Application of bifidobacterium breve metagen in aspects of scavenging free radicals and relieving oxidative aging

Abstract

The invention belongs to the technical field of fermentation and metagen preparations, and particularly relates to application of bifidobacterium breve metagen in the aspects of scavenging free radicals and relieving oxidative aging. According to the present invention, the oxidative free radical scavenging experiment results show that the bifidobacterium breve NKU BB 1-2 metagen has good oxidative free radical scavenging ability, and has good scavenging ability on hydroxyl free radicals, superoxide anions and DPPH free radicals; meanwhile, through component analysis, it is found that the bifidobacterium breve NKU BB 1-2 metagen contains various antioxidant and anti-aging bioactive substances such as flavonoids, indoles, amino acids, bile acids and ceramides; a cell model experiment verifies that the bifidobacterium breve NKU BB 1-2 metagen can obviously relieve oxidative damage to the cells and improve the oxidation resistance of the cells, so that oxidative aging is relieved.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of fermentation and postbiotic preparation, and particularly relates to application of Bifidobacterium breve postbiotic in removing free radicals and relieving oxidative aging. BACKGROUND

[0002] Anti-aging research has always been a hot topic in the scientific community. With the intensification of population aging, people's demand for delaying aging and maintaining health is increasingly urgent. Oxidative aging is one of the core concepts of modern aging theory. Its core view believes that continuous oxidative free radicals will be generated under life activities or special environment, such as superoxide anion, hydrogen peroxide, hydroxyl radical, etc. These oxidative free radicals will continuously attack key biological molecules in cells and have a negative impact on the key life activities of tissues and organs. In young individuals, there is usually a set of precise antioxidant defense system (such as superoxide dismutase SOD, glutathione, vitamin C / E, etc.) that can effectively remove or neutralize these oxidative free radicals to achieve a dynamic balance between oxidation and antioxidant.

[0003] Although aging is a normal life process of organisms, the occurrence of aging often leads to the tilt of oxidation-reduction homeostasis to oxidation. The weakening of the body's antioxidant capacity and the increase of endogenous oxidative free radical generation accelerate aging, and the acceleration of aging further aggravates this oxidation process. This continuous and gradually aggravated oxidative stress leads to the accumulation of biological molecule damage, such as increased DNA mutation, protein misfolding and loss of function, and lipid peroxidation damage to cell membrane structure.

[0004] Among all organ systems in the body, the intestine has become a focus area in oxidative aging research due to its unique physiological environment and function. The high metabolic rate, high exposure, and large microbial ecosystem of the intestine make it extremely susceptible to oxidative stress. Local oxidative damage in the intestine leads to barrier dysfunction and dysbiosis, triggering systemic inflammation and oxidative stress, forming a vicious cycle driving whole-body aging. Understanding the central role of the intestine in oxidative aging and studying how to maintain intestinal redox homeostasis, protect the intestinal barrier, and regulate healthy flora have become a key breakthrough and frontier research field for delaying aging and preventing and treating age-related diseases.

[0005] The International Scientific Association for Probiotics and Prebiotics (ISAPP) defines postbiotics as: “a preparation of inactivated microorganisms and / or components thereof that confer a health benefit on the host,” which must contain inactivated microbial cells or cell components (such as cell wall fragments, peptidoglycans, etc.) and have clinically proven health benefits. Compared with probiotic live bacterial preparations, postbiotics have higher stability, strong safety and more extensive application scenarios, including but not limited to regulating intestinal flora, enhancing intestinal barrier function, regulating immunity and metabolism, etc. Current research on the application of postbiotics mainly has two mainstream views: one believes that postbiotics should be regarded as a whole of a mixture of various beneficial substances, and the other believes that the components of postbiotics should be identified, and some beneficial components should be purified if necessary. However, the ISAPP consensus on postbiotics stipulates that effective postbiotics must contain inactivated microbial cells or cell components that are beneficial to the host, and microbial fermentation metabolites (fermentation supernatant) are not necessarily required, that is, according to the ISAPP consensus document, although fermentation metabolites have been experimentally verified to be beneficial to the host, they cannot be called postbiotics, but should be called cell-free supernatant, which provides scientific support for the research of postbiotics for those skilled in the art.

[0006] Bifidobacterium breve is a gram-positive anaerobic bacillus of the family Actinomycetaceae and the genus Bifidobacterium, which is mainly used in the research of fermented dairy products, dietary supplements and clinical nutrition. Bifidobacterium breve live bacteria have various effects and functions, mainly including regulating intestinal flora balance, enhancing immunity, improving digestive function and reducing the risk of intestinal diseases, etc. However, there are few studies on postbiotics of Bifidobacterium breve, especially its research on relieving oxidative aging and scavenging free radicals, which needs to be further developed. SUMMARY

[0007] The purpose of the present application is to provide the application of Bifidobacterium breve postbiotics in scavenging free radicals and relieving oxidative aging. The Bifidobacterium breve NKU BB 1-2 postbiotic preparation can effectively scavenge free radicals in the body, has the functions of antioxidant and relieving oxidative aging, and the effect is significant.

[0008] The present application provides a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, which comprises a Bifidobacterium breve NKU BB 1-2 inactivated bacterial suspension.

[0009] Preferably, the concentration of the bacterial body in the Bifidobacterium breve NKU BB 1-2 inactivated bacterial suspension is 1×(10 6 ~10 9 ) CFU / mL.

[0010] The present invention also provides a method for preparing the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution, comprising the following steps: resuspending the Bifidobacterium breve NKU BB 1-2 bacterial cells with sterile physiological saline or sterile PBS buffer to obtain a Bifidobacterium breve NKU BB 1-2 bacterial cell resuspension; The heavy suspension of Bifidobacterium breve NKU BB 1-2 was inactivated at 70-80° C. for 10-30 minutes to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0011] Preferably, the method for obtaining the bacterial cell of Bifidobacterium breve NKU BB 1-2 comprises the following steps: fermenting and culturing Bifidobacterium breve NKU BB 1-2 to obtain a fermentation culture liquid of Bifidobacterium breve NKU BB 1-2; The fermentation culture broth of Bifidobacterium breve NKU BB 1-2 is subjected to solid-liquid separation, and the precipitate is collected to obtain the bacterial cells of Bifidobacterium breve NKU BB 1-2.

