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Bifidobacterium animalis as well as preparation and application thereof

A technology of animal bifidobacteria and plant lactobacillus, applied in the field of microorganisms, can solve the problems of bacterial cell death and poor acid tolerance

Active Publication Date: 2020-12-18
中科美大(福建)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When placed under low pH conditions, it will lead to a large number of bacterial cell death, especially the bifidobacteria of human origin are very poor in tolerance to acid

Method used

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  • Bifidobacterium animalis as well as preparation and application thereof
  • Bifidobacterium animalis as well as preparation and application thereof
  • Bifidobacterium animalis as well as preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Isolation, Screening and Identification of Animal Bifidobacterium ZK003 Strain

[0028] Feces samples from healthy infants who have not taken drugs recently were collected, fresh feces were preserved with sterilized peptone water, refrigerated at 4°C, and delivered to the laboratory within 4 hours. Use a sterile glass rod to mash the stool sample, dissolve it in sterile water, shake it evenly, take 1ml of the sample solution, add 9ml of sterile diluent, dilute the sample to a suitable gradient according to the ten-fold dilution method, take 1ml of the diluent and add it to Pour the sterilized modified MRS solid medium into the sterilized plate, mix well, cool and solidify, put it into an anaerobic bag, and incubate anaerobically at 37°C for 48h. After multiple screening and cultivation, Bifidobacterium animalis ZK003 was obtained.

[0029] Improved MRS solid medium (per liter): peptone 10.0g, beef extract powder 5.0g, yeast extract powder 4.0g, glucose 20.0g,...

Embodiment 2

[0034] Example 2 Animal bifidobacterium ZK003 tolerance to acid

[0035] Improved MRS liquid medium (per liter): peptone 10.0g, beef extract powder 5.0g, yeast extract powder 4.0g, glucose 20.0g, Tween-801.0ml, dipotassium hydrogen phosphate 2.0g, sodium acetate 5.0g, citric acid Triammonium 2.0g, magnesium sulfate (MgSO 4 7H2O) 0.2g, manganese sulfate (MnSO 4 4H2 O) 0.05g, when the culture medium is sterilized and cooled to 45°C, add a supporting reagent (SR0370) cysteine ​​hydrochloride for every 100ml of MRS medium, and mix well.

[0036] The low-temperature-preserved Bifidobacterium animalis ZK003 strain was activated with the improved MRS solid medium for 2 to 3 generations, then transferred to the sterile improved MRS liquid medium, put into an anaerobic bag, and cultured strictly anaerobically at 37°C for 12 hours Finally, the bacterial solution was inserted into sterile modified MRS liquid medium with different pH values ​​(pH was 2.0, 2.5, and 3.0) with an inoculum ...

Embodiment 3

[0043] Example 3 Bifidobacterium ZK003 tolerance to bile salts

[0044] Animal Bifidobacterium ZK003 strain preserved at low temperature was activated with improved MRS solid medium for 2-3 generations, then transferred to sterile improved MRS liquid medium, put into an anaerobic bag, and cultured strictly anaerobically at 37°C for 12 hours Afterwards, the bacterial solution was inserted into the aseptic modified MRS liquid medium containing different bile salt concentrations (the bile salt concentration was 0.2%, 0.3%, 0.5%, 1.0%) with an inoculum amount of 1%. The modified MRS liquid medium with salt was used as the control, which was also placed in an anaerobic bag, and after strict anaerobic culture at 37°C for 0h, 2h, 4h, 6h, and 24h, samples were taken to determine the number of viable bacteria, and the LOG value of the number of viable bacteria was calculated.

[0045]

[0046] In formula ②: N0 is the number of viable bacteria (CFU / ml) at 0 h; N is the number of viab...

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Abstract

The invention relates to bifidobacterium animalis which is resistant to acid and cholate, can survive at 15-25 DEG C, can regulate intestinal microbial flora and has an inhibiting effect on escherichia coli, salmonella typhimurium and staphylococcus aureus. The bifidobacterium animalis is preserved in CGMCC, has a preservation number of CGMCC No.19827, has the effects of increasing the number of lactic acid bacteria in intestinal tracts and reducing the number of escherichia coli, and can grow in cooperation with lactobacillus rhamnosus CGMCC NO.19826 and lactobacillus plantarum CGMCC NO.19825to jointly regulate the intestinal microbial flora.

Description

technical field [0001] The invention relates to the field of microbes, and more specifically relates to an animal bifidobacterium ZK003 capable of regulating the function of intestinal microbial flora, as well as its preparation and application. Background technique [0002] The human gastrointestinal tract is colonized with various bacteria, which can be divided into beneficial bacteria, harmful bacteria and neutral bacteria according to their impact on human health. Beneficial bacteria are probiotics that have a beneficial effect on the human body. When the probiotics occupy a sufficient number of advantages in the human gastrointestinal tract, they are beneficial to human health. Among them, bifidobacteria are typical beneficial bacteria in the human intestinal tract. A number of research reports at home and abroad have pointed out that bifidobacteria have a variety of physiologically active functions. Their existence can reduce the pH value of the intestinal tract, chang...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A23K10/18A23L33/135A61K35/745A61K35/747A61P31/04C12R1/01C12R1/25C12R1/225
CPCC12N1/20A23K10/18A23L33/135A61K35/745A61K35/747A61P31/04C12R2001/01C12N1/205A61K2300/00Y02A50/30
Inventor 柯媛萍严贤城
Owner 中科美大(福建)生物科技有限公司
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