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Method for culturing sporosarcina pasteurii with high urease activity

A technology of Pasteurella and Sarcina, applied in the field of microorganisms, can solve the problems of easy injury, high randomness, and danger to operators, and achieve the improvement of the urease activity of Pasteurella Sarcina, high repetition rate, The result is a stable effect

Inactive Publication Date: 2018-02-23
NORTH CHINA UNIV OF WATER RESOURCES & ELECTRIC POWER
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the mutagenesis method is complicated to operate and has high randomness. Under alkaline conditions, diazomethane will be formed during the NTG mutagenesis process, which will cause an explosion after being hit, and the operation process is extremely dangerous; and nitrosoguanidine is a A strong carcinogen, which can easily cause harm to operators

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  • Method for culturing sporosarcina pasteurii with high urease activity

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Embodiment 1

[0020] A method for culturing Pasteurella Pasteurella with high urease activity, the concrete steps are as follows:

[0021] (1) Prepare A liquid medium and B liquid medium:

[0022] A liquid medium includes sodium citrate 2-5 g / L, peptone 2-5 g / L, beef extract 2-5 g / L, urea 15-20 g / L, nickel chloride 0.02-0.05 g / L, pH value is 7-9;

[0023] B liquid medium includes yeast extract 20-30 g / L, ammonium chloride 10-15 g / L, nickel chloride 0.02-0.05 g / L, pH 7-9;

[0024] (2) After adding agar powder to the liquid medium A and mixing evenly, pour the plate until it solidifies into a solid state, streak the solid plate of medium A with the mother liquor of Pasteurella sporogenes, and cultivate at a constant temperature for 12-16 hours; Then single colonies are picked from the streak solid plate, placed in a test tube containing A liquid medium, and cultured under aerobic conditions for 12-16 hours at a constant temperature of 27°C-37°C;

[0025] (3) Take the bacterial liquid after...

Embodiment 2

[0030] A method for culturing Pasteurella Pasteurella with high urease activity, the concrete steps are as follows:

[0031] (1) Prepare A liquid medium and B liquid medium:

[0032] A liquid medium includes sodium citrate 2 g / L, peptone 2 g / L, beef extract 3 g / L, urea 15 g / L, nickel chloride 0.02 g / L, pH 8;

[0033] B liquid medium includes yeast extract 20 g / L, ammonium chloride 15 g / L, nickel chloride 0.02 g / L, pH 8;

[0034] (2) Add 1% agar powder to the liquid medium A and mix well, pour the plate until it solidifies into a solid state, streak the solid plate of medium A with the mother liquor of Pasteurella sporogenes, and cultivate at a constant temperature of 32 °C for 12 hours; then single colonies were picked from the streaked solid plate, placed in a test tube containing 5 ml of A liquid medium, and cultured in a shaking incubator for 12 hours at a constant temperature of 32 °C and a rotational speed of 180 r / min;

[0035] (3) Take the bacterial liquid after shaki...

Embodiment 3

[0040] A method for culturing Pasteurella Pasteurella with high urease activity, the concrete steps are as follows:

[0041] (1) Prepare A liquid medium and B liquid medium:

[0042] A liquid medium includes sodium citrate 5g / L, peptone 5g / L, beef extract 5 g / L, urea 20g / L, nickel chloride 0.05g / L, pH 8; B liquid medium includes yeast extract 30g / L, ammonium chloride 15 g / L, nickel chloride 0.05 g / L, pH value 8;

[0043] (2) Add 1% agar powder to the A liquid medium and mix it evenly, pour the plate until it solidifies into a solid state, streak the A medium solid plate with the mother liquor of Pasteurella sporogenes, and incubate at 37°C for 15 minutes. Then, single colonies were picked from the streaked solid plate, placed in a test tube containing 5 ml of A liquid medium, and cultured in a shaking incubator for 15 hours at a constant temperature of 37 °C and a rotation speed of 180 r / min;

[0044] (3) Take the bacterial liquid after shaking culture in step (2), inoculate...

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Abstract

The invention discloses a method for culturing sporosarcina pasteurii with the high urease activity. The method particularly comprises the following steps that a liquid culture medium A and a liquid culture medium B are prepared; agar powder is added into the liquid culture medium, mixing is conducted to be uniform, and a sporosarcina pasteurii mother solution is used for conducting streak cultureon a culture medium A solid plate; a single colony is picked for culture; a bacterium solution obtained after shaking culture is taken and inoculated into a conical flask loaded with the liquid culture medium B, enlarged culture is conducted, and a sporosarcina pasteurii solution high in urease activity can be obtained. Accordingly, double culture media of the sequence of A and B are adopted forculturing the sporosarcina pasteurii, the culture medium A is adopted for the single colony culture and strain reactivation stage, and the culture medium B is adopted for the stain enlarged culture stage. The results prove that by means of the method, the sporosarcina pasteurii urease activity is greatly improved, operation is easy, the results are stable, and the repetition rate is high.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a method for culturing Pasteurella Pasteurella with high urease activity. Background technique [0002] The biochemical process of urea catalyzed by urease-producing microorganisms is the core of microbial-induced carbonate precipitation technology. This process can be used to induce or control a series of chemical reactions in rock and soil, thereby improving the properties of geotechnical engineering. It belongs to biology and civil engineering. of cross-research applications. Sporosarcina pasteurii, also known as pasteurii or pasteurii, is one of the most commonly used microorganisms in this technology, but traditional general culture methods such as NH4-YE medium are used. When cultured in a single medium, the yield of urease by S. pasteurii strains was low, and the results obtained by the conductivity method were generally only 8-15 U / ml. [0003] In ord...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
CPCC12N1/20
Inventor 赵阳程留全吕杰刘娉慧黄志全张婧范存彬
Owner NORTH CHINA UNIV OF WATER RESOURCES & ELECTRIC POWER
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