[0012] The present invention also provides the use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution in food.

[0013] The present invention also provides the use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution in the preparation of antioxidant products.

[0014] Preferably, the antioxidant product includes a product having one or more functions of scavenging free radicals, delaying oxidative aging and reducing cell damage.

[0015] Preferably, the product comprises a health food.

[0016] The present invention also provides an antioxidant product, the active ingredient of which includes a Bifidobacterium breve NKU BB 1-2 postbiotic preparation; the Bifidobacterium breve NKU BB 1-2 postbiotic preparation is the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution.

[0017] Preferably, the content of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in the antioxidant product is 90.0 wt.% to 99.9 wt.%.

[0018] Beneficial effects: The present invention provides a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, comprising a heavy suspension of inactivated Bifidobacterium breve NKU BB1-2 bacteria; the deposit number of the Bifidobacterium breve NKU BB 1-2 is GDMCC No: 65956. Furthermore, the present invention also provides a method for preparing the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution, comprising the following steps: resuspending the Bifidobacterium breve NKU BB 1-2 bacteria in sterile physiological saline or sterile PBS buffer to obtain a heavy suspension of Bifidobacterium breve NKU BB 1-2 bacteria; inactivating the heavy suspension of Bifidobacterium breve NKU BB 1-2 bacteria at 70-80°C for 10-30 minutes to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation. Through oxidative free radical scavenging experiments, the present invention found that Bifidobacterium breve NKU BB 1-2 postbiotics have good oxidative free radical scavenging ability, and have good scavenging ability for hydroxyl radicals, superoxide anions and DPPH radicals; at the same time, through its component analysis, it was found that Bifidobacterium breve NKU BB 1-2 postbiotics contain flavonoids, indoles, amino acids, bile acids and ceramides and other bioactive substances with antioxidant and anti-aging properties; and cell model experiments verified that Bifidobacterium breve NKU BB 1-2 postbiotics can significantly reduce the oxidative damage suffered by cells, improve the antioxidant capacity of cells, and thus alleviate oxidative aging.

[0019] Biological deposit information Bifidobacterium breve NKU BB 1-2, biologically classified as Bifidobacterium breve , and was deposited in the Guangdong Provincial Microbial Culture Collection on March 24, 2025. The deposit address is 5th Floor, Laboratory Building, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 65956. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments are briefly introduced below.

[0021] Figure 1 This is a statistical graph of the hydroxyl radical scavenging ability of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation and the control strain postbiotic preparation in Test Example 1; Figure 2 This is a statistical graph of the superoxide anion scavenging ability of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation and the control strain postbiotic preparation in Test Example 1; Figure 3 This is a statistical graph of the DPPH radical scavenging ability of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation and the control strain postbiotic preparation in Test Example 1; Figure 4 This is a statistical chart of indole components in the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in Test Example 2; Figure 5 This is a statistical chart of amino acid components in the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in Test Example 2; Figure 6 This is a statistical chart of bile acid components in the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in Test Example 2; Figure 7 This is a statistical chart of ceramide components in the Bifidobacterium breve NKU BB 1-2 postbiotic preparation of Test Example 2; Figure 8 This is a statistical chart of flavonoid components in the Bifidobacterium breve NKU BB 1-2 postbiotic preparation of Test Example 2; Figure 9 The results show the difference in indole lactic acid and L-phenylalanine in the postbiotic preparation of Bifidobacterium breve NKU BB 1-2 under different heat treatment conditions in Test Example 2; Figure 10 The results show the difference in 3-oxo-5beta-cholic acid and dehydrolupinol in the postbiotic preparation of Bifidobacterium breve NKU BB 1-2 under different heat treatment conditions in Test Example 2; Figure 11 β-galactosidase staining images of different groups in Test Example 3; Figure 12 Statistical graph of the blue area stained with β-galactosidase in different groups in Test Example 3; Figure 13 This is a statistical chart of the results of measuring the antioxidant enzyme SOD in different groups of cells in Test Example 3; Figure 14 This is a statistical chart of the antioxidant enzyme MDA assay results in different groups of cells in Test Example 3; Figure 15 This is a statistical chart of the results of measuring the antioxidant enzyme CAT in different groups of cells in Test Example 3; Figure 16 This is a statistical graph of the GSH-PX assay results in different groups of cells in Test Example 3; exist Figures 12 to 16 “*” indicates significant difference P <0.05, “**” indicates significant difference P <0.01, “***” indicates significant difference P <0.001. DETAILED DESCRIPTION

[0022] The application provides a Bifidobacterium breve NKU BB 1-2 probiotic preparation, which comprises a Bifidobacterium breve NKU BB 1-2 inactivated bacterial suspension; the preservation number of the Bifidobacterium breve NKU BB 1-2 is GDMCC No: 65956.

[0023] The Bifidobacterium breve NKU BB 1-2 is isolated from an infant fecal sample, and in vitro antioxidant capacity evaluation experiments show that the Bifidobacterium breve NKU BB 1-2 has good oxidative free radical scavenging capacity.

[0024] As an embodiment, the concentration of the bacterial bodies in the Bifidobacterium breve NKU BB 1-2 inactivated bacterial suspension is 1× (10 6 ~10 9 ) CFU / mL; as another embodiment, the concentration of the bacterial bodies in the Bifidobacterium breve NKU BB 1-2 inactivated bacterial suspension is any concentration in the range of 1×10 6 CFU / mL, 1×10 7 CFU / mL, 1×10 8 CFU / mL, 1×10 9 CFU / mL or 1× (10 6 ~10 9 ) CFU / mL.

