Screening and application of benzo[a] anthracene degrading bacterium in black and odorous bottom mud
A technology of black and smelly bottom mud and benzo, which is applied in the field of microbial degradation technology and environmental protection, and can solve the problems of few researches on benzo[a]anthracene degrading bacteria.
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Embodiment 1
[0033] [Example 1] Domestication and screening of benzo[a]anthracene degrading bacteria
[0034] 1. Acclimatization and cultivation of bacterial strains
[0035] The samples in this experiment were collected from the black and odorous bottom mud of a section of the Maozhou River in Shenzhen City, Guangdong Province. Immediately after collection, they were stored in a 4°C cold storage in the laboratory away from light. Dissolve 5 g of bottom mud in 10 mL of sterile water, shake at a constant temperature for 24 hours, and inoculate 1 mL of bottom mud leachate into 50 mL of liquid medium with a concentration of 1 mg / L of benz[a]anthracene. Under the condition of about 30°C, cultivate them on a shaker at a speed of 140r / min for 7 days. Get 2mL bottom mud enrichment liquid in 50mL benz[a] anthracene concentration and be the liquid culture medium of 10mg / L (medium composition: 2.0g / L (NH 4 ) 2 SO 4 , 0.2g / L MgSO 4 ·7H 2 O, 0.01g / L CaCl 2 2H 2 O, 0.001g / L FeSO 4 ·7H 2 O, 1....
Embodiment 2
[0042] [Example 2] The degradation characteristics of MZ-5 to benzo [a] anthracene
[0043] 1. Degradation characteristics of MZ-5 to benzo[a]anthracene under different pH conditions (4-10)
[0044] MZ-5 was inoculated in fresh inorganic salt culture solution containing 200mg / L benz[a]anthracene, and placed in a constant temperature incubator at 30°C and 140r / min for enrichment until the logarithmic growth phase. Centrifuge the enriched solution at 6000r / min for 6min, discard the supernatant, resuspend with sterilized phosphate buffer and centrifuge again. After repeating the operation three times, adjust the OD600 of the bacterial suspension to 0.2, and refrigerate for later use .
[0045] Use 1mol / L HCl (43mL concentrated hydrochloric acid to 500mL with deionized water to prepare) and 1mol / L NaOH (4gNaOH is dissolved in 100mL deionized water to prepare) to sterilize fresh The pH value of the inorganic salt culture solution was adjusted to pH=4, pH=5, pH=6, pH=7, pH=8, pH=9...
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