CN117286057A - Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof - Google Patents
Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof Download PDFInfo
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- 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
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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- 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
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- 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
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Abstract
The invention relates to the technical field of medicine, and particularly discloses a bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof. The bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 has a preservation number of GDMCC 63307. The strain was deposited at the microorganism strain collection in Guangdong province (address was the microbiological institute of the Guangdong national academy of sciences of Guangzhou, mitsui, 100 th university, 59 th building, 5 th building, 3 months, 29 days 2023). The bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 has excellent antioxidant and anti-aging activities; therefore, the bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 serving as an effective component has important application value in preparing products with antioxidant and/or anti-aging effects.
Description
Technical Field
The invention relates to the technical field of probiotics, in particular to bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 and application thereof.
Background
Aging is caused by a variety of factors, of which oxidation reactions and the formation of oxidative free radicals induce and exacerbate the aging process. Oxidative free radicals can interact with genetic material, functional proteins and lipids of cells, leading to cell damage and induction, acceleration of cell senescence, and the like. And the antioxidant can neutralize the excessive oxidative free radical generated in the cells, reduce the damage to the cells, and delay the aging process.
Probiotics are a class of living bacteria that can survive in the human digestive tract and are beneficial to the host, and common probiotics are lactic acid bacteria, bifidobacteria, yeasts and the like. Research reports that probiotics can regulate intestinal flora balance, inhibit growth of harmful bacteria, and promote propagation of beneficial bacteria. Meanwhile, the probiotics have the functions of maintaining the integrity of intestinal mucosa barriers, promoting food digestion and nutrient absorption, enhancing the immune system function, inhibiting inflammatory reaction and the like. Research also shows that probiotics have a certain antioxidation effect, and can reduce oxidative damage by scavenging free radicals, improving the activity of antioxidant enzymes, inhibiting oxidative stress and the like, thereby playing the role of delaying aging. However, an effective probiotic does not perform well for all people. Therefore, the development of more species of probiotics has important application value.
Disclosure of Invention
In order to overcome at least one technical problem existing in the prior art, the invention firstly provides a bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261.
The technical scheme of the invention is as follows:
the invention firstly provides a bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 with the preservation number of GDMCC No. 63307.
The strain was deposited at the microorganism strain collection in Guangdong province (address was the microbiological institute of the Guangdong national academy of sciences of Guangzhou, mitsui, 100 th university, 59 th building, 5 th building, 3 months, 29 days 2023).
The long bifidobacterium subspecies (Bifidobacterium longum) HEPRO-261 is analyzed to obtain a 16s DRNA sequence (shown as SEQ ID NO: 1), and the 16s DRNA sequence is compared in an NCBI database by using a sequencing result, wherein the homology with the long bifidobacterium subspecies (Bifidobacterium longum) in GenBank is 99.90%; the strain was finally identified as bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261.
The inventor screens out a brand new bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 in a large number of experiments, and the inventor surprisingly finds that the bifidobacterium subspecies have the functions of antioxidation and anti-aging in further research; and the antioxidation and anti-aging effects of the lactobacillus rhamnosus are obviously superior to those of the known lactobacillus rhamnosus.
The invention also provides an application of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in preparing products with anti-aging effect.
The invention also provides an application of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in preparing products with the function of promoting in-vivo expression of resistance genes.
The invention also provides an application of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in preparing products with the function of promoting the expression of internal antioxidant enzyme genes.
The invention also provides an application of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in preparing products for promoting hsf-1 gene expression.
The invention also provides an application of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in preparing products for promoting daf-16 gene expression.
Preferably, the product is a food, a dietary supplement, a functional food, a health product or a medicament.
Preferably, the product is in the form of a pill, capsule, tablet, granule or powder.
The beneficial effects are that: the invention firstly provides a brand new bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 which has excellent antioxidant and anti-aging activities; and the antioxidation and anti-aging effects of the lactobacillus rhamnosus are obviously superior to those of the known common lactobacillus rhamnosus.
In addition, the inventors have found in the study that the Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 of the present invention also has the effect of promoting the expression of hsf-1 gene and daf-16 gene; so as to promote the expression of the internal resistance gene and the antioxidant enzyme gene; thereby playing the roles of anti-aging and antioxidation.