[0025] The application further provides a preparation method of the Bifidobacterium breve NKU BB 1-2 probiotic preparation, which comprises the following steps: resuspending Bifidobacterium breve NKU BB 1-2 bacterial bodies with sterile normal saline or sterile PBS buffer to obtain a Bifidobacterium breve NKU BB 1-2 bacterial body suspension; and inactivating the Bifidobacterium breve NKU BB 1-2 bacterial body suspension at 70-80 DEG C for 10-30 min to obtain the Bifidobacterium breve NKU BB 1-2 probiotic preparation.

[0026] As an embodiment, the present application carries out fermentation culture of Bifidobacterium breve NKU BB 1-2 to obtain fermentation culture solution of Bifidobacterium breve NKU BB 1-2. As an embodiment, the fermentation culture temperature is 36-38℃; as another embodiment, the fermentation culture temperature is 37℃. As an embodiment, the fermentation culture time is 46-50h; as another embodiment, the fermentation culture time is 47-48h; as another embodiment, the fermentation culture is carried out to the middle logarithmic growth phase of Bifidobacterium breve NKU BB 1-2; as another embodiment, the fermentation culture is carried out to the OD value of Bifidobacterium breve NKU BB 1-2 is 2.0. As an embodiment, the fermentation culture medium is TPY liquid medium. As an embodiment, the inoculation amount of Bifidobacterium breve NKU BB 1-2 is 1%-3% of the volume of the culture medium; as another embodiment, the inoculation amount of Bifidobacterium breve NKU BB 1-2 is 2% of the volume of the culture medium.

[0027] After obtaining the fermentation culture solution of Bifidobacterium breve NKU BB 1-2, as an embodiment, the present application carries out solid-liquid separation on the fermentation culture solution of Bifidobacterium breve NKU BB 1-2, takes the precipitate to obtain the Bifidobacterium breve NKU BB 1-2 cell body. As an embodiment, the solid-liquid separation method can be centrifugation; as another embodiment, the centrifugation speed can be 4000g; the centrifugation speed can be 10min.

[0028] After obtaining the Bifidobacterium breve NKU BB 1-2 cell body, as an embodiment, the present application washes the Bifidobacterium breve NKU BB 1-2 cell body for 2-3 times. As an embodiment, the present application carries out the washing with resuspension agent. As an embodiment, the resuspension agent is sterile normal saline or sterile PBS buffer.

[0029] After obtaining the Bifidobacterium breve NKU BB 1-2 cell body, the present application resuspends the Bifidobacterium breve NKU BB 1-2 cell body with sterile normal saline or sterile PBS buffer to obtain Bifidobacterium breve NKU BB 1-2 cell body resuspension. As an embodiment, the concentration of the Bifidobacterium breve NKU BB 1-2 cell body resuspension is 1×(10 6 ~10 9 )CFU / mL; as another embodiment, the concentration of the Bifidobacterium breve NKU BB 1-2 cell body resuspension of the present application is 1×10 6 CFU / mL, 1×10 7 CFU / mL, 1×10 8CFU / mL, 1×10 9 CFU / mL or 1× (10 6 ~10 9 ) CFU / mL.

[0030] After obtaining the Bifidobacterium breve NKU BB 1-2 bacterial cell resuspension, the present invention inactivates the Bifidobacterium breve NKU BB1-2 bacterial cell resuspension at 70-80°C for 10-30 minutes to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation. As one embodiment, the present invention's method of inactivating the Bifidobacterium breve NKU BB 1-2 bacterial cell resuspension after resuspension allows the Bifidobacterium breve NKU BB 1-2 bacterial cell resuspension to be evenly heated, resulting in more complete inactivation and disruption of the Bifidobacterium breve NKU BB 1-2 bacteria, thereby avoiding the problem of incomplete inactivation caused by direct heat inactivation of the bacteria. As one embodiment, the inactivation temperature can be 70-75°C; as another embodiment, the inactivation temperature can be 70°C. As one embodiment, the inactivation time can be 10-20 minutes; as another embodiment, the inactivation time can be 10 minutes. The heat treatment condition of inactivating at 70° C. for 10 minutes in the present invention can ensure that the high-abundance active ingredients are inactivated as little as possible during the heat treatment process.

[0031] The present invention also provides the use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution in food.

[0032] The present invention also provides the use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution in the preparation of antioxidant products. As one embodiment, the antioxidant product of the present invention is a product having one or more functions of scavenging free radicals, delaying oxidative aging and alleviating cell damage; as another embodiment, the antioxidant product is a product having the function of scavenging free radicals and / or delaying oxidative aging; as another embodiment, the oxidative aging can be to alleviate the oxidative aging of Caco-2 cells induced by D-galactose, and the D-galactose-induced Caco-2 cell oxidative aging model can simulate the effect of postbiotics on the alleviation of oxidative aging of intestinal epithelial cells after being ingested and entering the intestine in real application scenarios. As an embodiment, the product is a health food.

[0033] The Bifidobacterium breve NKU BB 1-2 postbiotic preparation of the present invention has good oxidative group scavenging ability and a large number of antioxidant bioactive ingredients through in vitro evaluation; the Bifidobacterium breve NKU BB 1-2 has better oxidative group scavenging ability than other Bifidobacterium breve strains; and the Bifidobacterium breve NKU BB 1-2 postbiotic preparation has been verified by cell experiments to have the effect of assisting anti-oxidation and alleviating oxidative aging.

