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Acidithiobacillus ferrooxidans freezing-preservation protective agent

A technology of Thiobacillus ferrooxidans and dimethyl sulfoxide, which can be used in the preservation of microorganisms, bacteria, biochemical equipment and methods, etc., can solve problems such as inability to achieve protection effects.

Inactive Publication Date: 2008-05-07
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Acidophilus ferrooxidans is extremely acidophilic and autotrophic, and its special physiological characteristics make it impossible to achieve a good protective effect by using any of the above cryoprotectants alone

Method used

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  • Acidithiobacillus ferrooxidans freezing-preservation protective agent

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Embodiment 1

[0009] 1. Culture and collection of strains

[0010] The acidophilus Thiobacillus ferrooxidans CMS strain was cultivated to the mid-logarithmic growth phase, filtered to remove ferropotassium ferrooxidans, collected by centrifugation at 10,000 r / min for 20 minutes, and washed with dilute sulfuric acid (pH=2.0) twice. After centrifugation at 10000r / min for 5min, discard the supernatant, resuspend the bacteria sludge in sterile water, count with a hemocytometer and dispense into 1.5mL centrifuge tubes (0.5mL each), centrifuge at 10000r / min for 5min, discard the supernatant, After the bacteria sludge was suspended and balanced with 0.5mL protective agent for a period of time, it was quickly frozen in liquid nitrogen for 5 minutes and then transferred to -70°C for storage.

[0011] 2. Determination of Survival Rate

[0012] After being stored at -70°C for 48 hours, it was left at room temperature for 2 hours, counted with a hemocytometer, and the survival rate was calculated as f...

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Abstract

The invention discloses a protector for Acidithiobacillus ferrooxidans in the deep low-temperature freezing reserving course, which comprises the following parts: 100g / L-150g / L glycerine, 80g / L-120g / L trehalose, 160g / L-200g / L saccharose and 40g / L-60g / L dimethyl sulfoxide.

Description

Technical field: [0001] The invention relates to a protective agent in the cryopreservation process of ore leaching microorganism acidophilic Thiobacillus ferrooxidans. Background technique: [0002] Cryopreservation of bacteria is a conventional method of preservation. Although some bacteria have been found to have good antifreeze ability, even without adding any protective agent, the bacteria have a high survival rate after freezing. However, many studies have shown that , Appropriate concentration of protective agent into the cells can avoid freezing damage to the cell membrane, so the survival rate of bacteria added with protective agent is generally greatly improved after freezing. Dimethyl sulfoxide (DMSO) and glycerol are the most commonly used protective agents for cryopreserved strains. For example, the American Type Culture Collection (ATCC) commonly uses a combination of glycerol (10%), DMSO (5%) and culture medium. The prepared mixture preserves most cell lines....

Claims

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Application Information

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IPC IPC(8): C12N1/04C12N1/20C12R1/01
Inventor 邱冠周张燕飞杨宇刘学端柳建设何环师舞阳万民熙
Owner CENT SOUTH UNIV