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Microorganism and bacterial agent thereof, and applications of microorganism and bacterial agent thereof

A microbial agent and bacterial agent technology, applied in the field of microbiology, can solve problems such as poor stability and insufficient biodegradability, achieve small environmental impact, obvious bioremediation effect, and enrich the existing resources of oil pollution degradation strains

Inactive Publication Date: 2018-01-26
BC P INC CHINA NAT PETROLEUM CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to improve the problem of insufficient biodegradability and poor stability in the existing petroleum microbial remediation technology, the object of the present invention is to provide a kind of microorganism Enterobacter hallii Y-29 and the microbial bacterial agent prepared therefrom;

Method used

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  • Microorganism and bacterial agent thereof, and applications of microorganism and bacterial agent thereof
  • Microorganism and bacterial agent thereof, and applications of microorganism and bacterial agent thereof
  • Microorganism and bacterial agent thereof, and applications of microorganism and bacterial agent thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Screening and Identification of Enterobacter hallii Y-29

[0038] 1. Isolation of crude oil bioremediation strains

[0039] Direct separation method:

[0040] (1) Weigh 10g of fresh soil sample and add it into a conical flask filled with 90mL of sterile water, shake at constant temperature for 30min (30°C, 200r / min);

[0041] (2) Add sterilized water for gradient dilution to 10 -6 、10 -7 、10 -8 , take 0.1mL of bacterial solution of three different dilutions and spread them on the oil-containing (0.5%) inorganic salt plate, spread evenly with a sterile spatula, repeat 3 times for each dilution, and culture at 28°C for 4- 5d; Inorganic salt medium formula: NaNO 3 2g, K 2 HPO 4 1g, K 2 HPO 4 ·3H 2 O 0.5g, NaCl 0.5g, MgSO4 7H 2 O 0.1g, CaCl 2 0.01g, FeSO 4 ·7H 2 O 0.01g, add H 2 0 to 1000 mL, pH=7.

[0042] (3) Use an inoculation loop to pick out fast-growing and well-growing colonies and streak them on the LB solid plate, and culture them at a c...

Embodiment 2

[0079] The preparation of embodiment 2 Enterobacter hallii Y-29 inoculum

[0080] The present embodiment provides a kind of preparation method of Enterobacter hallii Y-29 bacterial agent, it comprises the following steps:

[0081] Step 1, inoculating Enterobacter hallii Y-29 in LB solid medium and culturing at 35-37°C for 30-36 hours to obtain an activated strain;

[0082] The preparation method of LB solid medium is: 5g of yeast extract, 10g of tryptone, 10g of sodium chloride, 20g of agar powder, add water to mix and dissolve, adjust the pH value to 7.0-7.2 with sodium hydroxide, and sterilize at 121°C for 20min.

[0083]Step 2: Inoculate the strain activated in Step 1 into LB liquid medium, shake and culture at 37°C and 140r / min for 36h to obtain seed liquid;

[0084] The preparation method of LB liquid medium is: yeast extract 5g, tryptone 10g, sodium chloride 10g, add water to mix and dissolve, adjust the pH value to 7.0-7.2 with sodium hydroxide, and sterilize at 121°C ...

Embodiment 3

[0089] Embodiment 3 Enterobacter hallii Y-29 is to the degradation of oil-contaminated soil

[0090] Weigh 10 g of 3.0%, 5.0%, 7.0%, 9.0%, and 11.0% different oil-contaminated sterilized soils into Erlenmeyer flasks, respectively inoculate 4 mL of bacterial liquid that has been cultured for 24 hours, and incubate at room temperature for 45 days to avoid oil pollution. Contaminated soil was used as control. Dry the soil samples at 105°C and grind them through a 0.25mm sieve. Weigh 5g of dried soil for each treated sample, extract it by Soxhlet extraction method, set the volume to 250mL at a wavelength of 225nm, and use UV spectrophotometry For determination, if the oil concentration of the sample exceeds the range of the standard curve, it is measured after appropriate dilution. 3.0% each treatment takes 1.5mL to 50mL; 5.0% each treatment takes 1mL to 50mL and dilutes 50 times; 7.0%, 9.0% and 11.0% each treatment takes 0.5mL to 50mL and dilutes 100 times. The content of oil i...

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Abstract

The present invention provides a microorganism and a bacterial agent thereof, and applications of the microorganism and the bacterial agent thereof. The microorganism is Enterbacter hormaechei Y-29, wherein the preservation number of the strain is CCTCC NO:M2016049. The preparation method of the Enterbacter hormaechei Y-29 bacterial agent comprises: activating preserved Enterbacter hormaechei Y-29, culturing a seed liquid, fermenting, carrying out centrifuging separation to obtain the Enterbacter hormaechei Y-29, and mixing the Enterbacter hormaechei Y-29 and a Tween solution to obtain the Enterbacter hormaechei Y-29 bacterial agent, wherein the mixing ratio is that per 60 mL of the Tween solution contains 1 g of the bacteria. According to the present invention, the Enterbacter hormaecheiY-29 and the bacterial agent thereof can provide significant bioremediation effects on petroleum pollution, and can provide excellent degradation effects on petroleum in crude oil-polluted and petroleum-polluted soil; and the Enterbacter hormaechei Y-29 bacterial agent has advantages of short production cycle, low cost, simple use, no secondary pollution, low influence on environment and the like,can enrich the existing petroleum pollution degradation strain resources, and has wide application prospects in petroleum-polluted soil remediation.

Description

technical field [0001] The invention belongs to the technical field of microbiology, and relates to a microorganism and its bacterial agent and application. Background technique [0002] With the increasing demand for petroleum products in human society, petroleum and its products enter the environment in various ways, such as oily wastewater from refineries. Foreign refineries produce 0.5-1 ton of oily wastewater when they process 1 ton of crude oil, while Chinese refineries Due to the refining of heavy oil and the complex refining process, 0.7-3.5 tons of oily wastewater is produced for every 1 ton of crude oil processed. According to statistics, my country's oil refining production capacity has reached 2.4×10 per year 8 tons, the actual processing volume is 1.6×10 per year 8 According to this calculation, the annual output of oily wastewater is as high as 1.12-5.6×10 8 Ton. These oily wastewaters cause serious pollution to soil and water quality, threatening human hea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C09K17/14B09C1/10C12R1/01
Inventor 田应兵孙正祥杨佩刘在洲杜显元
Owner BC P INC CHINA NAT PETROLEUM CORP
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