[0034] The present invention also provides an antioxidant product, the active ingredient of which includes a Bifidobacterium breve NKU BB 1-2 postbiotic preparation; the Bifidobacterium breve NKU BB 1-2 postbiotic preparation is the Bifidobacterium breve NKU BB 1-2 postbiotic preparation described in the above technical solution or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method described in the above technical solution. As an embodiment, the content of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in the antioxidant product is 90.0wt.%~99.9wt.%. As an embodiment, the antioxidant product can be an antioxidant health food and / or food. The present invention does not specifically limit the type of excipients in the antioxidant product, and conventional excipient types in the field can be added as needed.

[0035] In order to further illustrate the present invention, the technical solution provided by the present invention is described in detail below with reference to the accompanying drawings and embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0036] In the following examples, TPY liquid medium (ingredients g / L): hydrolyzed casein 10.0 g, phytone 5.0 g, yeast powder 2.0 g, glucose 5.0 g, L-cysteine ​​0.5 g, dipotassium hydrogen phosphate 2.0 g, magnesium chloride 0.5 g, zinc sulfate 0.25 g, calcium chloride 0.15 g, ferric chloride 0.0001 g, Tween-80 1.0 g; In the following examples, unless otherwise specified, all methods are conventional methods; Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0037] Example 1 A method for preparing a Bifidobacterium breve NKU BB 1-2 postbiotic preparation comprises the following steps: Bifidobacterium breve NKU BB 1-2 was inoculated into TPY liquid medium (purchased from Qingdao Haibo Biotechnology Co., Ltd.) at an inoculation amount of 2% of the volume of the medium, and fermented at 37°C for 48h until the Bifidobacterium breve NKU BB 1-2 grew to an OD value of 2.0, to obtain a Bifidobacterium breve NKU BB 1-2 fermentation culture solution; the Bifidobacterium breve NKU BB 1-2 fermentation culture solution was centrifuged at a speed of 4000r / min for 10min, and the precipitate was taken; the precipitate was washed twice with sterile normal saline, and then resuspended with sterile normal saline to obtain a bacterial cell resuspension, and the concentration of Bifidobacterium breve NKU BB 1-2 in the bacterial cell resuspension was 1×10 9 CFU / mL.

[0038] The bacterial cell resuspension was heat-treated at 70°C for 10min to obtain a Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0039] Comparative Example 1 A method for preparing a Bifidobacterium breve NKU BB 1-2 live bacterial cell resuspension, comprising the following steps: The method of Example 1 was used to prepare a Bifidobacterium breve NKU BB 1-2 live bacterial cell resuspension, and the concentration of Bifidobacterium breve NKU BB 1-2 in the bacterial cell resuspension was 1×10 9 CFU / mL.

[0040] Comparative Example 2 A method for preparing a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, comprising the following steps: The method of Example 1 was used to prepare a bacterial cell resuspension, and the concentration of Bifidobacterium breve NKU BB 1-2 in the bacterial cell resuspension was 1×10 9 CFU / mL; The bacterial cell resuspension was subjected to ultrasonic treatment under the following conditions: 300W, 10S / 10S, 20min, to obtain a Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0041] Comparative Example 3 The method of Example 1 was used to prepare a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the bacterial cell resuspension was heat-treated at 60°C for 10min to obtain a Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0042] Comparative Example 4 The preparation method of Example 1 was used to prepare the Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the bacterial resuspension was heat-treated at 60° C. for 30 min to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0043] Example 2 The preparation method of Example 1 was used to prepare the Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the bacterial resuspension was heat-treated at 70° C. for 30 min to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0044] Example 3 The preparation method of Example 1 was used to prepare the Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the bacterial resuspension was heat-treated at 80° C. for 10 min to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0045] Example 4 The preparation method of Example 1 was used to prepare the Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the bacterial resuspension was heat-treated at 80° C. for 30 min to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

[0046] Comparative Example 5 The preparation method of Example 1 was used to prepare a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the Bifidobacterium breve NKU BB 1-2 in Example 1 was replaced with Bifidobacterium breve (B. breve) ATCC 15700 (purchased from China Industrial Culture Collection Center (CICC)).

[0047] Comparative Example 6 The preparation method of Example 1 was used to prepare a Bifidobacterium breve NKU BB 1-2 postbiotic preparation, except that the Bifidobacterium breve NKU BB 1-2 in Example 1 was replaced with Bifidobacterium breve ATCC 15701 (purchased from China Industrial Culture Collection Center (CICC)).

[0048] Test Example 1 The evaluation of the oxidative group scavenging ability of the Bifidobacterium breve NKU BB 1-2 postbiotic preparations prepared in Example 1 and Comparative Examples 1-2 and Comparative Examples 5-6 was performed as follows: 1. Hydroxyl radical scavenging ability Hydroxyl radicals act on biomolecules in the body, such as proteins, nucleic acids, and lipids, causing damage to cell structure and function, which in turn leads to metabolic disorders and disease. Hydroxyl radical scavenging ability is an important indicator of the antioxidant capacity of a substance and is widely used in the research of antioxidant health foods and / or foods and pharmaceuticals.

[0049] H2O2 / Fe 2+ The Fenton reaction generates hydroxyl radicals, which convert o-phenanthroline-Fe 2+ Fe in aqueous solution 2+ Oxidized to Fe 3 + , resulting in a decrease in the absorbance at 536 nm. The degree to which the sample inhibits the rate of decrease in absorbance at 536 nm reflects the ability of the sample to scavenge hydroxyl free radicals.

[0050] Based on the above principle, the Fenton colorimetric method was used to determine the hydroxyl radical scavenging ability of the live bacterial suspension of Bifidobacterium breve NKU BB 1-2 in Comparative Example 1 and the Bifidobacterium breve NKU BB 1-2 postbiotic preparations of Comparative Example 2, Example 1, Comparative Example 5, and Comparative Example 6. The results are as follows: Figure 1 As shown in Table 1, Figure 1 In the table, NKU BB 1-2, NKU BB 1-2 postbiotics (heat-inactivated), NKU BB 1-2 postbiotics (ultrasound), B. breve ATCC 15700, and B. breve ATCC 15701 refer to the results in Comparative Example 1, Example 1, Comparative Example 2, Comparative Example 5, and Comparative Example 6, respectively; Figure 2 and Figure 3 Same thing.

