WO2018218693A1

WO2018218693A1 - Lactobacillus acidophilus having cephalosporin resistance and high expression of sir2 protein, and application thereof

Info

Publication number: WO2018218693A1
Application number: PCT/CN2017/087284
Authority: WO
Inventors: 孙晗笑; 郭晴; 利时雨; 刘梦鸽; 陈露平; 黄金群
Original assignee: 广州弘宝元生物科技有限公司
Priority date: 2017-05-28
Filing date: 2017-06-06
Publication date: 2018-12-06
Also published as: CN107227269A; CN107227269B

Abstract

Provided are a Lactobacillus acidophilus LA17 having cephalosporin resistance and high expression of Sir2 protein, and application thereof in preparing foods or medicines for anti-aging, enhancing immune function, or regulating intestinal microbiota.

Description

Lactobacillus acidophilus with cephalosporin resistance and high expression of Sir2 protein and application thereof

Technical field

The present invention belongs to the field of screening and application of new strains, and more particularly, to a Lactobacillus acidophilus LA17 having cephalosporin resistance and high expression of Sir2 protein and its use in related foods or medicines.

Background technique

Lactobacillus refers to a kind of Gram-positive bacteria that uses sugar as raw material and consumes glucose to produce lactic acid. It does not form spores, is negative for touch enzyme, lacks cytochrome, and has strict nutritional requirements. In addition to adding appropriate amount of water and carbon source during cultivation. In addition to nitrogen and inorganic salts, vitamins, amino acids and peptides are required to be added. Acid resistance, pH optimum is 5.5-6.0, optimum temperature is 30-40 °C, facultative anaerobic, low G+C content in DNA. At 55%, it is widely distributed in nature and is one of the important probiotic flora in animals and human intestines and vagina. Lactobacillus is the largest genus of lactic acid bacteria. At present, more than 40 kinds of lactobacilli have been isolated from mammalian feces, vaginal secretions, cheese and sauerkraut, including Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus bulgaricus and Lactobacillus reuteri. As a probiotic, Lactobacillus can exhibit certain probiotic functions in the human body, such as preventing the invasion and colonization of pathogenic bacteria into the intestinal tract, inhibiting the growth of pathogenic bacteria, resisting infection and maintaining the micro-ecological balance of the intestinal tract. Lactobacillus is not only used to enhance the immune, nutrition, growth and stimulation of patients, but also widely used in animal husbandry, food processing and other industries. It can be seen that Lactobacillus plays a very important role in the health and life of the human body.

Lactobacillus, as a microorganism of generalized as safe (GRAS), is basically pathogenic and has a wide range of vital physiological functions, especially to play a mutually beneficial symbiosis with the host and maintain the host bacteria. The probiotic effect of group balance. Therefore, the probiotic benefits of Lactobacillus as a probiotic are not to be underestimated, and these effects are constantly being explored and deepened.

The Sir2 (silence information regulator) gene is a class of NAD+-dependent histone/non-histone deacetylases that are highly conserved from bacteria to higher eukaryotes. The Sir2-related protein encoded by the Sir2 family gene is collectively named Sirtuin. Sir2 protein plays an important role in the regulation of physiological activities such as cell survival, apoptosis and senescence. There have been no reports of Lactobacillus acidophilus that highly express the Sir2 protein.

Summary of the invention

The present invention further studies Sir2 protein and Lactobacillus acidophilus by screening for Lactobacillus acidophilus having cephalosporin resistance, and screening for Lactobacillus acidophilus which expresses Sir2 protein by immunoblotting to make it in disease treatment or related foods. And health products have a role to play.

In order to achieve the above object, the present invention screens a Lactobacillus acidophilus LA17 having cephalosporin resistance and high expression of Sir2 protein, and is deposited in the Guangdong Provincial Collection of Microorganisms and Cultures, with the preservation number GDMCC No. 60192, the preservation address. It is the 5th floor of Building 59, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province. The deposit date is May 24, 2017.

