Oxygen-tolerant, acid-tolerant and high-sugar-tolerant acid-producing propionibacteria and application thereof

A technology of Propionibacterium acidogens and high sugar tolerance, applied in the field of microorganisms, can solve problems such as restriction and rejection, and achieve the effect of strong tolerance and long growth cycle of anaerobic bacteria.

Active Publication Date: 2018-04-20
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, current legislation sets high standards for food safety, and consumers are more resistant to food

Method used

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  • Oxygen-tolerant, acid-tolerant and high-sugar-tolerant acid-producing propionibacteria and application thereof
  • Oxygen-tolerant, acid-tolerant and high-sugar-tolerant acid-producing propionibacteria and application thereof
  • Oxygen-tolerant, acid-tolerant and high-sugar-tolerant acid-producing propionibacteria and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0018] Example 1 Domestication of strains

[0019] (1) Inoculate the original Propionibacterium acid-producing bacteria in 50 mL seed culture medium at a ratio of 5%, and cultivate it at 30 ℃ and pH 7.0. The original Propionibacterium acid-producing was purchased from the General Microbiology Center of the China Microbial Species Collection and Management Committee, with the preservation number CGMCC 1.2232.

[0020] The composition of the seed medium is as follows:

[0021] Yeast extract, 5 g; Tryptone, 5 g; Dipotassium hydrogen phosphate (K 2 HPO 4 ), 0.25 g; Manganese sulfate (MnSO 4 ), 0.05g; glucose, 5 g; 0.05% resazurin indicator, 2 mL; distilled water to make the volume to 1000 mL.

[0022] The required anaerobic environment is formed by high purity nitrogen with a content of 99.999%, which is generally injected to an amount of 1 atmosphere.

[0023] The experiment was repeated three times, the OD of the bacteria 600 The value reaches 2.5.

[0024] (2) Inoculate the bacterial sol...

Example Embodiment

[0037] Example 2 Identification of strains

[0038] Propionibacterium acidogenes ( Propionibacterium acidipropionici ) Sequence determination of L1124 16S rRNA gene

[0039] (1) Extract DNA (refer to TAKARA Genome Extraction Kit)

[0040] Propionibacterium acidigenes ( Propionibacterium acidipropionici ) L1124 was inoculated in the fermentation medium for culture; by measuring the OD value, take the fermentation broth grown to the late logarithmic stage, centrifuge at 12,000 rpm for 5 minutes, and remove the supernatant; add 500 μL of Buffer BS to resuspend the cells, and add 50 μL of Lysozyme (20 mg / mL), mix thoroughly, incubate in a 37°C water bath for 1 hour (invert and mix every 20 minutes); centrifuge at 12,000 rpm for 5 minutes, discard the supernatant; add 180 μL of Buffer GL, 20 μL of Proteinase K (20 mg / mL) and 10 μL of RNase A (10 mg / mL), aspirate and mix thoroughly, incubate in a 56 ℃ water bath for 10 minutes; add 200 μL of Buffer GB and 200 μL 100% ethanol, pipette and...

Example Embodiment

[0045] Example 3 Propionibacterium acidogenes ( Propionibacterium acidipropionici ) Analysis of the growth characteristics of L1124

[0046] (1) Analysis of acid resistance;

[0047] Propionibacterium acidigenes ( Propionibacterium acidipropionici ) L1124 and the original Propionibacterium acidogenic bacteria were respectively inoculated into 50 mL fermentation medium at a ratio of 5%. At 30 ℃, the glucose concentration was 30 g / L, pH 7.0, pH 6.0, pH 5.0, and pH 4.0. Cultivate under oxygen conditions.

[0048] The composition of the fermentation medium is as follows:

[0049] Yeast extract, 10 g; Tryptone, 5 g; Dipotassium hydrogen phosphate (K 2 HPO 4 ), 0.25 g; Manganese sulfate (MnSO 4 ), 0.05g; glucose, 30 g; distilled water to make the volume to 1000 mL.

[0050] The required anaerobic environment is formed by high purity nitrogen with a content of 99.999%, which is generally injected to an amount of 1 atmosphere.

[0051] Three groups of parallel experiments were set up in the exp...

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Abstract

The present invention discloses oxygen-tolerant, acid-tolerant and high-sugar-tolerant acid-producing propionibacteria which is classified and named as Propionibacterium acidipropionici L1124 with thepreservation number of CGMCC. No. 14869. The Propionibacterium acidipropionici L1124 strain has strong tolerance to oxygen, acid and osmotic pressure, and can overcome the problems of long growth period and low yield of anaerobic bacteria in the fermentation process.

Description

technical field [0001] The invention relates to a novel propionibacterium acidogen and its application, and its application in the fermentation industry, which belongs to the field of microorganisms. Background technique [0002] Probiotics are a general term for a class of active beneficial microorganisms that can produce definite health effects, thereby improving the host's micro-ecological balance and exerting beneficial effects. Probiotics are effective in preventing and treating gastrointestinal diseases, helping digestion and absorption, and stimulating the immune system. They are widely used in food, animal husbandry, and medicine. At present, the commonly used probiotic strains mainly include Bifidobacterium, Lactobacillus, Leuconostococcus, Bacillus and Propionibacterium, etc., and each probiotic has its own different "efficacy". At present, the probiotic effect on Propionibacterium has gradually attracted people's attention. The U.S. FDA has announced 42 kinds of...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12P7/52C12R1/01
CPCC12P7/52C12N1/205C12R2001/01
Inventor 黄和李阳江凌朱丽英
Owner NANJING UNIV OF TECH
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