A kind of multifunctional petroleum degrading bacteria and its culture method and application
A technology of petroleum-degrading bacteria and its cultivation method, which is applied in the field of petroleum-degrading bacteria and its cultivation, can solve the problems of few reports on multiple functions of Bacillus arborii, and achieve the goal of promoting petroleum-degrading activity, promoting growth, and facilitating ecological restoration Effect
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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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