Lactobacillus acidophilus LA17, which has cephalosporin resistance and high expression of Sir2 protein, can be used for preparing foods or medicines for anti-aging, enhancing immune function, antibacterial or regulating intestinal flora.

In order to achieve the above object, the present invention also provides an anti-aging food comprising an effective amount of the cephalosporin-resistant and high-expressing Sir2 protein-producing Lactobacillus acidophilus LA17 and/or its fungus as an active ingredient Body protein component.

In order to achieve the above object, the present invention also provides an anti-aging drug comprising an effective amount of the cephalosporin-resistant and high-expressing Sir2 protein-producing Lactobacillus acidophilus LA17 and/or its fungus as an active ingredient Body protein component, and a pharmaceutically acceptable carrier.

In order to achieve the above object, the present invention also provides an immune-enhancing food comprising an effective amount of the cephalosporin-resistant and high-expressing Sir2 protein-producing Lactobacillus acidophilus LA17 and/or as an active ingredient. Its bacterial protein component.

In order to achieve the above object, the present invention also provides an immunostimulating drug comprising an effective amount of the cephalosporin-resistant and high-expressing Sir2 protein-producing Lactobacillus acidophilus LA17 and/or as an active ingredient. Its bacterial protein component, and a pharmaceutically acceptable carrier.

In order to achieve the above object, the present invention also provides an antibacterial agent comprising an effective amount of the cephalosporin-resistant and high-expressing Sir2 protein-producing Lactobacillus acidophilus LA17 and/or a bacterial cell thereof as an active ingredient a protein component, and a pharmaceutically acceptable carrier.

In order to achieve the above object, the present invention also provides a food for regulating intestinal flora comprising an effective amount of the Lactobacillus acidophilus Lactobacillus having cephalosporin resistance and high expression of Sir2 protein according to claim 1 as an active ingredient. Acidophilus LA17 and / or its bacterial protein component. The food is particularly suitable for the regulation of intestinal flora disturbances such as diarrhea caused by the use of antibiotics, and restores the balance of the intestinal flora.

In order to achieve the above object, the present invention also provides a medicine for regulating intestinal flora, which comprises an effective amount of the Lactobacillus acidophilus Lactobacillus having cephalosporin resistance and high expression of Sir2 protein according to claim 1 as an active ingredient. Acidophilus LA17 and/or its bacterial protein component, and a pharmaceutically acceptable carrier. The drug is especially suitable for the regulation of intestinal flora disturbances such as diarrhea caused by the use of antibiotics, and restores the balance of intestinal flora.

Compared with the prior art, the present inventors have found that Lactobacillus acidophilus LA17 having cephalosporin resistance and high expression of Sir2 protein has certain antibacterial activity, and has strong cephalosporin resistance, which can significantly improve organs. The MAD content reduces the activity of SOD, GSH-Px, NO and NOS. Therefore, the Lactobacillus acidophilus LA17 having the cephalosporin resistance and high expression of the Sir2 protein can improve the body's antioxidant capacity and remove the aging body. Excessive free radicals inhibit the peroxidation process of the body, tissues and cells, and ultimately play a role in delaying aging and enhancing immunity. They can be used to prepare related foods or medicines and have good application value.

DRAWINGS

Figure 1 shows the results of identification of the sir2 gene of Lactobacillus acidophilus in the present invention.

2 is an electrophoresis pattern of Lactobacillus acidophilus protein of the present invention.

Figure 3 is a result of Western blot analysis of the Lactobacillus acidophilus Sir2 protein of the present invention.

detailed description

The present invention will be further described in detail below with reference to the embodiments.

Example 1

1. Probiotic screening

Collect 1.0g of 3 normal human feces samples, expand and culture with LB medium, dilute to 10 ^-8 total dilutions with distilled water, take 1mL, and apply to MRS separation solid culture containing 8～12g/L calcium carbonate. Base and modified MRS were separated on solid medium, placed in an anaerobic incubator, and anaerobic cultured at 37 ° C for 48 h. After the completion of the culture, the plate was taken out and the colonies with visible calcium lysate or blue and light blue were selected and transferred to the MRS enrichment broth medium for 48 h at 37 ° C, and placed in a refrigerator at 4 ° C for storage. spare.