[0051] Table 1 Hydroxyl radical scavenging rate results of the live bacterial suspension of Bifidobacterium breve NKU BB 1-2 in Comparative Example 1, and the Bifidobacterium breve postbiotic preparations of Example 1, Comparative Example 2, and Comparative Examples 5-6

[0052] Depend on Figure 1As can be seen from Table 1, the hydroxyl radical scavenging abilities of the standard strains of Bifidobacterium breve postbiotics prepared under the same conditions were 11.58±1.35% and 6.62±1.08%, respectively, while the hydroxyl radical scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic was significantly higher than that of the two standard strains of Bifidobacterium breve postbiotics, and the hydroxyl radical scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic was significantly higher than that of the live strain of Bifidobacterium breve NKU BB 1-2; the hydroxyl radical scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic was not much different from that of the ultrasonically treated Bifidobacterium breve NKU BB 1-2 postbiotic.

[0053] 2. Superoxide anion scavenging ability Reactive oxygen species such as superoxide anions in organisms have immune and signal transduction functions, but when they accumulate excessively, they can damage cell membranes and biological macromolecules, leading to abnormal metabolism of cells and tissues in the body, thus causing a variety of diseases. Superoxide anions react with hydroxylamine hydrochloride to generate NO 2- , NO 2- Under the action of p-aminobenzenesulfonamide and naphthylethylenediamine hydrochloride, a red azo compound is generated with a characteristic absorption peak at 530nm. The superoxide anion content in the sample can be calculated based on the A530 value, and then the superoxide anion clearance rate can be obtained, reflecting the superoxide anion clearance ability.

[0054] Based on the above, the superoxide anion scavenging ability of the live bacterial suspension in Comparative Example 1 and the Bifidobacterium breve postbiotic preparations prepared in Comparative Example 2, Example 1 and Comparative Examples 5 to 6 was determined using the hydroxylamine method. The results are as follows: Figure 2 and as shown in Table 2.

[0055] Table 2 Superoxide anion scavenging ability results of the live bacterial suspension of Bifidobacterium breve NKU BB 1-2 in Comparative Example 1, and the Bifidobacterium breve postbiotic preparations of Example 1, Comparative Example 2, and Comparative Examples 5-6

[0056] Figure 2As can be seen from Table 2, the superoxide anion scavenging abilities of the two Bifidobacterium breve strains postbiotics prepared under the same conditions were 72.48±1.70 and 56.67±1.02, respectively. The superoxide anion scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic preparation was higher than that of the two wild-type Bifidobacterium breve strains postbiotics. The superoxide anion scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic preparation was significantly higher than that of the live Bifidobacterium breve NKU BB 1-2 strain. The superoxide anion scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic was not much different from that of the ultrasonically treated Bifidobacterium breve NKU BB 1-2 postbiotic.

[0057] 3.DPPH scavenging ability A 0.2 mM DPPH solution in anhydrous ethanol was mixed with the sample to be tested at a volume ratio of 1:1. The mixture was incubated in the dark at room temperature for 30 minutes. After centrifugation at 8000 × g for 10 minutes, the absorbance of the separated supernatant was measured at 517 nm. The DPPH free radical scavenging activity (%) was calculated as follows:

[0058] Where: AS-absorbance of sample; AB - absorbance of the blank consisting of sample and ethanol; Absorbance of the control consisting of AC-DPPH solution and deionized water This method was used to determine the live bacterial suspension in Comparative Example 1 and the Bifidobacterium breve postbiotic preparations prepared in Comparative Example 2, Example 1, and Comparative Examples 5-6. The determination results were as follows: Figure 3 and Table 3.

[0059] Table 3 DPPH clearance results of the live bacterial suspension of Bifidobacterium breve NKU BB 1-2 in Comparative Example 1, and the Bifidobacterium breve postbiotic preparations of Example 1, Comparative Example 2, and Comparative Examples 5-6

[0060] Depend on Figure 3 As can be seen from Table 3, the DPPH scavenging ability of the two standard strains of Bifidobacterium breve postbiotics prepared under the same conditions is about 70%. The DPPH scavenging ability of the Bifidobacterium breve NKU BB 1-2 postbiotic is significantly higher than that of the standard strain postbiotic. The DPPH scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic is significantly higher than that of the live strain of Bifidobacterium breve NKU BB 1-2. The DPPH scavenging ability of the heat-treated Bifidobacterium breve NKU BB 1-2 postbiotic is not much different from that of the ultrasonically treated Bifidobacterium breve NKU BB 1-2 postbiotic.

[0061] Based on the above results, the oxidative group scavenging ability of heat-inactivated Bifidobacterium breve NKU BB1-2 postbiotics was evaluated, and three main comparisons were made: 1. Comparison of the oxidative group scavenging ability of Bifidobacterium breve NKU BB 1-2 postbiotics and live Bifidobacterium breve NKU BB 1-2 bacteria; 2. The oxidative group scavenging ability of Bifidobacterium breve NKU BB 1-2 postbiotics prepared by two postbiotic preparation methods (heat inactivation and ultrasound); 3. The oxidative group scavenging ability of Bifidobacterium breve NKU BB 1-2 postbiotics and postbiotics from two other standard strains of Bifidobacterium breve. The following conclusions can be drawn: 1. Bifidobacterium breve NKU BB 1-2 postbiotics have better oxidative group scavenging ability than live Bifidobacterium breve NKU BB 1-2 bacteria; 2. The two different preparation methods of heat inactivation and ultrasound have little effect on the oxidative group scavenging ability of Bifidobacterium breve NKU BB 1-2 postbiotics. Considering the industrial production conditions and production costs, the present invention selects heat inactivation to prepare Bifidobacterium breve NKU BB 1-2 postbiotics; 3. Compared with the postbiotics of other Bifidobacterium breve strains, Bifidobacterium breve NKU BB 1-2 postbiotics show better oxidative group scavenging ability. It is reasonable to infer that the Bifidobacterium breve NKU BB 1-2 postbiotics obtained by heat inactivation treatment have excellent antioxidant capacity and have potential prebiotic properties to alleviate oxidative aging.

