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Medium thermophilic bacillus licheniformis strain and application thereof in treatment on high-temperature oil-bearing wastewater

A technology of Bacillus licheniformis and oily sewage, applied in biological water/sewage treatment, water/sludge/sewage treatment, bacteria, etc., can solve problems such as the application of heat-resistant Bacillus licheniformis

Inactive Publication Date: 2016-06-15
CHANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After reviewing a large number of literature and patent books, it was found that there was no application of Bacillus licheniformis in high temperature (45-60°C) oily wastewater treatment

Method used

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  • Medium thermophilic bacillus licheniformis strain and application thereof in treatment on high-temperature oil-bearing wastewater
  • Medium thermophilic bacillus licheniformis strain and application thereof in treatment on high-temperature oil-bearing wastewater
  • Medium thermophilic bacillus licheniformis strain and application thereof in treatment on high-temperature oil-bearing wastewater

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1: the screening of bacillus licheniformis (Bacillus licheniformis) H43 bacterial strain;

[0025] 1. Sampling: Collect crude oil (outlet pipeline of the three-phase separator, flow for 1 min before sampling) and soil (sampling after removing 0.5 cm of surface soil) from Tainan Oilfield Combined Processing Station in a sterile container, and then store at 4°C for later use .

[0026] 2. Enrichment: Take oil-contaminated soil samples respectively, put them into 250mL triangular Erlenmeyer flasks with 100mL medium (1mL trace element solution plus 10mL mineral salt solution, add sterile water to 100mL) according to the ratio of 5%, shake the table Cultivate for 7d (55°C, 160rpm). Take 5 mL of the enrichment solution and transfer it to the same fresh medium, and culture it under the same conditions for 7 days, so that the enrichment culture is continued for 7 times.

[0027] 3. Separation: Configure LB solid medium, after autoclaving (121°C, 20min), take it out...

Embodiment 2

[0032] Embodiment 2: Morphological and physiological and biochemical characteristics of Bacillus licheniformis (Bacillus licheniformis) H43

[0033] 1. Test method

[0034] Refer to the experimental method of "Bergey's Mannual of Syetematic Bacteriology" to detect its Gram staining, cell size and shape, growth temperature, growth pH range, NaCl tolerance, starch hydrolysis, lipase activity, V-P, methyl red, glucose Acid production experiment.

[0035] 2. Test results

[0036] The results showed that the bacterium was Gram-positive, arranged in a rod shape under the microscope, facultatively aerobic, with a growth temperature of 40-60°C, a growth pH range of 6-9, and a NaCl tolerance of 0%-4%. , V-P is positive, methyl red test is negative, can use glucose to produce acid, hydrolyze starch, use olive oil to emulsify to produce lipase, etc.

Embodiment 3

[0037] Embodiment 3: the 16sDNA identification of bacillus licheniformis (Bacillus licheniformis) H43 bacterial strain:

[0038] 1. Test strain: Bacillus licheniformis (Bacillus licheniformis) H43 of the present invention

[0039] 2. Test method:

[0040] ① Obtain bacteria with good activity: first recover the H43 strain, take 100uL of the frozen storage solution and inject it into 10mL of LB liquid medium, and culture it on a shaker at 55°C and 160rpm for 12-24h. Streak culture the shaken bacterial liquid, culture in a 55°C incubator for 24 hours, pick a single clone on the plate, inoculate it into a test tube of LB liquid medium, shake it at 55°C, and incubate at 160rpm for 12-24h.

[0041] ②Bacterial DNA extraction method (Takara kit)

[0042]③ Bacterial 16SrDNA amplification

[0043] Primers used: Seqforward5'-GAGCGGATAACAATTTCACACAGG-3'

[0044] Seqreverse 5'-CGCCAGGGTTTTTCCCAGTCACGAC-3'

[0045] The results of the identification are attached to the manual figure 2...

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Abstract

The invention discloses a medium thermophilic bacillus licheniformis strain and an application thereof in treatment on high-temperature oil-bearing wastewater. In the invention, a bacillus licheniformis strain is separated from petroleum-contaminated soil from Taizhou oil recovery plant of Sinopec Group Huadong Branch. The bacillus licheniformis strain is 4-5 mm in colony diameter. The colony has a round appearance with tidy edges, has a moist and smooth surface and has a slight-yellow color. The bacteria are in the shape of short bars under an oil microscopy and belong to gram-positive bacteria, have central spores and have no motility. The bacteria are 40-60 DEG C in growth temperature, preferably 50 DEG C, and is positive in both a V-P test and safranine staining and produces high temperature resistant lipase. A fermentation liquid can hydrolyte starch and produce acid. 16s DNA verification proves the bacteria belong to bacillus licheniformis. The invention also discloses the application of the medium thermophilic bacillus licheniformis strain in treatment on the high-temperature oil-bearing wastewater. A test proves the bacillus licheniformis strain can degrade petroleum hydrocarbons at high effect, can reduce energy waste when being used for treating the high-temperature oil-bearing wastewater (45-60 DEG C), and meanwhile reduces environment pollution due to volatilization of aromatic hydrocarbon during a cooling process.

Description

technical field [0001] The invention belongs to the new microbial technology in the field of high-temperature oil extraction sewage treatment, and relates to a medium thermophilic bacillus licheniformis producing high-temperature resistant lipase and its application in high-temperature oily sewage treatment. Background technique [0002] Petroleum is one of the most important energy sources in my country. In 2014, my country's oil output reached 210 million tons, which plays a very important role in production and life. However, a large amount of high-temperature (about 50°C) sewage containing petroleum hydrocarbons is produced during oil exploitation. If it cannot be treated effectively, it will cause great harm to the environment and human health. [0003] The microbial method has the characteristics of low treatment cost, simple operation, good degradation effect of petroleum pollutants, and is not easy to cause secondary pollution, so it has been widely used in the field...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C12N9/20C12R1/10
Inventor 罗明志闫筱王悦邓林红陈皓程旺林姚红生
Owner CHANGZHOU UNIV
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