2. Morphological observation of the strain.

The pure culture of the isolated and purified strain was inoculated into the enrichment liquid medium, incubated at 37 ° C for 24 h, and after Gram staining, the cell morphology was observed under a light microscope, and a microscope with a digital imaging system was used. Select a suitable field of view for photographic recording.

3. Molecular biology - 16S rDNA identification:

16S rDNA full-length universal primer: Primer 1 is shown in SEQ ID NO: 1; Primer 2 is shown in SEQ ID NO: 2.

PCR reaction system (25 μL): primer 1/1.0 μL; primer 2/1.0 μL; 10×PCR Buffer/2.5 μL; dNTP mix/2.0 μL; Taq enzyme/0.3 μL; DNA template/1.0 μL; ultrapure water/25 μL .

The PCR reaction conditions were: 94 ° C - 4 min; then 94 ° C - 30 s, 55 ° C - 40 s, 72 ° C - 90 s cycle 30 times, and finally 72 ° C - 10 min.

The template was replaced with sterile deionized water as a negative control. After the completion of the amplification, 3.0 μL of the amplified product was subjected to agarose gel electrophoresis. After the electrophoresis is finished, the tape to be tested is sent to Shanghai Biotech for sequence determination. The sequence results of the assay strains were compared with the known 16S rDNA sequences in NCBI by Blast to obtain the closest strain to determine the species of the bacteria isolated from the experiment.

4. Determination of antibacterial activity

The test bacteria were the above-obtained bacteria, and the control strain was Lactobacillus rhamnosus, and 100 μL of each strain was subjected to activation and resuscitation in 100 mL of lactic acid bacteria MRS enrichment broth medium. The indicator bacteria were Escherichia coli ATCC11105, Clostridium difficile NY-5, Staphylococcus aureus NT-12.

5. Drug sensitivity test

Select 10 antibacterial drug sensitive papers: cefepime (FEP, 20μg), cefotaxime (CTX, 20μg), rifampicin (Rf, 10μg), ampicillin (AM, 10μg), tetracycline (TE, 20μg ), chloramphenicol (CHL, 20 μg), ciprofloxacin (CIP, 10 μg), amikacin (AK, 20 μg), gentamicin (GM, 10 μg), trimethoprim (TMP, 10 μg) ), all purchased from Guangzhou Yikang Biotechnology Co., Ltd. (Oxoid).

Susceptibility testing was performed according to the KB method promulgated by the American Association of Clinical Laboratory Standards. First gripping respectively containing different antibiotic susceptibility paper with sterile forceps, the plates are attached to the inoculated test bacteria (200 L bacterial suspension test bacteria number of bacteria 3.0 × 10 ⁸ CFU / mL, were added In a lactic acid bacteria MRS enrichment broth agar medium with a temperature of about 50 ° C, mix well and pour the plate. Approximately 5 sheets of susceptibility paper are attached to each plate, and the distance between the sheets is approximately equal and marked. The anaerobic culture was carried out at 37 ° C overnight, and the inhibition of the plate was observed. The size of the inhibition zone was measured by a vernier caliper and recorded.

6. Identification of sir2 gene

Using the genomic DNA of the test bacteria as a template, the Lactobacillus acidophilus sir2 primer: the upstream primer is shown in SEQ ID NO: 3, and the downstream primer is shown in SEQ ID NO: 4.

PCR reaction system: template - 1 μL; upstream primer - 0.5 μL; downstream primer - 0.5 μL; dNTP (10 mM) - 0.5 μL; LA Taq (5 U / μL) - 0.5 μL; 10 × PCR buffer - 2.5 μL; - 19.5 μL.

