Multifunctional petroleum degrading bacterium as well as culture method and application thereof
A technology for oil-degrading bacteria and culture medium, applied in the field of microorganisms, can solve the problems of few reports of multiple functions of Bacillus adenii, and achieve the effects of promoting oil-degrading activity, being beneficial to ecological restoration, and promoting growth.
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Embodiment 1
[0053] Isolation and identification of a strain of Bacillus aryabhattai MG17
[0054] Collect 2g of high-concentration petroleum-contaminated soil in the coastal area of Shengli Oilfield, put it in a 150mL sterile Erlenmeyer flask, add 50mL of sterile inorganic salt medium, and cultivate it at 30°C and 150rpm for 3 days, absorb the bacterial solution, and perform gradient dilution with sterile water , were diluted to 10 -1 , 10 -2 , 10 -3 , 10 -4 , 10 -5 times, each spread 100 μL onto sterile petroleum-inorganic salt solid medium, and culture it statically at 30°C for 3 days, pick a single colony that grows fast and has a large colony to 50mL petroleum-inorganic salt liquid medium, and incubate at 30°C. Cultivate at 150 rpm for 3 days, pipette 100 μL of the bacterial solution and apply it to a sterile petroleum-inorganic salt solid medium, and pick a single colony.
[0055] Take a single colony and send it to a sequencing company for sequencing. After testing, the 16S r...
Embodiment 2
[0084] A culture method for Bacillus aryabhattai (Bacillus aryabhattai) MG17, the steps are as follows:
[0085] The culture medium used in this embodiment:
[0086] Liquid medium, the components per liter include the following: peptone 10g, yeast extract 5g, potassium chloride 3g, magnesium sulfate heptahydrate 3g, sodium chloride 60g, water to 1L, pH natural.
[0087] Solid medium, the components per liter include the following: peptone 10g, yeast extract 5g, potassium chloride 3g, magnesium sulfate heptahydrate 3g, sodium chloride 60g, agar 20g, water to 1L, pH natural.
[0088] (1) Streak Bacillus aryabhattai MG17 on the above-mentioned solid medium, activate it upside down at 32°C for 2 days, and obtain the activated strain;
[0089] (2) Inoculate the activated bacterial strain prepared in step (1) into the above-mentioned liquid culture medium, and culture it on a shaking table for 2 days at 32° C. with a rotation speed of 150 rpm to obtain a seed solution;
[0090] (3...
Embodiment 3
[0093] The application of Bacillus aryabhattai (Bacillus aryabhattai) MG17 in petroleum degradation, the steps are as follows:
[0094] (1) mix the coastal oil-contaminated saline-alkali soil with an oil content of 5% and the petroleum-degrading bacterial agent prepared in Example 2 according to a ratio of 20:1 in mass ratio, and adjust the water content to 20% with sterilized distilled water;
[0095] (2) Keep the water content of the fungus-soil mixture at 20%, and degrade it at room temperature (25±5°C) for 30 days;
[0096] Infrared spectrophotometry (HJ1051-2019) was used to detect the degradation rate of oil in soil. Bacillus aryabhattai (Bacillus aryabhattai) MG17 degraded oil-contaminated soil with an oil content of 5%. After 30 days, the remaining oil content was 3.59%. The rate was 28.2%.
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