A strain of Pseudomonas taiwanensis capable of degrading tetracycline and its application
A Taiwan Pseudomonas and tetracycline technology, applied in the field of microbial remediation of organic pollutants, can solve problems such as environmental microbial diversity and ecological security threats
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Embodiment 1
[0025] Example 1 Screening, separation, purification and identification of strain TC952
[0026] 1. Screening method of tetracycline-degrading strain TC952
[0027] 1. Material preparation
[0028] Sludge source for strain screening: Taken from the farmhouse fertilized vegetable field in Zuolong Village, Chaling County, Zhuzhou City, Hunan Province.
[0029] LB medium: 10.0 g of peptone, 5.0 g of yeast extract powder, 10.0 g of NaCl, pH 7.0-7.2, dilute to 1 L with distilled water, and add 18.0 g of agar powder to the solid medium. Sterilize at 121°C for 15min.
[0030] Mineral salt medium (MSM): 5 mL of phosphate buffered solution (KH 2 PO 4 8.5g·L -1 , K 2 HPO 4 ·H 2 O21.75g·L -1 , Na 2 HPO 4 ·12H 2 O 33.4g·L -1 , NH 4 Cl 5.0g·L -1 ), 3.0mL22.5g·L -1 MgSO 4 solution (MgSO 4 ·7H 2 O 46.125g·L -1 ), 1.0mL 0.25g·L -1 FeCl 3 solution (FeCl 3 ·6H 2 O 0.42g·L -1 ), 1.0mL 36.4g·L -1 CaCl 2 Solution (CaCl 2 ·2H 2 O 48.22 g·L -1 ), 1.0mL trace element so...
Embodiment 2
[0054] The determination of phosphorus-dissolving ability of embodiment 2 strain TC952
[0055]Qualitative test of the phosphorus-dissolving function of the strain: inoculate the strain TC952 into LB medium by streaking method, culture in a 30°C incubator for 24 hours, then pick a single colony and inoculate it on the PKO solid medium, and set up 3 repeated experiments , placed in a 37°C incubator for 5-7 days, observe whether there is a transparent circle formed by the hydrolysis of dissolved phosphorus around the colony in the medium, and observe the size of the transparent circle, measure the diameter of the hydrolysis circle and the colony circle with a ruler, and take pictures . According to the ratio of the diameter of the hydrolysis circle to the colony circle, it is judged whether the strain has phosphorus-dissolving effect and the relative size of the phosphorus-dissolving ability. Among them, PKO medium formula: glucose 10g L -1 , (NH 4 ) 2 SO 4 0.5g·L -1 , NaC...
Embodiment 3
[0057] Example 3 Degradation of Tetracycline by Bacterial Strain TC952 in Inorganic Salt Medium Containing Tetracycline and Peptone
[0058] Streak the strain TC952 on the LB plate and culture it at 30°C for 20 hours, pick a single colony and inoculate it in the LB liquid medium, 30°C, 150r min -1 After culturing in a shaker for 20 hours, inoculate at a ratio of 2% (v / v) to TC content of 5, 10, 20, 50 mg·L -1 containing 10g·L -1 Cultured in tryptone-based MSM medium, and samples were taken after 6 d (days); the strain TC952 was not added as a control (CK). After the solution was filtered, the residual amount of TC was measured by HPLC (tetracycline determination method was the same as that in Example 1). The results showed that after 6 days, the control tetracycline without adding bacteria was hydrolyzed, and after adding bacteria TC952, the degradation of tetracycline was significantly accelerated and strengthened. The hydrolysis rate was 32.70%, 33.18%, 23.26%, and 20.32%...
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