7.Tricine-SDS-PAGE

A. Protein peptide rough

(a) 100 ml of the bacterial solution after culturing overnight in DMEM buffer medium, centrifuged at 10,000 rpm for 10 min, and separately collected the bacterial cell pellet and the supernatant;

(b) The bacterial cell pellet was transferred to the broken wall tube, mechanically broken, with a coefficient of 4.0, the time was 1 min, repeated 6 times, added with appropriate amount of deionized water, centrifuged at 10,000 rpm for 10 min, the precipitate was removed, and the supernatant was collected;

(c) Wash the filter membrane module 3 times with sterile distilled water, wash the protective solution, drain the cleaning solution, install the 10 kDa hollow fiber filter membrane, install the membrane module, and rinse the module 3 times with PBS buffer;

(d) passing the supernatant collected after centrifugation through a filter membrane module, the pressure is 0.15 MPa, the temperature is 37 ° C, and the time is 2 h, and the obtained residue is dissolved in an appropriate amount of sterile water, and the filtration is repeated twice;

(e) Take the filtrate, add solid ammonium sulfate to a saturation of 40%, let stand for 30 min, remove the precipitate by centrifugation, and the supernatant obtained after centrifugation continues to add solid ammonium sulfate to a saturation of 60%, and static at 0 °C. After 3 hours, centrifuge at 12000 rpm for 10 min, discard the supernatant, and collect the precipitate.

(f) Washing the precipitate twice with 0.1 M ammonium acetate in methanol, rinsing three times with 80% pre-cooled acetone solution, adding 100% frozen acetone once, drying at low temperature for 30 min, adding appropriate amount of distilled water to dissolve the dried product. spare.

B. Electrophoresis

(a) firstly inject the separation glue, and after the separation gel is solidified for 45 minutes, the interlayer glue is injected again, and the interlayer glue is solidified for about 45 minutes, and finally the concentrated glue is added and solidified for 45 minutes;

(b) Add the positive buffer to the bottom of the electrophoresis tank, put the prepared rubber plate into the electrophoresis tank together with the gel making device, add the negative buffer between the rubber layers, and place the whole electrophoresis device on ice with 60V. The voltage is first electrophoresed for 30 min;

(c) Take a small amount of the isolated protein sample, dilute to 5 ml with sample buffer, and start the sample electrophoresis. When the dye enters the separation gel from the interlayer glue, the voltage is applied to 120v, and the electrophoresis is continued until the dye reaches the top of the separation gel and stops. Electrophoresis

(d) Take out the rubber plate in the electrophoresis tank, immediately place it in the fixing solution, soak for 1.5h, and then take out the rubber sheet. Soak for 1.5h in the dyeing solution at 45 ° C, then wash the rubber plate twice with sterile distilled water, and finally wash it with the changing decolorizing solution, and stop the washing when the obvious band and clear background are seen;

(e) After elution, imaging with a gel imager, observing the expression of the protein of interest, and photographing and storing.

8.Western blotting

(1) Glue: Dispose 12% of the separation gel and inject it into the gap of the glass plate to 1.5cm from the upper edge, add the appropriate amount of 75% ethanol; to solidify, pour off the upper liquid, inject 5% concentrated glue, insert the comb, naturally Dry.

(2) Loading: The protein sample was bathed at 100 ° C for 3 min, and the newly configured electrophoresis solution was poured into the electrophoresis tank. 8 μl and 5 μl of protein maker were added to the two lanes, and the volume was made up to 10 μl with 1× protein loading buffer. Add 10 μl samples to each of the remaining lanes and run at 50 V constant pressure.

(3) Transfer film: After electrophoresis, cut 11cm×8cm filter paper and 0.22μm PVDF film of appropriate size and activate with methanol for 5min, according to “sponge-6 layer filter paper-gel-PVDF film-6 layer filter paper-sponge "Assemble the transfer interlayer, the splint was assembled and transferred to the transfer tank, and transferred at a constant current of 200 mA for 60 min.

(4) Incubation of antibody: After the membrane is transferred, the membrane is washed for 5 min in TBS, 5% skim milk powder is blocked for 1 h, and TBST solution is washed 3 times for 5 min each time. Add anti-human anti-human SIRT3 polyclonal antibody dilution 4 °C Incubate overnight; TBST solution was washed 3 times for 5 min each time, and the anti-diluted rabbit anti-diluted solution was incubated for 1 h at room temperature.