Because the bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 has the functions of antioxidation and anti-aging; therefore, the bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 serving as an effective component has important application value in preparing products with antioxidant and/or anti-aging effects.
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 Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261
(1) Enrichment culture: adding fecal treatment solution from healthy children into liquid culture medium (peptone 8.0g/L, yeast powder 8.0g/L, glutamic acid 8.0g/L, glucose 9.0g/L, tween 80 1.0ml/L, sodium acetate 5.0g/L, dipotassium hydrogen phosphate 2.0g/L, magnesium sulfate 0.5g/L, manganese sulfate 0.5 g/L), and standing at 33deg.C for 4 hr.
(2) And (3) microscopic examination is carried out on the culture solution in the step (1), after the bacterial species are judged, the culture solution is diluted, and the bacteria-containing culture solution is coated on a coating plate for culture, different bacterial colonies are picked up for inoculating different culture mediums, and the culture is enlarged. Probiotic solid culture medium (peptone 8.0g/L, beef extract powder 5.0g/L, yeast extract powder 8.0g/L, glucose 10.0g/L, dipotassium hydrogen phosphate 2.0g/L, triammonium citrate 2.0g/L, sodium acetate 5.0g/L, magnesium sulfate 0.5g/L, manganese sulfate 0.5g/L, tween-80.0 g/L, agar 16.0 g/L) and culturing at 33 ℃ for 4h.
(3) Separation of probiotics with antioxidant and anti-aging activities: screening probiotics with antioxidant and anti-aging activities based on the constructed caenorhabditis elegans oxidation and aging model; screening one of the strains with excellent antioxidant and anti-aging activities; the 16s DRNA sequence (shown as SEQ ID NO: 1) is analyzed, and the sequence is utilized to carry out comparison in NCBI database, so that the homology with bifidobacterium longum subspecies longum (Bifidobacterium longum) in GenBank is 99.95 percent; the strain was finally identified as bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261.
EXAMPLE 2 preparation of HEPRO-261 lyophilized powder of Bifidobacterium longum subspecies longum (Bifidobacterium longum)
(1) Preparation of Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 seed solution: inoculating HCLP01 strain into culture medium (the culture medium comprises peptone 11.0g/L, yeast powder 11.0g/L, glucose 11.0g/L, tween 80 1.2ml/L, sodium acetate 6.0g/L, dipotassium hydrogen phosphate 2.5g/L, magnesium sulfate 0.7g/L, manganese sulfate 0.5g/L, glutamic acid 11.0 g/L), shaking and fermenting for 12h to obtain Bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 seed solution with OD 600 > 1.0.
(2) And (3) performing expansion culture: inoculating the Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 seed solution obtained in the step (1) into a liquid culture medium (10L, wherein the culture medium comprises 11.0g/L of peptone, 11.0g/L of yeast powder, 11.0g/L of glucose, 1.2mL/L of Tween 80, 6.0g/L of sodium acetate, 2.5g/L of dipotassium hydrogen phosphate, 0.7g/L of magnesium sulfate, 0.5g/L of manganese sulfate and 11.0g/L of glutamic acid) in a 10L fermentation tank; stirring and culturing for 24 hours, and ending fermentation to obtain fermentation liquor;
(3) And (3) centrifugally collecting thalli from the fermentation liquid obtained in the step (2), adding water and freeze-drying protective agent skimmed milk powder, uniformly mixing, and freeze-drying to obtain the bifidobacterium longum subspecies (Bifidobacterium longum) HEPRO-261 freeze-dried powder with the viable count of 10 hundred million cfu/g.
Effect example 1
The test samples were: HEPRO-261 (viable count: 10 hundred million cfu/g) of Bifidobacterium longum subspecies (Bifidobacterium longum) prepared as described in example 2 and a commercially available Lactobacillus rhamnosus LGG lyophilized powder (viable count: 100 hundred million cfu/g) were used.
Age-synchronized C.elegans at stage L1 were grown to stage L4. L4 caenorhabditis elegans (n=100) were then transferred to fresh NGM plates containing 1mg/mL of active (bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261, commercially available lactobacillus rhamnosus LGG lyophilisate) and 50 μm 5-fluoro-2-deoxyuridine (FUDR, oviposition prevention) and incubated at 20 ℃. Caenorhabditis elegans were transferred every other day to fresh NGM plates containing the active. The survival of caenorhabditis elegans was counted under the microscope daily until all caenorhabditis elegans died, and the experiment was repeated 3 times independently. The test results are shown in Table 1.
