A polycyclic aromatic hydrocarbon degrading microbial agent and its preparation method and application
A technology of microbial agents and polycyclic aromatic hydrocarbons, applied in the field of biological treatment of environmental pollutants, to achieve the effects of a wide range of pH values, super removal capacity, and easy storage and transportation
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Embodiment 1
[0034] The culture method of Mycobacterium flavinum CP13 bacterial strain of the present invention:
[0035] The CP13 strains were streak-inoculated on the nutrient gravy medium, cultured at 28-30°C for 48h, and then respectively inserted into 500mL Erlenmeyer flasks, and cultured at 30°C and 150r / min for 12h. Then it was inserted into a 5L seed tank according to the inoculum amount of 5%, and cultivated for 24h at 180r / min, pH7.5, and ventilation rate 5L / min.
[0036] Nutrient gravy medium, ingredients and dosage (g / L): beef extract 3, peptone 10, NaC15, agar 20, adjust pH 7.0-7.2.
[0037] The cultivation method of Trichoderma viride of the present invention: Inoculate the strain (Trichoderma viride GIM3.141, purchased from Guangdong Microbial Culture Collection Center) into PDA medium, and culture at 30° C. for 5 days.
[0038] PDA medium composition and dosage (g / L): potato 200, glucose 20, agar 20, adjust pH 7.0-7.2.
[0039] Preparation of composite carrier:
[0040] ...
Embodiment 2
[0044] After the bacteria agent prepared in Example 1 was stored at room temperature for a certain period of time, the bacteria agent was inoculated into the MSM culture solution with a pyrene concentration of 10 mg / L in a ratio of 10% (V / V), so as not to contain the bacteria agent The pyrene-containing MSM culture medium was used as a blank, placed in a shaker at 30°C, shaken and cultured at 150r / min in the dark for 2 days, and then the remaining amount of pyrene in the culture medium was determined. Depend on figure 1 It can be seen that the degradation rate of each initial concentration of pyrene maintained at 60%-66% within 170 days of storage at room temperature.
[0045] This example shows that the bacterial agent can be effectively preserved at room temperature, and the effective preservation time is more than 170 days, which provides a guarantee for the bioremediation of the environment polluted by polycyclic aromatic hydrocarbons.
Embodiment 3
[0047] The bacterium agent that embodiment 1 is made is inoculated in the MSM culture fluid of pyrene concentration 10mg / L by the ratio of 10% (V / V), is blank with the pyrene-containing MSM culture fluid that does not contain this bacterium agent, places 30 In a shaker at ℃, under the condition of 150r / min, shaking and culturing in the dark for 2 days, the remaining amount of pyrene in the culture solution was measured. Depend on figure 2 It can be seen that the bacterial agent of the present invention can effectively degrade polycyclic aromatic hydrocarbon pyrene under the condition of pH 4-9, and has a wider pH adaptation range than free bacteria.
[0048] This example shows that the polycyclic aromatic hydrocarbon degrading microbial agent of the present invention can degrade pyrene well in a wide pH range of acidic, neutral and alkaline, which provides a guarantee for its application in different pH environments.
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