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Application method of polyethylene glycol as acidithiobacillus caldus cryoprotectant

A technology of Thiobacillus thermophilus and polyethylene glycol, applied in microorganism-based methods, biochemical equipment and methods, preservation of microorganisms, etc., can solve problems such as protection of protective agents, and achieve the effect of improving survival rate

Inactive Publication Date: 2014-11-05
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, during liquid nitrogen preservation of Thiobacillus thermophile, the protective agent cannot provide effective protection for it, because the survival rate of Thiobacillus thermophile after resuscitation is about 40.1%

Method used

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  • Application method of polyethylene glycol as acidithiobacillus caldus cryoprotectant
  • Application method of polyethylene glycol as acidithiobacillus caldus cryoprotectant
  • Application method of polyethylene glycol as acidithiobacillus caldus cryoprotectant

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Embodiment 1

[0016] Three identified Thiobacillus thermophilic strains (S1, D1, S2) were cultured to mid-logarithmic growth, filtered to remove sulfur powder, collected by centrifugation at 10000r / min for 20min, and washed with dilute sulfuric acid (pH=2.0 ) to wash the bacteria twice, discard the supernatant, and suspend the bacteria sludge with sterile water.

[0017] Preparation of frozen tubes: the obtained bacterial suspension (concentration of 2 × 10 10 per mL) were added to different protective agent solutions at a volume ratio of 1:1, mixed evenly, and then divided into cryopreservation tubes. The concentration of the protective agent is shown in Table 1 below:

[0018] Table 1: Preparation of protective agent concentration

[0019] sterile preservative concentration

20%

40%

60%

80%

100%

Preservative volume

0.5mL

0.5mL

0.5mL

0.5mL

0.5mL

Bacterial suspension volume

0.5mL

0.5mL

0.5mL

0.5mL

0.5mL

...

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Abstract

The invention discloses an application method of polyethylene glycol as an acidithiobacillus caldus cryoprotectant, belonging to the technical field of bacteria preservation. The application method disclosed herein is characterized by mixing a polyethylene glycol solution and an acidithiobacillus caldus bacterium suspension to obtain a mixed solution for cryopreservation, wherein the molecular weight of polyethylene glycol is 2000, 12000 or 20000, and the concentration of polyethylene glycol in the mixed solution is in a range of 10-50%(w / v). According to the invention, when acidithiobacillus caldus is subjected to liquid nitrogen preservation, the preservation effect of polyethylene glycol as the protector is obviously better than that of glycerol, and thus when acidithiobacillus caldus is subjected to liquid nitrogen preservation, polyethylene glycol as the protector can effectively improve a survival rate of acidithiobacillus caldus.

Description

technical field [0001] The invention belongs to the technical field of bacterial preservation, and in particular relates to an application method of polyethylene glycol as a cryoprotectant for Thiobacillus thermophiles. Background technique [0002] Acidithiobacillus caldus is the only moderately thermophilic bacterium in the genus Acidithiobacillus. It is acidophilic and aerobic. 0 ) or metal sulfide ore (S 2- ) as the energy source, with CO in the air 2 It is a carbon source and absorbs inorganic nutrients such as nitrogen and phosphorus to synthesize bacterial cells. It is widely distributed in the acidic water of metal sulfide mines and coal mines. Since it was isolated and identified by HALLBERG and LINDSTROM in 1994, with the continuous understanding of its leaching effect, the application of this bacteria in sulfide ore bioleaching, especially in high temperature bioleaching reactors for metal sulfide concentrates has become more and more more extensive. Researche...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/04C12R1/01
Inventor 吴学玲刘新星刘学端段红胡琪范宏伟张真真刘莉莉
Owner CENT SOUTH UNIV
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