[0062] Test Example 2 Composition Analysis of the Bifidobacterium breve NKU BB 1-2 Postbiotic Preparation in Example 1 and Composition Analysis of the Bifidobacterium breve NKU BB 1-2 Postbiotic Preparations Prepared under Different Heat Treatment Conditions 1. Non-targeted metabolomics determination of the components of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in Example 1, the steps are as follows: (1) Sample pretreatment Take a cell (bacterial) sample and place it in an EP tube. Add 300 μL of 80% methanol aqueous solution by volume; freeze it in liquid nitrogen for 5 minutes, thaw it on ice, vortex it for 30 seconds, and sonicate it for 6 minutes; centrifuge it at 5000 rpm and 4°C for 1 minute, take the supernatant into a new centrifuge tube, and lyophilize it into a dry powder; add the corresponding 10% methanol aqueous solution according to the sample volume to dissolve it, and inject it into LC-MS for analysis.

[0063] (2) Mass spectrometry conditions The scan range was selected as m / z 100-1500. The ESI source settings were as follows: spray voltage: 3.5 kV; sheath gas flow rate: 35 psi; auxiliary gas flow rate: 10 L / min; capillary temp: 320°C; S-lens RF level: 60; auxiliary gas heater temp: 350°C; polarity: positive, negative. MS / MS secondary scans were data-dependent scans.

[0064] (3) Data processing and metabolite identification Off-line data files were converted to mzXML format using ProteoWizard. Peak extraction and quantification were performed using XCMS. Peak alignment was performed across samples using parameters such as retention time and mass-to-charge ratio. Metabolite identification was then performed by comparison to a high-quality secondary spectrum database using a 10 ppm mass tolerance and adduct ion information. Background ions were then removed using a blank sample, resulting in metabolite identification and relative quantification. Data processing was performed using the Linux operating system (CentOS version 6.6) and R and Python software. Identified metabolites were annotated using the KEGG database (https: / / www.genome.jp / kegg / pathway.html), the HMDB database (https: / / hmdb.ca / metabolites), and the LIPIDMaps database (http: / / www.lipidmaps.org / ).

[0065] 2. Analysis of postbiotic components of Bifidobacterium breve NKU BB 1-2 Non-targeted metabolic analysis revealed that the postbiotic composition of Bifidobacterium breve NKU BB 1-2 is complex. Therefore, considering the abundance, antioxidant and anti-aging properties of the components, the bioactive components that have been widely used or studied were classified and analyzed. They were mainly divided into indoles and their derivatives, amino acids, bile acids, ceramides and flavonoids. The results are as follows: Figures 4 to 8 As shown in Tables 4 to 8.

[0066] Table 4 Data table of content of indole and its derivative metabolites

[0067] Table 5 Amino acid metabolite content data

[0068] Table 6 Bile acid metabolite metabolite content data table

[0069] Table 7 Ceramide metabolite content data table

[0070] Table 8 Flavonoid metabolite content data table

[0071] By Figures 4 to 8 From Tables 4-8, among the indole and its derivatives, the proportion is relatively high for indole lactic acid (ILA) and D-tryptophan. Indole lactic acid has a significant effect on antioxidant, anti-inflammatory, immune regulation and anti-tumor, etc. It can not only scavenge free radicals, induce the expression of antioxidant enzymes by binding to the aryl hydrocarbon receptor (AhR), and activate the endogenous antioxidant pathway, but also significantly alleviate the oxidative aging process by relieving inflammation and improving immune aging. D-tryptophan can also directly neutralize free radicals, reduce lipid peroxidation and DNA oxidative damage, and protect cell integrity due to its indole structure. Among the amino acid metabolites, the proportion is relatively high for short peptides composed of multiple amino acids and phenylalanine proline. Both of them have unique biological activities in antioxidant and anti-aging. In addition to being able to directly scavenge oxidative free radicals, short peptides composed of multiple amino acids can also have a positive impact on cells and tissues through various pathways. Phenylalanine proline (Phe-Pro) is a common component of collagen repeat sequences (such as Gly-X-Y, X / Y is often Pro / Hyp), which directly participates in maintaining the stability of the collagen triple helix structure. Collagen is the core structural protein of skin, bone and connective tissue, and its integrity directly affects the anti-aging ability of the tissue. Among the bile acid metabolites, the highest proportion is 3-oxo-5beta-cholalic acid, As one of the products in the bile acid metabolic pathway, bile acid metabolites from intestinal microorganisms have been proven to be involved in the body's bile acid metabolism, and bile acid metabolism is closely related to antioxidant, immune improvement, and relief of aging. Ceramide metabolites and flavonoid metabolites are two types of bioactive substances with antioxidant and anti-aging effects that have been widely proven and widely used. Ceramide can significantly improve endothelial aging, while flavonoids generally have strong oxidative free radical scavenging capacity, which can significantly reduce the negative impact of external oxidative free radicals on tissues and organs.

