A kind of oxygen-resistant, acid-resistant, high-sugar-resistant propionibacterium acidogen and its application

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

Active Publication Date: 2019-07-26
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, current legislation sets high standards for food safety, and consumers are more resistant to food with genetically modified microorganisms, so this method of probiotic breeding may be limited in the future

Method used

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  • A kind of oxygen-resistant, acid-resistant, high-sugar-resistant propionibacterium acidogen and its application
  • A kind of oxygen-resistant, acid-resistant, high-sugar-resistant propionibacterium acidogen and its application
  • A kind of oxygen-resistant, acid-resistant, high-sugar-resistant propionibacterium acidogen and its application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] The domestication of embodiment 1 bacterial strain

[0019] (1) Inoculate the original Propionibacterium acidogenicum at a ratio of 5% in 50 mL of seed medium, and culture it at 30 °C and pH 7.0. The original Propionibacterium acidogenicum was purchased from the General Microbiology Center of China Committee for Culture Collection of Microorganisms, with a preservation number of 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 ), 0.05 g; glucose, 5 g; 0.05% resazurin indicator, 2 mL; distilled water to 1000 mL.

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

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

[0024] (2) Inoculate 5% of the bacterial ...

Embodiment 2

[0037] Embodiment 2 strain identification

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

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

[0040] Propionibacterium acidogenicum ( Propionibacterium acidipropionici) L1124 was inoculated in the fermentation medium for culture; by measuring the OD value, the fermentation broth grown to the late logarithmic period was taken, centrifuged at 12,000 rpm for 5 minutes, and the supernatant was removed; 500 μL of Buffer BS was added to resuspend the cells, and 50 μL of Lysozyme (20 mg / mL), fully pipet and mix, incubate in a 37 °C water bath for 1 hour (invert and mix once 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), thoroughly mixed by pipetting, and incubated in a water bath at 56 ℃ for 10 minutes; add 200 ...

Embodiment 3

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

[0046] (1) Analysis of acid resistance;

[0047] Propionibacterium acidogenicum ( Propionibacterium acidipropionici ) L1124 and original Propionibacterium oxytogen were inoculated in 50 mL of fermentation medium at a ratio of 5%, respectively, at 30 °C, with a glucose concentration of 30 g / L, anaerobic fermentation at pH 7.0, pH 6.0, pH 5.0, and pH 4.0. cultivated under 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 ), 0.05 g; glucose, 30 g; distilled water to 1000 mL.

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

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

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Abstract

The invention discloses an oxygen-resistant, acid-resistant, and high-sugar-resistant Propionibacterium acidogen, which is classified as Propionibacterium acidigens ( Propionibacterium acidipropionici ) L1124, deposit number CGMCC No.14869. The bacterial strain of the invention has strong tolerance to oxygen, acid and osmotic pressure; and can overcome the problems of long growth cycle 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P7/52C12R1/01
CPCC12P7/52C12N1/205C12R2001/01
Inventor 黄和李阳江凌朱丽英
Owner NANJING TECH UNIV
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