A strain of thermophilic facultative anaerobic microorganism and its application
A facultative anaerobic and microbial agent technology, applied in the field of microbial enhanced oil recovery, can solve the problems of lowering the viscosity and freezing point of crude oil, poor effect, and inability to degrade high pouring oil, so as to enhance fluidity and improve recovery rate effect
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Embodiment 1
[0036] Embodiment 1, the screening of thermophilic facultative anaerobic strain 64-1
[0037] Take 5 mL of oil well output fluid from a block in Liaohe Oilfield and inoculate it into a 200 mL Erlenmeyer flask containing 100 mL of enrichment medium, wherein the components of the enrichment medium include: corn steep liquor dry powder 0.4%, diammonium hydrogen phosphate 0.2 %, sodium nitrate 0.2%, yeast powder 0.05%, 150rpm, 70 ℃ constant temperature shaker culture for 7 days, take 10ml culture solution and put it into a 200mL Erlenmeyer flask with a capacity of 100mL enrichment medium for cultivation, repeat 5 Second-rate.
[0038]The obtained enrichment was serially diluted, and 0.1 mL was spread on a solid enrichment medium (containing 1% agar) plate medium, and cultured at 70°C. Then, typical colonies were picked and streaked 3 to 4 times repeatedly to obtain colonies of a single form.
Embodiment 2
[0039] Example 2, identification of thermophilic facultative anaerobic strain 64-1
[0040] A single colony on the agar solid medium in Example 1 was picked with a toothpick, and the 16S rDNA gene of the strain was amplified, using universal primers 27F and 1492R, as shown in Table 1. The reaction system is 50 μL: 2×Taq PCR mix 25 μL, 27F 1.0 μL, 1492R 1.0 μL, ddH 2 O 23.0 μL. The PCR reaction program was: denaturation at 94°C for 5min, 94°C for 30S, 57°C for 30S, 72°C for 1min and 30S, 30 cycles, extension at 72°C for 10min, and storage at 4°C.
[0041] Use 0.8% agarose gel electrophoresis to detect the PCR amplification product, sample: 3 μL PCR product + 1 μL Gel-Red, the electrophoresis condition is 150V 15min. Samples with positive test results were sent to BGI for sequencing.
[0042] Table 1
[0043]
[0044] The full-length sequence of the 16S rDNA gene of bacterial strain 64-1 is 1453bps. According to the comparison results of GenBank, the similarity between th...
Embodiment 3
[0045] Example 3, Morphological characteristics, physiological and biochemical experiments of thermophilic facultative anaerobic strain 64-1
[0046] 1. Test strains
[0047] The thermophilic facultative anaerobic strain 64-1CGMCC 1.16798 isolated in Example 1 of the present invention.
[0048] 2. Experimental method
[0049] Referring to the experimental method of "Bergey's Manual of Systematic Bacteriology", the Gram staining, cell size, shape, growth temperature, pH and sodium chloride tolerance range of the tested strains were detected, and the anaerobic characteristics of the strains were detected by the puncture test.
[0050] 3. Experimental results
[0051] Morphological characteristics of the tested strains: Gram staining was negative. Such as figure 2 As shown, the cells under the scanning electron microscope are short rod-shaped, without flagella, and the cell size is 0.6-0.8 μm×1.0-1.2 μm.
[0052] Such as image 3 As shown, the physiological and biochemical...
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