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Method for breeding rapidly growing bifidobacterium species

A bifidobacterium, rapid technology, applied in the field of microbial genetic breeding and fermentation engineering, can solve the problems of limited high-quality probiotics and rare applications of microorganisms, and achieve the effect of genetic stability

Inactive Publication Date: 2018-03-30
SUZHOU JIANSHIXING BIOLOGICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Radiation mutagenesis is a kind of physical mutagenesis. Mutagens mainly include ultraviolet rays, X-rays, γ-rays, and fast neutrons. very limited
There is another type of mutagenesis---chemical mutagenesis. Mutagens are mainly base analogs and alkylating agents, etc., which can randomly generate mutations at the genome-wide level to form SNPs and InDels, which have been relatively well established in plants. Widely used, but few applications in microbiology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Preparation of chemical mutagen solution

[0034] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L nitrosoguanidine stock solution with 0.1M sterile phosphate buffer (pH 7.0).

[0035] (2) Culture medium preparation

[0036] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure 1×10 5 Pa sterilization for 20min.

[0037] MRS plate medium: add 20.0g / L agar on the basis of MRS liquid medium, high pressure 1×105 Pa sterilization for 20min.

[0038] (3) Activation and cultivation of strains

[0039] The Bifidobacterium strain used is Bifidobacterium vulgaris; the strains were streaked and inoculated on the MR...

Embodiment 2

[0045] (1) Preparation of chemical mutagen solution

[0046] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L nitrosoguanidine stock solution with 0.1M sterile phosphate buffer (pH 7.0).

[0047] (2) Culture medium preparation

[0048] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure 1×10 5 Pa sterilization for 20min.

[0049] MRS plate medium: add 20.0g / L agar on the basis of MRS liquid medium, high pressure 1×10 5 Pa sterilization for 20min.

[0050] (3) Activation and cultivation of strains

[0051] The Bifidobacterium strain used is Bifidobacterium vulgaris; the strains were streaked and inoculated on the ...

Embodiment 3

[0057] (1) Preparation of chemical mutagen solution

[0058] First dissolve a certain amount of nitrosoguanidine with a small amount of dimethyl sulfoxide (DMSO), and then prepare a 1.0 g / L nitrosoguanidine stock solution with 0.1M sterile phosphate buffer (pH 7.0).

[0059] (2) Culture medium preparation

[0060] MRS liquid medium: 10g / L peptone, 10g / L beef extract, 5g / L yeast powder, 20g / L glucose, 5g / L anhydrous sodium acetate, 1ml / L Tween-80, 2g / L diamine citrate , 2g / L dipotassium hydrogen phosphate, 0.58g / L magnesium sulfate, 0.25g / L manganese sulfate, dissolved in distilled water, adjusted to pH 6.6-6.8, and constant volume; high pressure 1×10 5 Pa sterilization for 20min.

[0061] MRS plate medium: add 20.0g / L agar on the basis of MRS liquid medium, high pressure 1×10 5 Pa sterilization for 20min.

[0062] (3) Activation and cultivation of strains

[0063] The Bifidobacterium strain used is Bifidobacterium vulgaris; the strains were streaked and inoculated on the ...

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Abstract

The invention discloses a method for breeding rapidly growing bifidobacterium species. According to the method, a chemical mutagen such as nitrosoguanidine is adopted to induce common bifidobacterium,then a relatively scale of bifidobacterium mutation group as a novel bifidobacterium inheritance genetic resource center is developed, a mutation strain which can grow rapidly and is regular in morphology and conspicuous in characteristic is screened from an MRS ((Man Rogosa Sharpe) medium, that is, a bifidobacterium strain which can grow rapidly is bred, the fermentation time is shortened, and the fermentation cost is reduced.

Description

technical field [0001] The invention relates to a breeding method for fast-growing bifidobacterium strains, in particular to the development of mutation populations through chemical mutagenesis to breed new high-quality, fast-growing bifidobacterium strains, and belongs to the technical field of microbial genetic breeding and fermentation engineering . Background technique [0002] There are a variety of flora in the digestive system of the human gastrointestinal tract, including beneficial, harmful and neutral. Under normal circumstances, there is a natural balance of these flora. However, under the modern diet, high pressure of life and work, and pathological conditions, the number of harmful flora increases, and the neutral flora produces harmful substances to the human body, turning into harmful flora, which leads to the natural balance of the human colon and digestive system flora Out of balance, susceptible to infection by pathogenic bacteria, the human body is in a s...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12N1/20C12R1/01
CPCC12N1/20C12N15/01
Inventor 刘世名
Owner SUZHOU JIANSHIXING BIOLOGICAL SCI & TECH CO LTD
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