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A strain of Bifidobacterium breve and its cultivation method and use

A technology of Bifidobacterium breve and its cultivation method, which is applied in the field of Bifidobacterium breve and its cultivation, can solve the problems of dairy products and drug efficacy effects, gastric acid resistance and bile salt resistance, etc., and achieve low potential risk, Good resistance to gastric acid and bile salts, and the effect of reducing risks

Active Publication Date: 2022-07-12
AUSNUTRIA DAIRY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, Bifidobacterium breve isolated from existing dairy products and pharmaceuticals is resistant to some antibiotics (such as kanamycin, streptomycin, paromomycin and neomycin, etc.), and resistant to gastric acid And bile salt resistance is not good, which leads to a greater impact on the efficacy of dairy products and medicines containing the above-mentioned Bifidobacterium breve

Method used

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  • A strain of Bifidobacterium breve and its cultivation method and use
  • A strain of Bifidobacterium breve and its cultivation method and use
  • A strain of Bifidobacterium breve and its cultivation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Screening of strains

[0032] 1. Sample collection

[0033] Before sample collection, the nipples of healthy volunteers were subjected to strict sanitation treatment, and then about 10 mL of breast milk was collected and placed in sterile centrifuge tubes, and then stored in a -80°C ultra-low temperature refrigerator.

[0034] 2. Bacteria isolation

[0035] 1) Primary screening of strains

[0036] The breast milk samples after enrichment culture and direct gradient dilution were poured into MRS-Y1 and MRS-Y2 liquid medium, respectively, and cultured in anaerobic bags at 37 °C for 24-48 h until a single colony was grown. The typical morphology of Fidobacteria is a characteristic single colony of white or milky white.

[0037] 2) Strain rescreening

[0038] The strains obtained from the primary screening were streaked in MRS-G1 and MRS-G2 solid medium containing bromocresol violet, and cultured anaerobic for 48 hours. The selected strains with typical bifi...

Embodiment 2

[0064] Example 2: Acid and bile salt tolerance of Bifidobacterium breve grx05

[0065] 1) Test of acid resistance

[0066] Preparation of bacterial suspension: After 24 strains were screened and activated by anaerobic culture, they were respectively inoculated into MRS-Y1 liquid medium (cultivating bifidobacteria) and MRS-Y2 liquid medium ( cultured Lactobacillus), after anaerobic cultivation at 37°C for 24 hours, centrifuged at 6000 × g for 10 min to collect the bacterial cells, and sterilized PBS buffer was added to the bacterial cells to obtain a bacterial suspension, wherein the OD in the bacterial suspension 600 is 1.0.

[0067] Preparation of gastric juice: Dissolve 0.5g NaCl and 0.3g pepsin in deionized water respectively, adjust the pH to 3.0 with 1.0mol / L hydrochloric acid, add deionized water to 100mL, filter and sterilize with a 0.22μm filter, and refrigerate at 4°C spare.

[0068] Take 1.0 mL of bacterial suspension and inoculate it into 9.0 mL of gastric juice ...

Embodiment 3

[0082] Example 3: Inhibitory effect of strain on pathogenic bacteria

[0083] Six pathogenic bacteria, Escherichia coli, Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Salmonella and Pseudomonas were selected as indicator bacteria, and the Oxford cup method was used to determine the inhibitory effect of the strain on the pathogenic bacteria. shown in Table 4.

[0084] Pour thin-layer agar at the bottom of the plate, add the strains isolated in the embodiment of the present invention to the agar; culture the pathogenic bacteria in LB liquid medium at 37°C for 24 hours, and take 0.1 mL of the pathogenic bacteria liquid and spread it on 30 mL of LB. The surface of the solid medium; evenly place a sterilized Oxford cup in a horizontal plate, and draw 0.2 mL of the bacterial suspension to be tested into the Oxford cup. The plate was placed in a refrigerator at 3-4°C for 24h, and then incubated at 37°C for 24h, and the size of the inhibition zone was measured. Repeat th...

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Abstract

The present invention provides a strain of Bifidobacterium breve and a method for culturing and use thereof. The Bifidobacterium breve is classified and named Bifidobacterium breve grx05, which has been preserved in China Common Microbial Bacteria on January 4, 2021 Species Preservation and Management Center, the preservation number is CGMCC No. 21589, and the preservation address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, China. The Bifidobacterium breve provided by the invention is not only sensitive to most antibiotics, but also has better resistance to gastric acid and bile salt, thereby reducing its potential risks and making it have a relatively broad application prospect.

Description

technical field [0001] The present invention relates to the technical field of microorganisms, in particular, the present invention relates to a strain of Bifidobacterium breve and a culture method and use thereof. Background technique [0002] Probiotics are a class of active microorganisms that are beneficial to the host. They are colonized in the human intestine and reproductive system, and can produce exact health effects to improve the host's microecological balance and play a beneficial role in the intestine. The general term for active beneficial microorganisms . However, with the widespread use and even abuse of antibiotics, probiotics have developed resistance to some antibiotics and their resistance genes, which have the potential to be transferred to other intestinal flora and pathogenic bacteria. Other intestinal flora and pathogenic bacteria with the above-mentioned resistance genes may not be affected by antibiotics, thus seriously endangering the health of th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23C9/123A23L33/135A23L2/38A23L7/104A23C9/152C12R1/01
CPCC12N1/20A23C9/1234A23L33/135A23L2/382A23L7/104A23C9/152A23V2002/00A23V2400/519A23V2200/30Y02A50/30
Inventor 汪家琦顾瑞霞潘丽娜张臣臣康文丽胡鹏钰吴忠坤颜卫彬戴智勇彭灿
Owner AUSNUTRIA DAIRY CHINA
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