(5) Luminescence detection: TBST liquid washing membrane, incubated with luminescent liquid for 3 min, blotting the luminescent liquid with filter paper, placing the film and PVDF film into the tableting box for 1 min, taking out the film fixing for 30 s, washing with water, drying, scanning.

Experimental results:

1. The test results of the growth of the cells are shown in Table 1.

Table 1 results of bacterial growth test

2. 16S rDNA identification results

Enter the NCBI Nucleic Acid Database http://blast.ncbi.nlm.nih.gov/Blast.cgi, enter the 16S rDNA sequence of the strain to be tested, and click Blast to enter the alignment.

The results of the comparison showed that the 16S rDNA sequence of the two strains was 98% similar to the 16S rDNA sequences of other strains of Lactobacillus acidophilus, and it was initially identified as Lactobacillus acidophilus, which was named Lactobacillus acidophilus. LA15, Lactobacillus acidophilus LA17.

3. Determination of antibacterial activity

The test results of the antibacterial activity, as shown in Table 2, both strains of Lactobacillus acidophilus have certain antibacterial ability.

Table 2 Test results of L. acidophilus bacteriostatic activity

4. Susceptibility test, the results are shown in Table 3. The two strains of Lactobacillus acidophilus screened have strong cephalosporins Resistance.

Table 3 Results of drug sensitivity test of L. acidophilus

Note: S-sensitive; R-resistant; I-moderate drug resistance.

5.Sir2 gene PCR identification results

As shown in Figure 1, two Lactobacillus acidophilus sir2 gene bands were amplified at 720 bp, respectively.

6.Tricine-SDS-PAGE results

As shown in Figure 2, the two Lactobacillus acidophilus Sir2 proteins each had a bright band at 2.7 KD.

7.Western blotting

As shown in Figure 3, the expression of Sir2 protein in L. acidophilus LA17 was the highest. Therefore, L. acidophilus LA17 with cephalosporin resistance and high expression of the Sir2 protein was successfully screened.

Example 2

In this example, the subacute aging mice induced by D-galactose were administered to the aging mouse model by L. acidophilus LA17 (hereinafter referred to as LA17), and the index and biochemical indexes of various organs of the mouse were measured and compared. To explore the role of LA17.

Animal grouping

40 Kunming mice, half male and half female, weighing (20±2) g, were provided by the Experimental Animal Center of Jinan University. After adaptive breeding of 1W, they were randomly divided into four groups of 10 each. The first group was a blank control group, the second group was a model control group, and the third group was a Lactobacillus acidophilus group.

2. Drugs and reagents

LA17 was obtained in Example 1; D-galactose was purchased from Beijing Dingguo Biological Co., Ltd., and SOD, MDA, GSH-Px, NO, and NOS kits were purchased from Nanjing Jiancheng Bioengineering Research Institute.

3. Model establishment and drug administration plan

The model control group and the administration group received daily subcutaneous injection of D-galactose 0.15 mg/g body weight (prepared in physiological saline), and the control group was injected with the same amount of normal saline. Gastric LA17 solution (concentration to 10 ¹⁰ , daily intragastric administration of 1 mL), the blank control group and the model control group were given the same amount of normal saline for 40 days.

4. Test indicators

After continuous treatment for 40 days, the animals in each group were weighed, sacrificed by cervical dislocation, and the brain, thymus, spleen, liver and kidney were taken out. The residual blood was washed with 0.9% physiological saline and accurately weighed to calculate the organ index. The brain tissue used for sectioning was fixed in 10% formaldehyde, and the remaining organ tissues were wrapped with tin foil paper and stored in an ultra-low temperature refrigerator at -80 °C for use.

5. Determination of each organ index

After accurately weighing the whole brain, thymus, and spleen wet weight of the mouse, the index of each organ (mg/g) [thym weight (mg) / body weight (g)] was calculated.