TABLE 1 anti-aging test results
As can be seen from the experimental data in Table 1, the average life span of the freeze-dried powder of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 with the viable count of 10 hundred million cfu/g is obviously longer than that of the freeze-dried powder of the commercial lactobacillus rhamnosus LGG with the viable count of 100 hundred million cfu/g. This illustrates: the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 has excellent anti-aging activity; and the anti-aging effect is obviously superior to that of the known common lactobacillus rhamnosus.
Effect example 2
Since caenorhabditis elegans decline with age both behaviorally and physiologically in a manner similar to humans. Furthermore, caenorhabditis elegans has up to 80% homology with human genes at the gene and protein level, and conserved proteins involved in senescence; thus, caenorhabditis elegans is a very popular animal model for use in anti-aging evaluation methods.
The test samples were: HEPRO-261 freeze-dried powder (viable count is 10 hundred million cfu/g) of bifidobacterium longum subspecies longum (Bifidobacterium longum) and LGG freeze-dried powder (viable count is 100 hundred million cfu/g) of lactobacillus rhamnosus are prepared according to the method described in the example 2.
Age-synchronized C.elegans at stage L1 were grown to stage L4. L4 caenorhabditis elegans were then transferred (n=100) to fresh NGM plates and after 12h co-cultivation with 10mM paraquat. Caenorhabditis elegans were collected and transferred to fresh NGM containing 1mg/mL of active substance (Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 lyophilized powder, commercially available Lactobacillus rhamnosus LGG lyophilized powder) and 50. Mu.M 5-fluoro-2-deoxyuridine (FUDR, oviposition prevention), and after incubation at 20℃for 24 hours, caenorhabditis elegans was collected, total RNA was extracted with Trizol reagent, and 2 was used ΔΔCT Analyzing the gene expression data of daf-16 and hsf-1 by the method; the test results are shown in Table 2.
TABLE 2 nematode resistance Gene expression test results
As can be seen from the experimental results in Table 2, the freeze-dried powder of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 with the viable count of 10 hundred million cfu/g can significantly up-regulate the gene expression of daf-16 and hsf-1; and the expression promotion effect of daf-16 and hsf-1 genes is significantly higher than that of the known common lactobacillus rhamnosus.
The foregoing disclosure is illustrative of the preferred embodiments of the present invention, and is not to be construed as limiting the scope of the invention, as it is understood by those skilled in the art that all or part of the above-described embodiments may be practiced with equivalents thereof, which fall within the scope of the invention as defined by the appended claims.
Claims (8)
1. A bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261, deposited under the accession number GDMCC 63312.
2. Use of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 as claimed in claim 1 in the manufacture of a product having anti-ageing properties.
3. Use of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 as claimed in claim 1 in the manufacture of a product having an effect of promoting expression of a resistance gene in vivo.
4. Use of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 as claimed in claim 1 in the manufacture of a product having an effect of promoting expression of an internal antioxidant enzyme gene.
5. Use of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 as claimed in claim 1 in the manufacture of a product having enhanced hsf-1 gene expression.
6. Use of the bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 in accordance with claim 1 for the manufacture of a product having the ability to promote expression of the daf-16 gene.
7. The use according to any one of claims 2 to 4, wherein the product is a food, a dietary supplement, a functional food, a health product or a medicament.
8. The use according to any one of claims 2 to 4, wherein the product is in the form of a pill, capsule, tablet, granule or powder.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311095118.2A CN117286057A (en) | 2023-08-29 | 2023-08-29 | Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311095118.2A CN117286057A (en) | 2023-08-29 | 2023-08-29 | Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof |
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| Publication Number | Publication Date |
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| CN117286057A true CN117286057A (en) | 2023-12-26 |
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| CN202311095118.2A Pending CN117286057A (en) | 2023-08-29 | 2023-08-29 | Bifidobacterium longum subspecies longum (Bifidobacterium longum) HEPRO-261 and application thereof |
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| Country | Link |
|---|---|
| CN (1) | CN117286057A (en) |
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