Pseudomonas psychrophila HA-4 low-temperature sulfamethoxazole degrading bacteria as well as screening method and application thereof
A technology of sulfamethoxazole and degrading bacteria, applied in the fields of bioengineering and environmental engineering
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Embodiment 1
[0036] The bacterial strain Pseudomonas psychrophila provided by the invention is separated from activated sludge, and is screened at a set temperature (10°C). The specific screening steps are as follows:
[0037] (1) Take 20mL of fresh activated sludge from the aerobic tank of Harbin Taiping Urban Sewage Plant and add 100mL of inorganic salt culture medium containing 20mg / L sulfamethoxazole as the sole carbon and energy source into a triangular flask at a temperature of 10 °C, cultured in a shaker with a rotation speed of 150r / min for one week to obtain the enriched bacterial liquid;
[0038] (2) Take 10mL of the enriched bacteria liquid and add it to a 100mL sulfamethoxazole inorganic salt culture medium conical flask, repeat the culture conditions of step (1) 3 to 4 times, each time the concentration of sulfamethoxazole is graded Improve, finally to 100mg / L, to obtain the domesticated bacterial liquid, and obtain the OD of the domesticated bacterial liquid 660 =1.192;
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Embodiment 2
[0053] The present invention uses sulfamethoxazole as the only carbon and nitrogen source medium to degrade it and strain it Pseudomonas psychrophila The application of HA-4 growth status research, its steps are as follows:
[0054] strain Pseudomonas psychrophilaHA-4 under the conditions of temperature 10°C, initial pH value 6.0, shaker speed 150r / min, 50mL Erlenmeyer flask liquid volume 20mL, the inoculum amount of HA-4 bacterial suspension was 15%, sulfamethoxazole The initial concentration of the degradation system was 100 mg / L, and the culture was continued for 192 hours, and the concentration of sulfamethoxazole was detected every 24 hours. Using high performance liquid chromatography, the chromatographic conditions are Waters e2695 Alliance HPLC, and the chromatographic column is Symmetry -C18 column (4.6×250 mm, 5 μm); the mobile phase is acetonitrile: formic acid water=40:60 (V / V); the detection wavelength is 265nm; the column temperature is 30°C; the flow rate i...
Embodiment 3
[0058] The present invention uses sulfamethoxazole as the only carbon and nitrogen source medium for its degradation research:
[0059] The initial concentration of the sulfamethoxazole degradation system will be changed in Example 2, and the other steps are the same as in Example 2. image 3 It is the degradation situation of HA-4 bacterial suspension to different initial concentration sulfamethoxazole; By image 3 Visible, strain Pseudomonas psychrophila HA-4 has efficient degradation ability to sulfamethoxazole, and can tolerate the concentration of sulfamethoxazole up to 200mg / L.
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