6. Determination of biochemical indicators in brain tissue

The frozen brain tissue was placed in a glass tube for use in a tissue homogenizer, and 9 volumes of pre-cooled physiological saline (distilled water) was added thereto, and after thorough grinding, a 10% brain tissue homogenate was prepared. The content of total protein in brain homogenate was determined by Coomassie brilliant blue method; the content of MDA was determined by thiobarbituric acid reaction method; the activity of GSH-Px was detected by enzymatic method; the specific detection method of NO by nitrate reduction method was referred to the kit manual.

7. Statistical processing

All data were expressed as x±s. The comparison between groups was performed by SPSS13.0 statistical analysis software for one-way analysis of variance.

Table 4 Effect of experimental group on organ index of aging mice

Table 5 Effect of experimental group on biochemical indicators in brain tissue

result:

1. The effect of LA17 on the organ index of aging mice (see Table 4). Compared with the blank control group, the brain, thymus, spleen, liver and kidney indexes of the model group were decreased (P<0.05), and the index of each organ in the experimental group was significantly increased.

The weight of tissues and organs, especially the weight changes of vital organs such as brain, thymus, spleen, liver and kidney are important indicators reflecting the degree of aging of organisms. The most obvious immune system decay is the thymus and spleen. The reduction of thymus and spleen weight, atrophy, and functional decline can reduce immune function, leading to an increase in the incidence of malignant diseases in elderly animals (human) and accelerate the aging process. The results showed that D-galactose-induced subacute aging cerebellum, thymus, spleen, liver and kidney index were significantly decreased. LA17 treatment significantly increased the weight of brain, thymus, spleen, liver and kidney in aging mice, indicating that LA17 can effectively fight against The aging of the mouse brain, maintaining the body's immune function, and improving disease resistance, has a positive significance for delaying aging and enhancing immunity.

2. The effect of LA17 on SOD, MAD, GSH-Px, NO and NOS in aging mouse tissues (see Table 5). Compared with the blank control group, the MAD content in the brain tissue of the model group was significantly increased, the difference was statistically significant (P<0.01), and the activities of SOD, GSH-Px, NO and NOS were significantly decreased (P<0.01). The difference was statistical. Significance (P<0.01).

The organism has an antioxidant system in which SOD is one of the most critical enzymes. This enzyme can scavenge superoxide anion free radicals and protect cells from damage. Its activity reflects the antioxidant capacity of the body. Studies by the American Bardimo Medical Research Center have shown that there is a significant positive correlation between SOD content and life span in mammals. The results of this experiment showed that LA17 can significantly enhance the SOD activity of brain tissue induced by D-galactose in aging mice, improve the ability of brain tissue to scavenge free radicals, and block free radical response.

Free radicals are produced by normal metabolism of the body, but excessive free radicals can invade cells, especially unsaturated fatty acids in brain tissue, causing lipid peroxidation, forming metabolite MDA, which in turn forms lipofuscin, through proteins and nucleic acids. Cross-linking of biological macromolecules, destroying the function of normal cells. Therefore, the MDA content often reflects the degree of lipid peroxidation in the body, indirectly reflecting the degree of cell damage, and is an important indicator of aging.

NO is endogenously synthesized by NOS using L-arginine. It is a small molecular substance with a wide range of biological functions. It is soluble in water and lipids, and can rapidly diffuse to adjacent cells through paracrine, and intracellular ornithine ring. The heme gene on the enzyme activates the enzyme, and ornithine cyclase catalyzes the production of ornithine (cGMP). As a second messenger, cGMP causes a series of biological effects through signal cascade amplification. The main role of NO in the nervous system is to mediate the response of neurons to excitatory amino acids and enhance the body's learning and memory ability. Its content is reduced and aging and many senile diseases such as senile dementia, hypertension, atherosclerosis. It is related to the occurrence. NOS is a rate-limiting enzyme for NO synthesis. Neurons containing NOS are present in many parts of the central nervous system, including the cerebral cortex, hippocampus, and hypothalamus. NO plays an important role in the regulation of neuronal activity in these sites. Physiological aging is closely related to the decrease of NOS activity in brain tissue. The decrease of NOS activity leads to a decrease in NO content in the brain, which can lead to a decrease in the phagocytic ability of macrophages; a decrease in the content of active guanine cyclase, a decrease in the ability to regulate various metabolisms, and an accelerated aging. The results of this experiment showed that the NOS activity and NO content of the model aging mice decreased significantly, and LA17 could significantly increase the NOS activity and NO content of aging model mice. The above results suggest that LA17 can increase the production of NO by increasing the activity of NOS, and its biological effects can be sustained, which can delay the aging of the body.