[0072] 3. Analysis of differences in bioactive components of Bifidobacterium breve NKU BB 1-2 postbiotics under different heat treatment conditions in Example 1, Examples 2-4 and Comparative Examples 3-4 In order to determine the optimal heat treatment conditions for Bifidobacterium breve NKU BB 1-2, that is, to minimize the loss of active ingredients due to heat treatment while ensuring that the living cells are completely heat-inactivated, non-targeted metabolomics was used to analyze the differences in the postbiotic active ingredients of Bifidobacterium breve NKU BB 1-2 under six groups of heat treatment conditions: 60℃, 10 min (Comparative Example 3); 60℃, 30 min (Comparative Example 4); 70℃, 10 min (Example 1); 70℃, 30 min (Example 2); 80℃, 10 min (Example 3); and 80℃, 30 min (Example 4). We mainly focused on the changes in the abundance of the most abundant metabolites in the common categories of active ingredients, including indole lactic acid in indole and its derivatives, L-phenylalanine in amino acids and their derivatives, 3-oxo-5beta-cholic acid in bile acid compounds, and dehydrolupinoylphenol in flavonoids, a total of four active ingredients. The content changes of these four active ingredients under different heat treatment conditions are shown in the following table. Figures 9 and 10 As shown, the colony counts of Bifidobacterium breve NKU BB 1-2 postbiotics prepared under each heat treatment condition were performed, and the counting results are shown in Table 9.

[0073] Table 9 Postbiotic colony counts under different heat treatment conditions

[0074] Combine Figures 9 and 10 From the results in Table 9, it can be concluded that 70°C for 10 min is the most optimal heat treatment condition for Bifidobacterium breve NKU BB1-2. On this basis, extending the heat treatment time and increasing the heat treatment temperature will lead to varying degrees of loss of active ingredients; while lowering the heat treatment temperature will lead to incomplete inactivation of live bacterial cells, and live bacterial cells will exist in the postbiotics.

[0075] Test Example 3 Evaluation of the probiotic potential of Bifidobacterium breve NKU BB1-2 postbiotic in alleviating oxidative aging in Example 1 based on a D-galactose-induced Caco-2 cell oxidative aging model 1. D-galactose-induced Caco-2 cell oxidative senescence model Caco-2 cells were inoculated in 96-well plates and cultured in a CO2 incubator for 24 h to adhere. When they proliferated to about 80% under microscopic observation, the old culture medium was removed, washed twice with sterile PBS, and serum-free DMEM medium was added. After 12 h of starvation in a CO2 incubator, the old culture medium was discarded, washed twice with PBS, and serum-free medium containing D-galactose at final concentrations of 50 mM, 100 mM, 200 mM, 300 mM, 400 mM, and 500 mM was added. After 24 h of stimulation, fresh serum-free medium was added, followed by the addition of 10% CCK8 reagent. After 1 h of incubation in a CO2 incubator, the absorbance value was measured at 450 nm and calculated according to the CCK8 kit instructions of Biyun Tian Biotechnology Co., Ltd. The D-galactose concentration at which the cell viability was about 50% was taken as the induction dose to establish the D-galactose-induced Caco-2 cell senescence model. According to the results of the CCK-8 test, Caco-2 cells were induced to oxidative senescence by serum-free medium containing D-galactose at a concentration of 500 mM for 24 h to construct the Caco-2 cell oxidative senescence model.

[0076] 2. Determination of intervention dose and method of Bifidobacterium breve NKU BB 1-2 postbiotic Gradient concentrations of Bifidobacterium breve NKU BB 1-2 postbiotic preparations were prepared. The specific preparation method is as follows: the concentration of postbiotic was quantified based on the number of bacteria per milliliter of liquid before heat treatment. Sterile saline suspensions containing 10 4 , 10 5 , 10 6 , 10 7 , 10 8 , and 10 9 CFU / mL of live bacteria before heat treatment were used to prepare different concentrations of Bifidobacterium breve NKU BB 1-2 postbiotic preparations by heat treatment.

[0077] Caco-2 cells were seeded in a 96-well plate and cultured in a CO2 incubator for 24 hours to adhere to the wall. When the proliferation reached about 80% under a microscope, the old culture medium was removed, and the cells were rinsed twice with sterile PBS. Serum-free DMEM culture medium was added and the cells were starved for 12 hours in a CO2 incubator. After starvation, the old culture medium was discarded, the cells were rinsed twice with PBS, and serum-free culture medium with different concentrations of Bifidobacterium breve NCUBB 1-2 postbiotics was added. After culturing for 24 hours, the cells were removed, fresh serum-free culture medium was added, and 10% CCK8 reagent was added. After culturing in a CO2 incubator for 1 hour, the absorbance was measured at 450nm and calculated. The calculation method was based on the instructions of the CCK8 kit of Biyuntian Biotechnology Co., Ltd. The results were used to evaluate whether different concentrations of postbiotics had toxic effects on Caco-2 cells. According to the CCK-8 test results, it was found that all concentrations of postbiotics had no toxic effects on cells, so 10 was selected. 9 The postbiotics prepared from CFU / mL live bacteria were used as intervention cells in the high-dose group. 6 Postbiotics prepared from viable bacteria at a concentration of CFU / mL were used as interventional agents in the low-dose group. The intervention involved co-culturing cells with postbiotics and D-galactose for 24 hours to simulate the postbiotic intervention scenario during oxidative aging.

[0078] 3. Determination of β-galactosidase Content in Cellular Senescence Caco-2 cells were seeded in 24-well plates, and the modeling and intervention treatments were as described above. The control group was starved and cultured with fresh serum-free DMEM. After the intervention, the cell culture medium was removed, washed once with sterile PBS, and stained with 0.5 mL of β-galactosidase fixative for 15 minutes at room temperature. After washing three times with sterile PBS, 500 μL of the staining working solution was added, sealed with parafilm, and incubated overnight in a conventional incubator at 37°C. The cells were observed under a microscope, and the center of each well was observed and photographed to assess the senescence of the Caco-2 cells in each group. The results are shown in Figure 2. Figure 11 As shown, the upper and lower figures of the same group represent two repetitions of the same group, indicating that the results are non-accidental.

