Providencia alcalifaciens and application thereof in degrading tetracycline and generating phytohormone
A Providencia and auxin technology is applied in the field of antibiotic-contaminated microbial remediation to achieve the effect of reducing residues
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Embodiment 1
[0045] Screening, isolation, purification and identification of Example 1 bacterial strain TC1
[0046] 1. Screening method for a tetracycline-degrading bacterial strain TC1
[0047] 1. Material preparation
[0048] Soil source for strain screening: It was taken from fertilized vegetable fields of farmhouses in Zuolong Village, Chaling County, Zhuzhou City, Hunan Province. The soil samples were sealed with sampling bags, and brought back to the laboratory at 4°C for storage for later use.
[0049] LB medium: peptone 10.0g, yeast extract powder 5.0g, NaCl 10.0g, pH7.0-7.2, distilled water to 1L; solid medium plus 18.0g agar powder. Sterilize at 121°C for 15 minutes.
[0050] Inorganic salt medium (MSM medium): 5mL phosphate buffer solution (KH 2 PO 4 8.5g·L -1 、K 2 HPO 4 ·H 2 O2 1.75g·L -1 、Na 2 HPO 4 12H 2 O 33.4g·L -1 , NH 4 Cl 5.0g L -1 ), 3.0mL 22.5g·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...
Embodiment 2
[0074] Degradation of Tetracycline by Example 2 Bacterial Strain TC1 in Inorganic Salt Medium Containing Tetracycline and Peptone
[0075] Streak the strain TC1 and culture it on the LB plate 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 a TC content of 10 mg·L -1 containing 10g·L -1 Cultured in tryptone-based MSM medium, and samples were taken after 3, 5, and 9 d (days); no addition of strain TC952 was used as a control (CK). Sample liquid at 10000r·min -1 After centrifugation for 1 min, the supernatant was filtered with a 0.22 μm filter membrane, and the residual amount of TC was measured by HPLC after filtration (tetracycline determination method was the same as in Example 1).
[0076] The results showed that the control TC without adding bacteria was hydrolyzed, and after adding bacteria TC1, the degradation of TC was significantly accel...
Embodiment 3
[0077] Example 3 Qualitative Test of Auxin Secretion of Bacterial Strain TC1
[0078] Inoculate the strain TC1 drawn on the LB solid medium into the LB liquid medium, shake and culture at 125-150 rpm, 28-30°C for 18-24 hours, and then inoculate it into the King culture with 2% inoculation volume (V / V) In the basal medium (TC1 treatment group), 2% sterile water was added to the King's medium for treatment as a negative blank control (CK-), placed at 125 rpm, and shaken at 28° C. for 48 hours (each treatment was repeated three times). At room temperature, pipette 50ul of the treatment solution of each group onto the white color comparison plate, and add 50μL of the color comparison solution at the same time; observe the color change of each group after dark treatment for 15 minutes, if the color is pink, the reaction is positive, indicating that it can secrete and produce IAA, and the darker the color, the stronger the ability to secrete IAA; otherwise, it is negative ( Figure...
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