In-situ environment microbe isolation method and soil source petroleum degrading microbe isolation and screening method

A technology of environmental microorganisms and separation methods, which is applied in the field of in-situ environmental microorganism separation, soil oil-degrading microorganism separation and screening, and can solve the problems of time-consuming, labor-intensive, different growth speeds of microorganisms, and environments that cannot provide material exchange.

Inactive Publication Date: 2017-09-01
李宜芳
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are many defects in the traditional method of microorganism isolation and cultivation: first, it cannot meet the growth conditions of microorganisms in the natural environment; second, in the natural environment, microorganisms do not exist in isolation, but co-exist with other microorganisms, through direct contact or material exchange. Form a mutual relationship, and the traditional pure culture cannot provide an environment for material exchange; third, the growth rates of differe

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  • In-situ environment microbe isolation method and soil source petroleum degrading microbe isolation and screening method
  • In-situ environment microbe isolation method and soil source petroleum degrading microbe isolation and screening method
  • In-situ environment microbe isolation method and soil source petroleum degrading microbe isolation and screening method

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Embodiment 1

[0031] This embodiment mainly introduces the structural composition and working principle of an in-situ environmental microorganism cultivation device. Such as Figure 1 to 3 As shown, the present invention provides an in-situ environmental microorganism cultivation device, which includes an upper cover 1, a lower cover 3 and a splint 2. Preferably, the horizontal projection shapes of the upper cover plate 1, the lower cover plate 3, the splint 2 and the microporous filter membrane 4 are all squares of the same size, and the upper cover plate 1 and the lower cover plate 3 have a flat surface. The thickness of the edge on the other side is greater than the thickness at the position where the through hole is provided in the middle, which can effectively increase the strength of the edge, thereby improving the overall strength and reliability of the device.

[0032] The upper cover 1, the lower cover 3, and the splint 2 are provided with a plurality of longitudinal through holes 5 ...

Embodiment 2

[0038] This embodiment mainly introduces a method for separating microorganisms from petroleum produced liquid. For the petroleum produced fluid collected from Jilin Oilfield, the cultivation device described in Example 1 was used to separate functional microorganisms in situ. Proceed as follows:

[0039] 1. Prepare a solid medium containing 2% agar by taking an appropriate amount of extraction liquid;

[0040] 2. Take part of the petroleum produced fluid and record the microbial concentration with a hemocytometer (the actual measured concentration is about 7.6*10 7 Pcs / mL);

[0041] 3. Gradient dilution. Take part of the diluted solution and mix it into the 2% solid medium prepared in step 1 (cooled to about 45°C) to make the final concentration of bacteria 10 3 Pieces / mL (ie 1 piece / μL, so that there is only one cell in each well on average);

[0042] 4. Put the splint into the above-mentioned mixed culture medium and cool it until it solidifies; quickly take out the splint and le...

Embodiment 3

[0052] This embodiment mainly introduces a method for separating soil microorganisms using the culture device described in Embodiment 1. For soil samples collected near Weiming Lake of Peking University, a culture device was used to separate microorganisms in the soil samples in situ. Proceed as follows:

[0053] 1. Put 1g of soil into 10mL sterile water and shake for 10min to obtain soil extract;

[0054] 2. Take part of the soil extract and record the microbial concentration with a hemocytometer (the actual measured concentration is about 4.8*10 7 Pcs / mL);

[0055] 3. Gradient dilution. Take part of the dilution and mix it into 2% LB solid medium (cooled to about 45°C) to make the final concentration of bacteria 10 3 Pieces / mL (ie 1 piece / μL, so that there is only one cell in each well on average);

[0056] 4. Put the splint into the above-mentioned mixed bacteria culture medium and cool it until it solidifies; quickly take out the culture device and leave the small pieces of agar...

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Abstract

The invention discloses an in-situ environment microbe isolation method and a soil source petroleum degrading microbe isolation and screening method. By culturing bacteria in a micropore culturing room, isolation and constraint to different types of microbial cells are realized, and moreover free exchange of water, soluble substances and substances produced during microbial metabolism and decomposition in an in-situ environment is further realized. The methods take petroleum degrading function microbial isolation and screening as a target, so as to perform culturing and enrichment and high throughput screening on target microbes, and compared with the traditional method, the methods have the advantage that more bacterial strain types can be found and gathered.

Description

Technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a microorganism cultivation method, in particular to an in-situ environmental microorganism separation method and a soil-origin petroleum degradation microorganism separation and screening method. Background technique [0002] A large number of microorganisms exist in the natural environment. The separation and cultivation of microorganisms is an important basis for the research and utilization of microorganisms. However, the traditional methods of separation and cultivation of microorganisms have many shortcomings: one is that they cannot meet the growth conditions of microorganisms in the natural environment; the other is that in the natural environment, microorganisms do not exist in isolation, but co-exist with other microorganisms through direct contact or material exchange. Forms a mutual relationship, and traditional pure culture cannot provide an environment for material ...

Claims

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Application Information

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IPC IPC(8): C12N1/02C12N1/20C12M1/00B09C1/10
CPCB09C1/10C12M25/04C12N1/02C12N1/20
Inventor 李宜芳
Owner 李宜芳
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