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A kind of preservation method of nitrifying bacteria

A technology of nitrifying bacteria and growth promoters, applied in the direction of preservation of microorganisms, etc., can solve the problems of long cycle, high storage cost, slow growth of nitrifying bacteria, etc., and achieve the effect of slow growth, low storage cost and high survival rate

Active Publication Date: 2020-03-17
CHINA PETROLEUM & CHEM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the slow growth of nitrifying bacteria, it takes a long time to cultivate the bacteria to the stable phase
In addition, during the low-temperature preservation of bacteria, high-concentration nutrients and a large amount of preservatives need to be added, and the preservation cost is relatively high.

Method used

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  • A kind of preservation method of nitrifying bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Prepare a culture solution with an influent ammonia nitrogen concentration of 300mg / L, and use the intermittent activated sludge method to cultivate nitrifying bacteria in a 100L biological aeration reaction tank. The temperature is 27°C, the pH is 7.8, and the dissolved oxygen DO is 2.5-3.5mg / L. Add growth promoter I according to the concentration of the promoter in the culture system 25mg / L. When the cells enter the stable growth phase, the culture is stopped, and the culture time is 3 days. After sedimentation or centrifugation, several washings are performed to collect the cells. Then according to the water content of 50%, mix the bacteria with the prepared preservation nutrient solution, shake well, add 1.5% preservative dimethyl sulfoxide, and store in -50 ℃ ultra-low temperature refrigerator. After 3 years, it was taken out to investigate the activity and survival rate of the bacteria. After 2 days of recovery, the activity of the bacteria can be completely res...

Embodiment 2

[0032] Prepare a culture solution with an influent ammonia nitrogen concentration of 400mg / L, and use the intermittent activated sludge method to cultivate nitrifying bacteria in a 100L biological aeration reaction tank. The temperature is 37°C, the pH is 7.8, and the dissolved oxygen DO is 0.5-1.5 mg / L. Add growth promoter II according to the concentration of the promoter in the culture system is 30mg / L. When the cells enter the stable growth phase, the culture is stopped, and the culture time is 4 days. After sedimentation or centrifugation, multiple washings are performed to collect the cells. Then, according to the water content of 70%, mix the bacteria with the prepared nutrient solution, shake well, add 2.0% preservative dimethyl sulfoxide, and store in a -70°C ultra-low temperature refrigerator. After 5 years, it was taken out to investigate the activity and survival rate of the bacteria. After 4 days of recovery, the activity of the bacteria can be completely restore...

Embodiment 3

[0035] Prepare a culture solution with an influent ammonia nitrogen concentration of 600mg / L, and use the intermittent activated sludge method to cultivate nitrifying bacteria in a 100L biological aeration reaction tank. The temperature is 27°C, the pH is 7.8, and the dissolved oxygen DO is 2.5-3.5 mg / L. Add growth promoter III according to the concentration of the promoter in the culture system 20mg / L. When the bacteria entered the stable growth phase, the culture was stopped, and the culture time was 7 days. After sedimentation or centrifugation, multiple washings were performed to collect the bacteria. Then according to the water content of 60%, mix the bacteria with the prepared nutrient solution, shake well, add 2.5% preservative dimethyl sulfoxide, and store in a -60°C ultra-low temperature refrigerator. After 4 years, it was taken out to investigate the activity and survival rate of the bacteria. After 3 days of recovery, the activity of the bacteria can be completely...

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Abstract

The invention discloses A nitrobacteria preservation method, which comprises the following steps: (1) pouring a growth promoter during the nitrobacteria culture process; culturing to a stationary growth stage, and collecting thalli; (2) preparing a nitrobacteria preservation nutrient solution; (3) mixing the nitrobacteria collected in the step (1) and the preservation nutrient solution prepared in the step (2), and controlling water content to 40-80%; (4) adding a preserving agent; and (5) freezing at low temperature for preservation. The growth promoter contains, by weight, 40-100 parts of metal salt and 5-30 parts of a polyamine substance. The metal salt is composed of calcium salt, copper salt, magnesium salt and / or ferrous salt. By pouring the growth promoter during the nitrobacteria culture process, the thalli can rapidly be proliferated, culture time can be shortened, and especially activity and low temperature tolerance of the thalli can be enhanced. After the low-temperature freezing, the thalli can rapidly regain activity.

Description

technical field [0001] The invention relates to a method for preserving bacteria, in particular to a method for freezing and preserving nitrifying bacteria at low temperature. Background technique [0002] Bacterial strains are one of the main biological resources. After an excellent strain is selected, it must keep its excellent traits unchanged or slow down as little as possible, so as not to reduce the performance of the bacteria and can be used in production for a long time. application. There are many methods of strain preservation. Cryopreservation is an effective way to solve germplasm degradation and prevent natural accumulation mutations, but traditional cryopreservation requires expensive programmed cooling equipment and cumbersome steps. Complete vitrification is a new method of cryopreservation, that is, under high concentration of protective agent, the cells and the protective agent will enter the vitrified state during rapid cooling, avoiding the formation of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04
Inventor 孙丹凤高会杰郭志华赵胜楠李宝忠
Owner CHINA PETROLEUM & CHEM CORP
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