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Solid culture medium for separating and purifying extremely thermoacidophilic archaea and preparation method thereof

A solid medium, separation and purification technology, which is applied in the field of solid medium for the separation and purification of extreme acidophilic archaea and its preparation, can solve problems such as the difficulty in realizing the separation and purification of extreme acidophilic archaea, and achieve the effect of broad application prospects

Inactive Publication Date: 2013-06-19
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many disadvantages in the use of these coagulants, and it is also difficult to achieve the separation and purification of extreme acidophilic archaea

Method used

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  • Solid culture medium for separating and purifying extremely thermoacidophilic archaea and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] 1. Synthesis of Medium

[0024] Solution A: 550ml of distilled water, adjust the pH to 2.5, add 2g of jelly powder. The jelly powder was purchased from Richengsheng Food Additive Factory in Changsha, Hunan Province, and the product model was ZG-30B.

[0025] Solution B: 300ml of 9K basal medium, adjust the pH to 1.8, add 5.83g of potassium thiocyanate, and 9ml of yeast powder mother solution (0.2g / L).

[0026] Liquid C: 80 mL of 9K basal medium, add 9.17 g of ferrous sulfate, adjust the pH to 1.8, and filter to sterilize with a 0.22 μm filter membrane.

[0027] Liquid B was sterilized at 121°C for 10 minutes, cooled for 5 minutes, filtered with gauze, mixed with liquid C evenly, and distilled water was added to make up to 1L; plate (9cm) was poured, each plate was about 20ml.

[0028] 2. Bacteria Activation

[0029] The frozen and preserved strains were taken out separately: Acidianus manzaensis and Sulfolobus metallicus (obtained and identified by the biometallurgy ...

Embodiment 2

[0037] Solution A: 450ml of distilled water, adjust pH=3, add 2g of jelly powder. The jelly powder was purchased from Richengsheng Food Additive Factory in Changsha, Hunan Province, and the product model was ZG-30B.

[0038] Solution B: 250ml of 9K basal medium, adjust the pH to 2, add 8g of potassium thiocyanate, 6ml of yeast powder mother solution (0.2g / L).

[0039] Liquid C: 100 mL of 9K basal medium, add 11 g of ferrous sulfate, adjust the pH to 2, and filter to sterilize with a 0.22 μm filter membrane.

[0040] Liquid B was sterilized at 121°C for 10 minutes, cooled for 5 minutes, filtered with gauze, mixed with liquid C evenly, and distilled water was added to make up to 1L; plate (9cm) was poured, each plate was about 20ml.

Embodiment 3

[0042] Solution A: 550ml of distilled water, adjust the pH to 2.5, add 2g of jelly powder. The jelly powder was purchased from Richengsheng Food Additive Factory in Changsha, Hunan Province, and the product model was ZG-30B.

[0043] Solution B: 180ml of 9K basal medium, adjust the pH to 2.5, add 10g of potassium thiocyanate, and 8ml of yeast powder mother solution (0.2g / L).

[0044] Liquid C: 60 mL of 9K basal medium, add 15 g of ferrous sulfate, adjust the pH to 2.5, and use a 0.22 μm filter membrane to filter and sterilize.

[0045] Liquid B was sterilized at 121°C for 10 minutes, cooled for 5 minutes, filtered with gauze, mixed with liquid C evenly, and distilled water was added to make up to 1L; plate (9cm) was poured, each plate was about 20ml.

[0046] The culture medium prepared in the above Examples 2-3 also successfully isolated and cultured single colonies of Acidbacterium manzae and Sulfolobus metallobus.

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Abstract

The invention provides a solid culture medium for separating and purifying extremely thermoacidophilic archaea and a preparation method thereof. The solid culture medium comprises the following components: jelly powder, potassium rhodanide, ferrous sulfate, yeast powder, and a 9K basic culture medium. The preparation method (1L) comprises the following steps of: preparing liquid A, namely regulating the pH value of 400 to 550ml of distilled water to be between 1.8 and 2.5, and adding 2 to 3g of jelly powder; preparing liquid B, namely regulating the pH value of 150 to 300ml of 9K basic culture medium to be between 1.8 and 2.5, and adding 5 to 11g of potassium rhodanide and 5 to 9ml of yeast powder mother liquor (0.2g / L); preparing liquid C, namely adding 9 to 15g of ferrous sulfate into 50 to 100mL of 9K basic culture medium, regulating the pH value to be between 1.8 and 2.5, and filtering and sterilizing by a filter membrane; and sterilizing the liquid A and the liquid B at high temperature respectively, cooling for 5 to 10 minutes, filtering by using a gauze respectively, uniformly mixing with the liquid C, and adding the distilled water until the volume is 1L. The extremely thermoacidophilic archaea can be separated and purified by a plate method on the basis of the culture medium.

Description

technical field [0001] The invention relates to a solid culture medium for separating and purifying extreme acidophilic archaea and a preparation method thereof. Background technique [0002] Extreme acidophilic archaea are one of the hotspots of biogeochemical research at present. They can exist in extreme environments such as high temperature, strong acid / alkaline conditions, high salinity, and hypoxia. plays an important role in. Hyperacidophilic archaea are an independent branch of microbial evolution. Due to their strong iron and sulfur oxidation activity, extreme thermophile and acidophilicity, they are used as model organisms for the study of biometallurgical mechanism, archaeal genetic mechanism and thermophilicity. . Extreme acidophilic archaea mostly exist in sulfur-containing acidic hot springs, which can oxidize ferrous and sulfur. At 60-70°C, they can promote the regeneration of ferric iron as an oxidant, thereby accelerating the dissolution of metals in sulfi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20
Inventor 夏金兰邱冠周巩三强潘佳民聂珍媛梁长利张瑞永
Owner CENT SOUTH UNIV
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