacillus reuteri cell lysate prepared by dialysis with the dialysis membrane with the aperture of 1kD is greatly higher than that of the lactobacillus reuteri cell lysate prepared by dialysis with the dialysis membrane with other apertures; so that the prepared lactobacillus reuteri cell lysate has excellent anti-aging effect.
Claims (10)
1. Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 deposited under the accession number GDMCC No:63144.
2. a lactobacillus reuteri cell lysate, wherein the lactobacillus reuteri cell lysate is a lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 cell lysate.
3. Lactobacillus reuteri cell lysate according to claim 2, characterized in that it is prepared by the following method:
(1) Mixing lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 with water, and performing ultrasonic crushing to obtain crushed liquid; freeze-drying the crushed liquid, and degreasing to obtain a freeze-dried product A;
(2) And mixing the freeze-dried product A with water, dialyzing by using a dialysis membrane with the pore diameter of 0.8-1.2 kD, and taking a freeze-dried product B of the freeze-dried dialysate to obtain the lactobacillus reuteri cell lysate.
4. Lactobacillus reuteri cell lysate according to claim 2, characterized in that the time of the ultrasonication is 10-30 min.
5. The lactobacillus reuteri cell lysate of claim 2, wherein the ultrasonic power of the ultrasonic disruption is 500-1000 w and the ultrasonic frequency of the ultrasonic disruption is 20-40 khz.
6. Lactobacillus reuteri cell lysate according to claim 2, wherein the lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 in step (1) is a lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder.
7. The lactobacillus reuteri cell lysate of claim 6, wherein the weight ratio of lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder to water in step (1) is 1: 8-15 parts;
most preferably, the weight ratio of lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 lyophilized powder to water in step (1) is 1:10.
8. the lactobacillus reuteri cell lysate of claim 6, wherein the dialysis in step (2) is performed with a dialysis membrane having a pore size of 1 kD.
9. The lactobacillus reuteri cell lysate of claim 6, wherein the weight ratio of the lyophilized product a to water in step (2) is 1: 8-15 parts;
most preferably, the weight ratio of the lyophilized product a to water in step (2) is 1:10.
10. use of lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 as claimed in claim 1 or a lactobacillus reuteri cell lysate as claimed in any of claims 2 to 9 in the manufacture of a product having anti-ageing properties.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310530608.4A CN116355812A (en) | 2023-05-12 | 2023-05-12 | Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310530608.4A CN116355812A (en) | 2023-05-12 | 2023-05-12 | Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116355812A true CN116355812A (en) | 2023-06-30 |
Family
ID=86922843
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310530608.4A Pending CN116355812A (en) | 2023-05-12 | 2023-05-12 | Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116355812A (en) |
-
2023
- 2023-05-12 CN CN202310530608.4A patent/CN116355812A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108728382B (en) | Lactobacillus plantarum capable of reducing cholesterol and promoting intestinal tract short-chain fatty acid production and application thereof | |
| CN101068918B (en) | Lactobacillus rhamnosus with body-fat reducing activity and the foods containing them | |
| CN101260377B (en) | Animal bifidobacteria and use thereof | |
| CN108384735B (en) | Lactobacillus plantarum CCFM1019, fermented food thereof and application of lactobacillus plantarum CCFM1019 in preparation of medicines | |
| CN110564638A (en) | Lactobacillus reuteri with probiotic characteristics and application thereof | |
| CN108486000B (en) | Preparation method and application of bifidobacterium single-bacterium fermented milk | |
| CN110484459B (en) | Lactobacillus plantarum and application thereof | |
| CN114642686B (en) | Composite probiotics and its functions of delaying senility and resisting oxidation | |
| CN109481476A (en) | Application of the lactobacillus fermenti CQPC04 in the food or drug that preparation improves ulcerative colitis | |
| WO2019208151A1 (en) | Composition for type iv allergy | |
| CN115992059B (en) | Lactobacillus johnsonii for producing feruloyl esterase and application thereof in relieving ulcerative colitis | |
| KR20230154400A (en) | Lactobacillus plantarum hom3201 strain and its live bacterial preparation, preparation method and application | |
| CN114703108A (en) | Lactobacillus mucilaginosus strain and application thereof in improvement of facial redness and type I rosacea | |
| CN113337440A (en) | Lactobacillus salivarius MG-587 and application thereof | |
| CN116355812A (en) | Lactobacillus reuteri (Limosilactobacillus reuteri) HCLR01 and application thereof in preparation of anti-aging products | |
| CN112708577B (en) | Lactobacillus fermentum DALI02 with high intestinal adhesion and immunoregulation function and application thereof | |
| CN114806953A (en) | Lactobacillus gasseri with characteristic of improving type 1 diabetes | |
| TWI746955B (en) | Composition for Type I Allergy | |
| CN116590181B (en) | Lactobacillus paracasei for improving inflammatory reaction caused by microplastic pollution and application thereof | |
| CN116444605A (en) | Active peptide, active peptide composition and application thereof in preparation of anti-aging products | |
| CN117143781B (en) | Lactobacillus plantarum, mix metaplasia and preparation method and application thereof | |
| CN117467586B (en) | Lactobacillus rhamnosus with effect of delaying chronic nephrosis process | |
| CN117286057A (en) | Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof | |
| CN116445352A (en) | Lactobacillus paracasei (Lacticaseibacillus paracasei) HCLP01 and application thereof in preparation of products with antioxidant or anti-aging effects | |
| CN116987645A (en) | Lactobacillus plantarum (Lactiplantibacillus plantarum) HEPRO-185 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 |