A kind of petroleum hydrocarbon degrading bacteria and its fermentation method
A technology of petroleum hydrocarbon degrading bacteria and fermentation method, applied in microorganism-based methods, biochemical equipment and methods, bacteria and other directions, can solve the problems of complex bacterial culture process, slow biochemical degradation speed, low degradation efficiency, etc., and achieve easy composition. It has the effect of short fermentation cycle and simple production process.
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Embodiment 1
[0022] A genus of Bacillus brevis (named Brevibacillus parabrevis in Latin literature), strain name: GWN-2, preserved in the General Microorganism Collection Center of China Committee for the Collection of Microorganisms, and the preservation number: CGMCCNo.13903.
[0023] Its gene sequence is shown in the sequence listing.
[0024] Use this type of Bacillus brevus to ferment to obtain petroleum hydrocarbon degrading bacteria products. The specific fermentation method is carried out according to the following steps:
[0025] 1) Streak inoculation of Bacillus pseudobrevius preserved in glycerol at -70°C on a solid medium in a pre-sterilized Petri dish in an ultra-clean workbench. The medium is composed of the following components in terms of mass percentage: 0.4% yeast extract, 0.8% peptone, 1.5% agar powder, and water as the balance, pH 7.0, sterilized at 120°C and 0.11Mpa for 30min. The culture dish was placed upside down in an incubator at 33° C. for 3 days to obtain a sin...
Embodiment 2
[0031] Utilize the class Bacillus brevus of embodiment 1 to carry out fermentation, obtain petroleum hydrocarbon degrading bacterium product, concrete fermentation method is carried out according to following steps:
[0032] 1) Streak-inoculate the Bacillus brevis preserved in glycerol at -70°C on the solid medium in the pre-sterilized petri dish in the ultra-clean workbench (the medium is composed of the following components according to the mass percentage: yeast extract 0.5%, peptone 1%, agar powder 1.8%, the balance is water, pH 7.2, 120℃, 0.11 ± 0.02Mpa continuous sterilization for 30min). The culture dish was placed upside down in an incubator at 33° C. for 3 days to obtain a single colony of activated Bacillus pseudobrevius.
[0033]2) Pick a single colony and suspend it in 20ml sterilized distilled water aseptically. Take 1ml of the sterile pipette and inoculate it on the slant of solid medium in a sterile square bottle with a capacity of 250mL (the medium is composed...
Embodiment 3
[0038] Utilize the class Bacillus brevus of embodiment 1 to carry out fermentation, obtain petroleum hydrocarbon degrading bacterium product, concrete fermentation method is carried out according to following steps:
[0039] 1) Streak-inoculate the Bacillus brevis preserved in glycerol at -70°C on the solid medium in the pre-sterilized petri dish in the ultra-clean workbench. The medium is composed of the following components according to the mass percentage: yeast extract 0.6%, peptone 1.2%, agar powder 2%, the balance is water, pH 7.2. The culture dish was placed upside down in an incubator at 33° C. for 2 days to obtain a single colony of activated Bacillus pseudobrevii.
[0040] 2. Pick a single colony and suspend it in 20ml sterilized distilled water by aseptic operation. Sterilize the pipette to draw 1ml and inoculate it on the inclined surface of the solid medium in a sterile square bottle with a capacity of 250mL, and shake the square bottle gently to spread the singl...
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