A kind of aniline degrading bacteria and its application
A kind of aniline-degrading bacteria and aniline technology, applied in the direction of bacteria, biological water/sewage treatment, microorganisms, etc., can solve the problems of low-concentration aniline wastewater advanced treatment and zero discharge, high cost, high equipment requirements, etc., to achieve safe use, Good growth, no secondary pollution effect
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Embodiment 1
[0031] Embodiment 1, separation and purification of bacterial classification
[0032] Wash the printing and dyeing sludge, take 20g of sludge and put it in 200mL inorganic salt liquid culture medium with aniline (50mg / L) as the only carbon and nitrogen source for 7 days, then transfer to aniline concentration of 10% inoculum 100mg / L inorganic salt liquid medium for acclimatization, and after culturing for 7 days, transfer to aniline concentration of 150mg / L inorganic salt liquid medium with 10% inoculum size for acclimatization. Take the culture solution and dilute it in a solid medium with aniline as the only carbon and nitrogen source. After culturing for 7 days, pick a single colony and streak it for purification. , select one of the dominant strains AN-4a.
[0033] Wherein, the formula (1L) of inorganic salt liquid medium is: NaCl 0.2g, K 2 HPO 4 0.2g, KH 2 PO 4 0.2g, trace element solution 2mL.
[0034] Recipe of trace element solution (1L): FeSO 4 300mg, CuSO 4 ...
Embodiment 2
[0036] Embodiment 2, the physiological and biochemical test identification of bacterial strain
[0037] Preliminary physiological and biochemical identification of the screened strain AN-4a, including colony morphology detection, Gram determination, anaerobic growth test, oxidase test, contact enzyme reaction, lysine decarboxylase test, ornithine decarboxylase test , arginine dihydrolase experiment, etc., said test method adopts the routine test method of measuring this kind of physiological and biochemical indicators in the art.
[0038] The results showed that the strain AN-4a was Gram-negative aerobic bacteria, rod-shaped, white, round, smooth and opaque, slightly protruding, with neat edges. Oxidase positive, catalase positive, lysine decarboxylase negative, ornithine decarboxylase negative, arginine dihydrolase negative. The growth temperature is 15-45°C, and the growth pH is 5-10.
Embodiment 3
[0039] Embodiment 3, the 16S rDNA identification of bacterial strain
[0040] The strain AN-4a was identified as Pigmentiphaga daeguensis sp. by 16S rRNA sequence analysis and Biolog microbial identification system identification. Specific steps are as follows:
[0041] The DNA of strain AN-4a was extracted by conventional phenol method, and the 16S rDNA sequence was amplified with universal primers.
[0042] The PCR reaction system (50 μL) was: 5 μL of 10X PCR buffer, 4 μL of dNTP, 1 μL of each primer, 2.5 μL of DNA template, 0.25 μL of TakaraTaq enzyme, and 36 μL of ultrapure water. The PCR amplification program was 94°C for 3min; 94°C for 1min, 52°C for 1min, 72°C for 1.5min, 30 cycles; 72°C for 10min. The amplified product was detected by agarose electrophoresis, and the results were shown in figure 1 . The amplified products were sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing. The sequencing result is shown in SEQ ID NO:1.
[0043] The measured 16S r...
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