A kind of preservation method of nitrous bacteria

A technology of nitrous acid bacteria and ferrous salt, applied in the direction of preservation of microorganisms, etc., can solve the problems of slow growth of nitrous acid bacteria, high storage cost, and long period, and achieve improved stability and antifreeze ability, low storage cost, and solution slow growth effect

Active Publication Date: 2019-07-12
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 nitrous bacteria, it takes a long time to culture the bacteria to the stable growth period
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 nitrous bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Prepare a culture solution with an influent ammonia nitrogen concentration of 400mg / L, and use the intermittent activated sludge method to cultivate nitrous bacteria in a 100L biological aeration reaction tank. The temperature is 32°C, the pH is 7.8, and the dissolved oxygen DO is 1.0-2.5 mg / L. Add growth promoter I according to the concentration of the promoter in the culture system 25mg / L. When the ammonia nitrogen removal rate of the bacteria reaches over 99%, and the nitrification rate reaches 80%, the culture is stopped, and the culture time is 3 days. After sedimentation or centrifugation, several washings are performed to collect the bacteria. 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, the activity and survival rate of the bacteria were taken out to investigate. After 2 day...

Embodiment 2

[0035] Prepare a culture solution with an influent ammonia nitrogen concentration of 600mg / L, and use the intermittent activated sludge method to cultivate nitrous bacteria in a 100L biological aeration reaction tank. The temperature is 35°C, the pH is 8.0, and the dissolved oxygen DO is 1.5-3.0 mg / L. Add growth promoter II according to the concentration of the promoter in the culture system is 30mg / L. When the ammonia nitrogen removal rate of the bacteria reaches over 99%, and the nitrification rate reaches 80%, the culture is stopped, and the culture time is 5 days. After sedimentation or centrifugation, several washings are carried out to collect the bacteria. Then, according to the water content of 60%, mix the bacteria with the prepared nutrient solution, shake well, add 2.0% 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 ...

Embodiment 3

[0038] Prepare a culture solution with an influent ammonia nitrogen concentration of 800mg / L, and use the intermittent activated sludge method to cultivate nitrous bacteria in a 100L biological aeration reaction tank. The temperature is 37°C, the pH is 8.5, and the dissolved oxygen DO is 1.5-2.5 mg / L. Add growth promoter III according to the concentration of the promoter in the culture system 20mg / L. When the ammonia nitrogen removal rate of the bacteria reaches over 99%, and the nitrification rate reaches 80%, the culture is stopped, and the culture time is 7 days. After sedimentation or centrifugation, several washings are performed to collect the bacteria. Then according to the water content of 70%, mix the bacteria with the prepared nutrient solution, shake well, add 2.5% preservative dimethyl sulfoxide, and store in a -70°C ultra-low temperature refrigerator. After 5 years, the activity and survival rate of the bacteria were taken out to investigate. After 4 days of reco...

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Abstract

A preservation method for nitrite bacteria is disclosed. The method includes (1) adding a growth promoter in a nitrite bacterium culture process, culturing until the ammonium-nitrogen removal rate is 99% and the nitrosation rate is 80% and collecting the bacteria, with the promoter including a metal salt, a polyamine compound, an organic acid hydroxylamine and Na<2>SO3 and the metal salt including a calcium salt, a copper salt, a magnesium salt and / or ferrite; (2) preparing a nitrite bacterium preservation nutrition solution; (3) mixing the nitrite bacteria collected in the step (1) with the nitrite bacterium preservation nutrition solution prepared in the step (2), and controlling pH to be 7.5-9.0 and the water content to be 40-80%; (4) adding a preservation agent; and (5) performing low-temperature freezing preservation. Through adding the growth promoter in the nitrite bacterium culture process, the bacteria are rapidly propagated, the nitrosation rate is increased, culture time is shortened, bacterium activity and resistance to low temperatures can be improved particularly, and activity can be restored after low-temperature freezing preservation.

Description

technical field [0001] The invention relates to a method for preserving bacterium, in particular to a method for cryopreserving nitrous bacteria. 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 intracellular ice crystals, an...

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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