Composite microbial agent, and preparation method and application thereof

A technology of compounding microbial inoculants and cells is applied in the field of environmental microbial remediation, which can solve the problems of many polluted sites and difficult remediation.

Inactive Publication Date: 2019-12-10
RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many contaminated sites where Cr(VI) and PAHs are compounded, and the

Method used

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  • Composite microbial agent, and preparation method and application thereof
  • Composite microbial agent, and preparation method and application thereof
  • Composite microbial agent, and preparation method and application thereof

Examples

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Embodiment 1

[0030] The preparation of embodiment 1 composite microbial bacterial agent

[0031] The preparation method of composite microbial bacterial agent in the present embodiment comprises the following steps:

[0032] (1) Preparation of Stenotrophomonas rhizophilus X1 bacterial liquid

[0033] Use an inoculation loop to pick a ring of Stenotrophomonas rhizophila X1 from the thawed tube of glycerol bacteria solution and put it into TSA medium for streak isolation and culture. After 48 hours, when a colony of appropriate size grows on the plate, pick a colony and inoculate it into LB culture medium, and culture it on a shaking table at 150rpm / min at 30°C for 24 hours, as the seed solution, centrifuge the seed solution at 5000r / min for 3min, and collect Bacteria, the collected bacteria were resuspended twice with PBS buffer to adjust the OD of the bacteria solution 600 value to 1.0.

[0034] (2) Preparation of Pseudomonas L1 bacterial liquid

[0035] Use an inoculation loop to pick...

Embodiment 2

[0038] The preparation of embodiment 2 composite microbial bacterial agents

[0039] The preparation method of composite microbial bacterial agent in the present embodiment comprises the following steps:

[0040] (1) Preparation of Stenotrophomonas rhizophilus X1 bacterial liquid

[0041]Use an inoculation loop to pick a ring of Stenotrophomonas rhizophila X1 from the thawed tube of glycerol bacteria solution and put it into TSA medium for streak isolation and culture. After 48 hours, a colony of a suitable size grows on the plate, pick a colony and inoculate it into the LB culture medium, and cultivate it at 180rpm / min at 28°C for 24 hours, as the seed solution, centrifuge the seed solution at 4000r / min for 5min, collect Bacteria, the collected bacteria were resuspended twice with PBS buffer to adjust the OD of the bacteria solution 600 value to 1.2.

[0042] (2) Preparation of Pseudomonas L1 bacterial liquid

[0043] Use an inoculation loop to pick a ring of Pseudomonas ...

Embodiment 3

[0046] The preparation of embodiment 3 composite microbial bacterial agents

[0047] The preparation method of composite microbial bacterial agent in the present embodiment comprises the following steps:

[0048] (1) Preparation of Stenotrophomonas rhizophilus X1 bacterial liquid

[0049] Use an inoculation loop to pick a ring of Stenotrophomonas rhizophila X1 from the thawed tube of glycerol bacteria solution and put it into TSA medium for streak isolation and culture. After 48 hours, when a colony of appropriate size grows on the plate, pick a colony and inoculate it into the LB culture medium, and culture it at 120rpm / min at 32°C for 24h on a shaker, as the seed solution, centrifuge the seed solution at 6000r / min for 2min, collect For the bacterial cells, the collected bacterial cells were resuspended twice with PBS buffer, and the OD600 value of the bacterial liquid was adjusted to 0.8.

[0050] (2) Preparation of Pseudomonas L1 bacterial liquid

[0051] Use an inoculat...

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Abstract

The invention discloses a composite microbial agent, a preparation method and application thereof. The composite microbial agent comprises a stenotrophomonas rhizophila X1 and a pseudomonas L1. The preparation method of the composite microbial agent comprises the steps that (1) the stenotrophomonas rhizophila X1 and the pseudomonas L1 are subjected to pure culture correspondingly, and seed liquidof the two bacteria is correspondingly obtained; (2) the seed liquid of the stenotrophomonas rhizophila X1 and the seed liquid of the pseudomonas L1 are correspondingly centrifuged, and thalli of thetwo bacteria are correspondingly collected; and (3) the thallus of the stenotrophomonas rhizophila X1 and the thallus of the pseudomonas L1 are correspondingly resuspended with a buffer solution, resuspended bacterium liquid is mixed evenly, and the composite microbial agent is obtained. The composite microbial agent is applied to soil in which Cr (VI) and PAHs coexist, and can simultaneously remove the Cr (VI) and the PAHs, the removal rate of the Cr (VI) is as high as 88%, and the removal rate of the PAHs is as high as 55%.

Description

technical field [0001] The invention relates to the technical field of environmental microbial remediation, in particular to a composite microbial bacterial agent and its preparation method and application. Background technique [0002] With the rapid development of the global economy, the problem of heavy metal-organic compound pollution has become more and more prominent, and has become one of the more and more important environmental issues. Chromium salt series products are important basic raw materials for chemical industry-light industry-advanced alloy materials. They are widely used in tanning, electroplating, alloys, pigments, printing and dyeing, offset printing and agriculture. The annual output of chromium salts in my country has exceeded 160,000 tons. The amount of chromium slag discharged is 350,000 to 420,000 tons, and the Cr(VI) contained in it is about 3,500 tons. Cr(VI) can pass through prokaryotic and eukaryotic cell membranes, cause oxidative cell damage, ...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/38C12R1/01
CPCB09C1/10C12N1/20
Inventor 庄绪亮李祥龙吴尚华白志辉
Owner RES CENT FOR ECO ENVIRONMENTAL SCI THE CHINESE ACAD OF SCI
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