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A medium for screening nitrogen sources suitable for the proliferation of bifidobacteria

A technology of bifidobacteria, proliferation and culture, applied in the field of fermentation engineering, can solve the problems of rapid, efficient and accurate screening of unfavorable nitrogen sources, time-consuming and labor-intensive, and poor osmotic pressure resistance of bifidobacteria.

Active Publication Date: 2020-09-04
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But this method has introduced the use of fermentor, expends a large amount of time and manpower, in addition because bifidobacteria are poor in osmotic pressure resistance, the osmotic pressure is too large when growing to the highest point in the constant pH sugar supplement culture of fermenter, will greatly Inhibit the growth of strains, therefore, constant pH sugar supplementation culture in fermenters is not conducive to the rapid, efficient and accurate screening of nitrogen sources

Method used

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  • A medium for screening nitrogen sources suitable for the proliferation of bifidobacteria
  • A medium for screening nitrogen sources suitable for the proliferation of bifidobacteria
  • A medium for screening nitrogen sources suitable for the proliferation of bifidobacteria

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preparation example Construction

[0041] (7) Preparation of seed liquid: inoculate the culture liquid of bifidobacterium (Bifidobacterium adolescent Z25 or Bifidobacterium bifidum F35) activated once into MRS liquid medium with an inoculum size of 1-5%, and inoculate at a constant temperature of 37 ℃ anaerobic incubator for activation and cultivation for 18-24 hours. Then centrifuge to remove the supernatant, wash once with sterile physiological saline, add the same amount of normal saline as the culture medium to resuspend the bacteria, and use it as the seed liquid for nitrogen source screening and cultivation.

[0042] (8) MRS medium composition: yeast powder 5g / L, anhydrous sodium acetate 5g / L, beef extract 10g / L, anhydrous glucose 20-30g / L, peptone 10g / L, magnesium sulfate heptahydrate 0.25g / L , dipotassium hydrogen phosphate 2g / L, manganese sulfate monohydrate 0.1g / L, diammonium hydrogen citrate 2g / L, Tween-80 1mL / L, adjust the pH to 6.0, and additionally add cysteine ​​hydrochloride 1.0g / L.

[0043] T...

Embodiment 1

[0044] The nitrogen source screening of embodiment 1 Bifidobacterium adolescentis Z25

[0045] (1) Traditional methods for screening nitrogen sources

[0046] Medium preparation: 25-40g / L different nitrogen sources (yeast extract, yeast extract powder 803, yeast extract powder 528, yeast protein 103, soybean peptone, fish bone peptone, tryptone, fish peptone, beef peptone, bovine bone peptone peptone, beef extract powder, beef extract), replace all nitrogen sources (tryptone, yeast powder and beef extract) in the MRS medium respectively, and prepare medium with different nitrogen sources. Heat to dissolve, and sterilize at 115°C for 20 minutes.

[0047] Inoculate Bifidobacterium adolescent Z25 with 2% inoculum amount, place it in an anaerobic incubator at 37°C for static culture, and take samples to measure OD after the strain grows to the stable stage 600 , when the absorbance value exceeds 0.8, dilute the bacterial suspension by a certain factor, so that the diluted absorb...

Embodiment 2

[0073] Embodiment 2: Nitrogen source screening of Bifidobacterium bifidum F35

[0074] The method is the same as that in Example 1, except that the strain is changed to Bifidobacterium bifidum F35.

[0075] (1) Traditional methods for screening nitrogen sources

[0076] Table 4 The results of Bifidobacterium bifidum F35 traditional nitrogen source screening

[0077]

[0078]

[0079] The results showed that the glucose content of the fermented liquid was all≦1.0g / L at the end of the cultivation, indicating that the glucose in each nitrogen source medium was almost completely utilized, and the pH value of the fermented liquid had dropped to 3.5-4.2 (bifidobacteria with higher acid resistance Poor, growth inhibited below pH 4.2) (Table 1); indicating that when the bacteria grow to the stationary phase, it is because of carbon source deficiency or acid inhibition. The nitrogen source may not be fully utilized, and how much nitrogen source the bacteria have used to prolife...

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Abstract

The invention discloses a culture medium for screening nitrogen sources suitable for bifidobacterium proliferation, and belongs to the technical field of fermentation engineering. Culture mediums of different nitrogen sources can be prepared and obtained by using the culture medium formula of the invention, in the culture mediums of different nitrogen sources, the pH value of the fermentation liquid is not less than 4.5 when the bacterial concentration of bifidobacterium adolescentis Z25 and bifidobacterium bifidum F35 is the highest, and the glucose content is not less than 2.0g / L. Moreover,the osmotic pressure of the culture medium is 300-400 mOsm / kg, indicating that the growth of bacteria is stopped because the effective nitrogen source is completely utilized, not because of lack of carbon source or acid or osmotic pressure inhibition. Therefore, the growth rate of bacterial strains using different nitrogen sources and the increment of strains using different nitrogen sources per unit mass can be clearly judged. The problems of traditional batch culture acid inhibition, substrate deficiency inhibition or osmotic pressure inhibition, and osmotic pressure inhibition of nitrogen source screened by sugar supplementation culture at constant pH in fermentation tank can not accurately analyze the screening of effective nitrogen sources are solved.

Description

technical field [0001] The invention relates to a culture medium for screening nitrogen sources suitable for the proliferation of bifidobacteria, belonging to the technical field of fermentation engineering. Background technique [0002] Bifidobacteria widely exist in the intestines of animals and humans, and are very important genus of intestinal microorganisms. In recent years, the probiotic properties of Bifidobacteria have received a lot of attention and research at home and abroad, and have become very important probiotics. However, there are few types of Bifidobacterium strains successfully commercialized at present, and the product form is less diverse than Lactobacillus commercial products, and the content of live bacteria is also low. The price of bifidobacterium products is also very expensive, and the price of its freeze-dried powder is several times that of lactobacillus. This is because the preparation of bifidobacteria is very difficult, and the osmotic pressu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/04
Inventor 陈卫崔树茂朱丹凤毛丙永陆文伟翟齐啸赵建新张灏
Owner JIANGNAN UNIV
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