GSH-Px is an important catalytic peroxidation enzyme widely distributed in the body. It specifically catalyzes the reduction of hydrogen peroxide by GSH and inhibits the formation of OH by nicotinamide adenine dinucleotide (NADH). - Impairing the damage to lipid peroxidation, thereby protecting the structure and function of the cell membrane. The results of this experiment show that LA17 can significantly increase the GSH-Px activity in the brain of D-galactose-induced aging mice, increase the activity of antioxidant enzymes in the body, reduce the damage of free radicals to biological macromolecules, and play a role in delaying aging.

In summary, this experiment shows that Lactobacillus acidophilus can increase the levels of SOD, GSH-Px, NO and NOS, and reduce the content of MDA, suggesting that Lactobacillus acidophilus may increase the body's antioxidant capacity and eliminate excessive aging body production. Free radicals inhibit the peroxidation process of the body, tissues and cells, and play a role in delaying aging.

Claims

Lactobacillus acidophilus LA17 having cephalosporin resistance and high expression of Sir2 protein, characterized in that the Lactobacillus acidophilus LA17 is deposited in the Guangdong Provincial Collection of Microorganisms and Cultures under the accession number GDMCC No. 60192 The deposit address is 5th Floor, Building 59, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou, Guangdong, China. The deposit date is May 24, 2017.
Use of the cephalosporin-resistant and high-expressing Sir2 protein of Lactobacillus acidophilus LA17 according to claim 1 for the preparation of an anti-aging food or medicament.
Use of the cephalosporin-resistant and high-expressing Sir2 protein of Lactobacillus acidophilus LA17 according to claim 1 for the preparation of a food or medicament for enhancing immune function.
Use of the cephalosporin-resistant and high-expressing Sir2 protein of Lactobacillus acidophilus LA17 according to claim 1 for the preparation of a food or medicament for regulating intestinal flora.
An anti-aging food comprising an effective amount of the Lactobacillus acidophilus LA17 having a cephalosporin resistance and high expression of a Sir2 protein according to claim 1 and/or a bacterial protein component thereof as an active ingredient.
An anti-aging medicine comprising an effective amount of the cephalosporin-resistant Lartobacillus acidophilus LA17 and/or its bacterial protein component having the cephalosporin resistance and high expression of the Sir2 protein according to claim 1 as an active ingredient, and A pharmaceutically acceptable carrier.
A food for enhancing immune function, comprising an effective amount of the cephalosporin-resistant Lartobacillus acidophilus LA17 having a cephalosporin resistance and high expression of a Sir2 protein as claimed in claim 1, and/or a bacterial protein component thereof .
A pharmaceutical product for enhancing immune function, comprising an effective amount of the cephalosporin-resistant Lartobacillus acidophilus LA17 having a cephalosporin resistance and high expression of Sir2 protein as an active ingredient, and/or a bacterial protein component thereof And a pharmaceutically acceptable carrier.
A food for regulating intestinal flora, comprising an effective amount of the cephalosporin-resistant Lactobacillus acidophilus LA17 having a cephalosporin resistance and high expression of Sir2 protein as claimed in claim 1, and/or a bacterial cell thereof Protein composition.
A medicine for regulating intestinal flora, characterized in that it comprises an effective dose as an active ingredient The Lactobacillus acidophilus LA17 and/or its bacterial protein component of the cephalosporin-resistant and high-expressing Sir2 protein of claim 1, and a pharmaceutically acceptable carrier.