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Process for storing methane oxidizing bacteria

A technology of methane-oxidizing bacteria and bacteria, applied in the direction of using microorganisms, etc., can solve the problems of time-consuming, labor-intensive, inconvenient implementation, etc., and achieve the effect of reducing the chance of pollution

Inactive Publication Date: 2006-10-11
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method can avoid bacterial contamination and maintain a high activity of methane oxidation, it is time-consuming and laborious, and it is very inconvenient to implement

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1, the liquid culture of methanotrophic bacteria

[0017] 1. Obtaining a single colony of methanotrophs

[0018] Use a sterilized inoculation loop to pick up the methanotrophic bacteria preservation solution (Methylosinus trichosporium) OB3b (Sullivan JP et al. Methanotrophs, Methylosinus trichosporium OB3b, sMMO, and their application to bioremediation, Crit.Rev.Microbiol., 1998, 4 (24 ): 335-373; Takeguchi M et al., Properties of the membranes containing the particulate methane monooxygenase from Methylosinus trichosporium OB3b, Biometals, 1998, 11: 229-234), inoculated it on the NMS plate solid medium by streaking On the surface, 5-10 ml of methane gas was added to the culture dish through a 0.2 μ filter every day with a syringe, and after four weeks of cultivation at 30 ° C, obvious colonies were produced on the plate.

[0019] 2. Liquid culture of methanotrophs

[0020] 1) Take a 50 ml culture bottle, fill it with 10 ml of NMS liquid medium, and steril...

Embodiment 2

[0022] Embodiment 2, the preservation of methanotrophic bacteria in paraffin oil

[0023] Pour 1.5 milliliters of the methane-oxidizing bacteria cultured in Example 1 into a sterilized 1.5 milliliter centrifuge tube, centrifuge at 8000 rpm for 10 minutes, discard the supernatant, and collect the The cells were added to 500 microliters of paraffin oil sterilized (heated at 121°C for 20 minutes), oscillated on a vortex shaker to disperse the cells evenly in the paraffin oil, and then stored in a -20°C refrigerator.

Embodiment 3

[0024] Embodiment 3, the use of the methanotrophic bacteria preserved in paraffin oil

[0025] Take a 50ml culture bottle, fill it with 10ml of NMS liquid medium, and after sterilizing it at 121°C for 20 minutes, take a tube of methane oxidizing bacteria preserved in paraffin oil for 90 days from the refrigerator, and put the thawed bacterial solution in the ultra-clean bench Pour it all into the NMS liquid medium, add 5-20ml of methane gas through a 0.2μ filter every day with a syringe, shake and culture at 30°C and 150rpm for three days, the medium appears cloudy, and its OD 600 If the value is about 0.1-0.3, the methane-oxidizing bacteria are successfully cultivated, which shows that the preservation of the methane-oxidizing bacteria by the method of the present invention can maintain a high biological activity of the bacteria for a long time.

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PUM

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Abstract

This invention discloses a method for preserving methane-oxidizing bacteria. The method comprises: collecting the thallus of methane-oxidizing bacteria, mixing with sterilized liquid petrolatum, and stored at -4deg C--90deg C. Since the liquid petrolatum can not be used as carbon sources for most hybrid bacteria, this method can dramatically reduce the chance of contamination. Furthmore, it can be easily operated by directly adding the mixture of bacteria and liquid petrolatum to the culture media. This invention will play important roles in methane-oxidizing bacteria preservation and the industrial procuction thereof.

Description

technical field [0001] The invention relates to a method for preserving microorganisms, in particular to a method for preserving methane-oxidizing bacteria that can effectively prevent contamination by miscellaneous bacteria. Background technique [0002] Methanotrophs are a very special kind of microorganisms in nature, which use methane as the only carbon source and energy source. All methanotrophs are incapable of utilizing multi-carbon compounds, although some methanotrophs can also utilize methanol simultaneously. Studies have shown that methanotrophs can be widely used in the large-scale industrial production of chemical products (natural gas or biogas, etc.) that use methane as raw material, and can also be used in the treatment of organic pollutants in the environment, and can even be used to effectively prevent coal mines from Gas explosion in gas. At present, there is no special preservation method for this strain, and it is generally preserved at -70°C after add...

Claims

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Application Information

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IPC IPC(8): C12N1/26
Inventor 缑仲轩邢新会赵洪新吴昊韩冰江皓王磊
Owner TSINGHUA UNIV