[0079] Figure 11 The staining pictures of Caco-2 cells in each group clearly show that under the same visual field conditions, the model group has more senescent cells stained blue-green than the control group, while in the high-dose and low-dose intervention groups administered with Bifidobacterium breve NKU BB 1-2, the blue-green color was significantly weakened, indicating a decrease in the number of senescent cells. This alleviating effect on aging is reflected in both high and low doses, and the high dose is slightly better than the low dose.

[0080] Further, Figure 12Quantitative processing of blue-green area, using Image J image processing software to measure the blue-green area in each group of cell field, it can be found that the blue-green area of the model group is significantly higher, which accurately reflects that more cell aging occurs in the model group. The intervention group of postbiotics not only reverses this oxidative aging situation, but even the high-dose intervention group shows better effect in relieving cell aging than the control group. The determination of cell aging β-galactosidase content shows that the postbiotic of Bifidobacterium breve NKU BB 1-2 has a probiotic effect of relieving Caco-2 cell aging.

[0081] 4. Cell biochemical index determination Caco-2 cells were inoculated in 12-well plates, and modeling and intervention treatment were as described above. After the intervention, the cell culture solution was aspirated, and the cells were washed twice with sterile PBS. 100 μL of cell lysis solution was added for lysis and collected in a 1.5 mL centrifuge tube. The supernatant was aspirated and subpackaged in a new centrifuge tube for determination of the contents of antioxidant enzymes SOD, MDA, CAT, and GSH-PX in the cells. The determination was performed using a commercial kit from Nanjing Jiancheng Biological Engineering Institute, and the determination method was according to the kit instructions. The test results are shown in Table 2. Figures 13-16

[0082] The test of cell SOD and MDA showed that compared with the control group, the SOD activity of the model group was greatly reduced, and the MDA content was increased. The test of cell CAT and GSH-PX showed that the activity of CAT and GSH-PX in the model group was also greatly reduced. SOD, CAT, and GSH-PX are important components of the cell's own antioxidant defense system, and play an important role in removing oxidative free radicals and reducing the negative effects of oxidative stress. MDA can reflect the degree of negative impact of oxidative stress on cells, and high MDA value often indicates stronger oxidative stress damage.

[0083] It can be seen that under the intervention of D-galactose, the cells are subjected to extremely strong oxidative stress stimulation, and are in a state of oxidation greater than antioxidant, which thus triggers the accelerated aging of the cells. The intervention of the postbiotic of Bifidobacterium breve NKU BB 1-2 significantly alleviates this negative trend, which is manifested as an increase in the activity of cell SOD, CAT, and GSH-PX, and a decrease in the content of MDA. The determination of the four oxidative damage-related indicators shows that the postbiotic of Bifidobacterium breve NKU BB 1-2 helps Caco-2 cells to remove exogenous oxidative stimulation, improves endogenous antioxidant capacity, alleviates oxidative stress on cells, and thus has a probiotic effect of relieving Caco-2 cell aging.

[0084] ​It can be concluded from the above examples that the Bifidobacterium breve NKU BB 1-2 postbiotic preparation of the present invention can effectively scavenge free radicals in the body, has the functions of anti-oxidation and alleviating oxidative aging, and the effect is significant.

[0085] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A Bifidobacterium breve ( Bifidobacterium breve ) NKU BB 1-2 postbiotic preparation, characterized in that, The invention comprises a heavy suspension of inactivated bacteria of Bifidobacterium breve NKU BB 1-2; the preservation number of the Bifidobacterium breve NKU BB 1-2 is GDMCC No: 65956.

2. The Bifidobacterium breve NKU BB 1-2 postbiotic preparation according to claim 1, characterized in that The cell concentration of the inactivated Bifidobacterium breve NKU BB 1-2 cell suspension is 1×(10 6 ~10 9 ) CFU / mL.

3. The method for preparing the Bifidobacterium breve NKU BB 1-2 postbiotic preparation according to claim 1 or 2, characterized in that: The method comprises the following steps: resuspending the cells of Bifidobacterium breve NKU BB 1-2 with sterile physiological saline or sterile PBS buffer to obtain a heavy suspension of Bifidobacterium breve NKU BB 1-2; The heavy suspension of Bifidobacterium breve NKU BB 1-2 was inactivated at 70-80° C. for 10-30 minutes to obtain the Bifidobacterium breve NKU BB 1-2 postbiotic preparation.

4. The preparation method according to claim 3, characterized in that The method for obtaining the bacterial cell of Bifidobacterium breve NKU BB 1-2 comprises the following steps: fermenting and culturing Bifidobacterium breve NKU BB 1-2 to obtain a fermentation culture liquid of Bifidobacterium breve NKU BB 1-2; The fermentation culture broth of Bifidobacterium breve NKU BB 1-2 is subjected to solid-liquid separation, and the precipitate is collected to obtain the bacterial cells of Bifidobacterium breve NKU BB 1-2.

5. Use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation according to claim 1 or 2, or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method according to claim 3 or 4 in food.

6. Use of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation according to claim 1 or 2, or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method according to claim 3 or 4, in the preparation of antioxidant products.

7. The use according to claim 6, characterized in that The antioxidant products include products with one or more functions of scavenging free radicals, delaying oxidative aging and reducing cell damage.

8. The use according to claim 6 or 7, characterized in that The products include health foods.

9. An antioxidant product, characterized in that The active ingredient includes a Bifidobacterium breve NKU BB 1-2 postbiotic preparation; the Bifidobacterium breve NKU BB 1-2 postbiotic preparation is the Bifidobacterium breve NKU BB 1-2 postbiotic preparation according to claim 1 or 2 or the Bifidobacterium breve NKU BB 1-2 postbiotic preparation prepared by the preparation method according to claim 3 or 4.

10. The antioxidant product according to claim 9, characterized in that The content of the Bifidobacterium breve NKU BB 1-2 postbiotic preparation in the antioxidant product is 90.0 wt.% to 99.9 wt.%.

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